Journal of Clinical and Diagnostic Research, ISSN - 0973 - 709X

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Dr Mohan Z Mani

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On Sep 2018




Prof. Somashekhar Nimbalkar

"Over the last few years, we have published our research regularly in Journal of Clinical and Diagnostic Research. Having published in more than 20 high impact journals over the last five years including several high impact ones and reviewing articles for even more journals across my fields of interest, we value our published work in JCDR for their high standards in publishing scientific articles. The ease of submission, the rapid reviews in under a month, the high quality of their reviewers and keen attention to the final process of proofs and publication, ensure that there are no mistakes in the final article. We have been asked clarifications on several occasions and have been happy to provide them and it exemplifies the commitment to quality of the team at JCDR."



Prof. Somashekhar Nimbalkar
Head, Department of Pediatrics, Pramukhswami Medical College, Karamsad
Chairman, Research Group, Charutar Arogya Mandal, Karamsad
National Joint Coordinator - Advanced IAP NNF NRP Program
Ex-Member, Governing Body, National Neonatology Forum, New Delhi
Ex-President - National Neonatology Forum Gujarat State Chapter
Department of Pediatrics, Pramukhswami Medical College, Karamsad, Anand, Gujarat.
On Sep 2018




Dr. Kalyani R

"Journal of Clinical and Diagnostic Research is at present a well-known Indian originated scientific journal which started with a humble beginning. I have been associated with this journal since many years. I appreciate the Editor, Dr. Hemant Jain, for his constant effort in bringing up this journal to the present status right from the scratch. The journal is multidisciplinary. It encourages in publishing the scientific articles from postgraduates and also the beginners who start their career. At the same time the journal also caters for the high quality articles from specialty and super-specialty researchers. Hence it provides a platform for the scientist and researchers to publish. The other aspect of it is, the readers get the information regarding the most recent developments in science which can be used for teaching, research, treating patients and to some extent take preventive measures against certain diseases. The journal is contributing immensely to the society at national and international level."



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Professor and Head
Department of Pathology
Sri Devaraj Urs Medical College
Sri Devaraj Urs Academy of Higher Education and Research , Kolar, Karnataka
On Sep 2018




Dr. Saumya Navit

"As a peer-reviewed journal, the Journal of Clinical and Diagnostic Research provides an opportunity to researchers, scientists and budding professionals to explore the developments in the field of medicine and dentistry and their varied specialities, thus extending our view on biological diversities of living species in relation to medicine.
‘Knowledge is treasure of a wise man.’ The free access of this journal provides an immense scope of learning for the both the old and the young in field of medicine and dentistry as well. The multidisciplinary nature of the journal makes it a better platform to absorb all that is being researched and developed. The publication process is systematic and professional. Online submission, publication and peer reviewing makes it a user-friendly journal.
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Dr Saumya Navit
Professor and Head
Department of Pediatric Dentistry
Saraswati Dental College
Lucknow
On Sep 2018




Dr. Arunava Biswas

"My sincere attachment with JCDR as an author as well as reviewer is a learning experience . Their systematic approach in publication of article in various categories is really praiseworthy.
Their prompt and timely response to review's query and the manner in which they have set the reviewing process helps in extracting the best possible scientific writings for publication.
It's a honour and pride to be a part of the JCDR team. My very best wishes to JCDR and hope it will sparkle up above the sky as a high indexed journal in near future."



Dr. Arunava Biswas
MD, DM (Clinical Pharmacology)
Assistant Professor
Department of Pharmacology
Calcutta National Medical College & Hospital , Kolkata




Dr. C.S. Ramesh Babu
" Journal of Clinical and Diagnostic Research (JCDR) is a multi-specialty medical and dental journal publishing high quality research articles in almost all branches of medicine. The quality of printing of figures and tables is excellent and comparable to any International journal. An added advantage is nominal publication charges and monthly issue of the journal and more chances of an article being accepted for publication. Moreover being a multi-specialty journal an article concerning a particular specialty has a wider reach of readers of other related specialties also. As an author and reviewer for several years I find this Journal most suitable and highly recommend this Journal."
Best regards,
C.S. Ramesh Babu,
Associate Professor of Anatomy,
Muzaffarnagar Medical College,
Muzaffarnagar.
On Aug 2018




Dr. Arundhathi. S
"Journal of Clinical and Diagnostic Research (JCDR) is a reputed peer reviewed journal and is constantly involved in publishing high quality research articles related to medicine. Its been a great pleasure to be associated with this esteemed journal as a reviewer and as an author for a couple of years. The editorial board consists of many dedicated and reputed experts as its members and they are doing an appreciable work in guiding budding researchers. JCDR is doing a commendable job in scientific research by promoting excellent quality research & review articles and case reports & series. The reviewers provide appropriate suggestions that improve the quality of articles. I strongly recommend my fraternity to encourage JCDR by contributing their valuable research work in this widely accepted, user friendly journal. I hope my collaboration with JCDR will continue for a long time".



Dr. Arundhathi. S
MBBS, MD (Pathology),
Sanjay Gandhi institute of trauma and orthopedics,
Bengaluru.
On Aug 2018




Dr. Mamta Gupta,
"It gives me great pleasure to be associated with JCDR, since last 2-3 years. Since then I have authored, co-authored and reviewed about 25 articles in JCDR. I thank JCDR for giving me an opportunity to improve my own skills as an author and a reviewer.
It 's a multispecialty journal, publishing high quality articles. It gives a platform to the authors to publish their research work which can be available for everyone across the globe to read. The best thing about JCDR is that the full articles of all medical specialties are available as pdf/html for reading free of cost or without institutional subscription, which is not there for other journals. For those who have problem in writing manuscript or do statistical work, JCDR comes for their rescue.
The journal has a monthly publication and the articles are published quite fast. In time compared to other journals. The on-line first publication is also a great advantage and facility to review one's own articles before going to print. The response to any query and permission if required, is quite fast; this is quite commendable. I have a very good experience about seeking quick permission for quoting a photograph (Fig.) from a JCDR article for my chapter authored in an E book. I never thought it would be so easy. No hassles.
Reviewing articles is no less a pain staking process and requires in depth perception, knowledge about the topic for review. It requires time and concentration, yet I enjoy doing it. The JCDR website especially for the reviewers is quite user friendly. My suggestions for improving the journal is, more strict review process, so that only high quality articles are published. I find a a good number of articles in Obst. Gynae, hence, a new journal for this specialty titled JCDR-OG can be started. May be a bimonthly or quarterly publication to begin with. Only selected articles should find a place in it.
An yearly reward for the best article authored can also incentivize the authors. Though the process of finding the best article will be not be very easy. I do not know how reviewing process can be improved. If an article is being reviewed by two reviewers, then opinion of one can be communicated to the other or the final opinion of the editor can be communicated to the reviewer if requested for. This will help one’s reviewing skills.
My best wishes to Dr. Hemant Jain and all the editorial staff of JCDR for their untiring efforts to bring out this journal. I strongly recommend medical fraternity to publish their valuable research work in this esteemed journal, JCDR".



Dr. Mamta Gupta
Consultant
(Ex HOD Obs &Gynae, Hindu Rao Hospital and associated NDMC Medical College, Delhi)
Aug 2018




Dr. Rajendra Kumar Ghritlaharey

"I wish to thank Dr. Hemant Jain, Editor-in-Chief Journal of Clinical and Diagnostic Research (JCDR), for asking me to write up few words.
Writing is the representation of language in a textual medium i e; into the words and sentences on paper. Quality medical manuscript writing in particular, demands not only a high-quality research, but also requires accurate and concise communication of findings and conclusions, with adherence to particular journal guidelines. In medical field whether working in teaching, private, or in corporate institution, everyone wants to excel in his / her own field and get recognised by making manuscripts publication.


Authors are the souls of any journal, and deserve much respect. To publish a journal manuscripts are needed from authors. Authors have a great responsibility for producing facts of their work in terms of number and results truthfully and an individual honesty is expected from authors in this regards. Both ways its true "No authors-No manuscripts-No journals" and "No journals–No manuscripts–No authors". Reviewing a manuscript is also a very responsible and important task of any peer-reviewed journal and to be taken seriously. It needs knowledge on the subject, sincerity, honesty and determination. Although the process of reviewing a manuscript is a time consuming task butit is expected to give one's best remarks within the time frame of the journal.
Salient features of the JCDR: It is a biomedical, multidisciplinary (including all medical and dental specialities), e-journal, with wide scope and extensive author support. At the same time, a free text of manuscript is available in HTML and PDF format. There is fast growing authorship and readership with JCDR as this can be judged by the number of articles published in it i e; in Feb 2007 of its first issue, it contained 5 articles only, and now in its recent volume published in April 2011, it contained 67 manuscripts. This e-journal is fulfilling the commitments and objectives sincerely, (as stated by Editor-in-chief in his preface to first edition) i e; to encourage physicians through the internet, especially from the developing countries who witness a spectrum of disease and acquire a wealth of knowledge to publish their experiences to benefit the medical community in patients care. I also feel that many of us have work of substance, newer ideas, adequate clinical materials but poor in medical writing and hesitation to submit the work and need help. JCDR provides authors help in this regards.
Timely publication of journal: Publication of manuscripts and bringing out the issue in time is one of the positive aspects of JCDR and is possible with strong support team in terms of peer reviewers, proof reading, language check, computer operators, etc. This is one of the great reasons for authors to submit their work with JCDR. Another best part of JCDR is "Online first Publications" facilities available for the authors. This facility not only provides the prompt publications of the manuscripts but at the same time also early availability of the manuscripts for the readers.
Indexation and online availability: Indexation transforms the journal in some sense from its local ownership to the worldwide professional community and to the public.JCDR is indexed with Embase & EMbiology, Google Scholar, Index Copernicus, Chemical Abstracts Service, Journal seek Database, Indian Science Abstracts, to name few of them. Manuscriptspublished in JCDR are available on major search engines ie; google, yahoo, msn.
In the era of fast growing newer technologies, and in computer and internet friendly environment the manuscripts preparation, submission, review, revision, etc and all can be done and checked with a click from all corer of the world, at any time. Of course there is always a scope for improvement in every field and none is perfect. To progress, one needs to identify the areas of one's weakness and to strengthen them.
It is well said that "happy beginning is half done" and it fits perfectly with JCDR. It has grown considerably and I feel it has already grown up from its infancy to adolescence, achieving the status of standard online e-journal form Indian continent since its inception in Feb 2007. This had been made possible due to the efforts and the hard work put in it. The way the JCDR is improving with every new volume, with good quality original manuscripts, makes it a quality journal for readers. I must thank and congratulate Dr Hemant Jain, Editor-in-Chief JCDR and his team for their sincere efforts, dedication, and determination for making JCDR a fast growing journal.
Every one of us: authors, reviewers, editors, and publisher are responsible for enhancing the stature of the journal. I wish for a great success for JCDR."



Thanking you
With sincere regards
Dr. Rajendra Kumar Ghritlaharey, M.S., M. Ch., FAIS
Associate Professor,
Department of Paediatric Surgery, Gandhi Medical College & Associated
Kamla Nehru & Hamidia Hospitals Bhopal, Madhya Pradesh 462 001 (India)
E-mail: drrajendrak1@rediffmail.com
On May 11,2011




Dr. Shankar P.R.

"On looking back through my Gmail archives after being requested by the journal to write a short editorial about my experiences of publishing with the Journal of Clinical and Diagnostic Research (JCDR), I came across an e-mail from Dr. Hemant Jain, Editor, in March 2007, which introduced the new electronic journal. The main features of the journal which were outlined in the e-mail were extensive author support, cash rewards, the peer review process, and other salient features of the journal.
Over a span of over four years, we (I and my colleagues) have published around 25 articles in the journal. In this editorial, I plan to briefly discuss my experiences of publishing with JCDR and the strengths of the journal and to finally address the areas for improvement.
My experiences of publishing with JCDR: Overall, my experiences of publishing withJCDR have been positive. The best point about the journal is that it responds to queries from the author. This may seem to be simple and not too much to ask for, but unfortunately, many journals in the subcontinent and from many developing countries do not respond or they respond with a long delay to the queries from the authors 1. The reasons could be many, including lack of optimal secretarial and other support. Another problem with many journals is the slowness of the review process. Editorial processing and peer review can take anywhere between a year to two years with some journals. Also, some journals do not keep the contributors informed about the progress of the review process. Due to the long review process, the articles can lose their relevance and topicality. A major benefit with JCDR is the timeliness and promptness of its response. In Dr Jain's e-mail which was sent to me in 2007, before the introduction of the Pre-publishing system, he had stated that he had received my submission and that he would get back to me within seven days and he did!
Most of the manuscripts are published within 3 to 4 months of their submission if they are found to be suitable after the review process. JCDR is published bimonthly and the accepted articles were usually published in the next issue. Recently, due to the increased volume of the submissions, the review process has become slower and it ?? Section can take from 4 to 6 months for the articles to be reviewed. The journal has an extensive author support system and it has recently introduced a paid expedited review process. The journal also mentions the average time for processing the manuscript under different submission systems - regular submission and expedited review.
Strengths of the journal: The journal has an online first facility in which the accepted manuscripts may be published on the website before being included in a regular issue of the journal. This cuts down the time between their acceptance and the publication. The journal is indexed in many databases, though not in PubMed. The editorial board should now take steps to index the journal in PubMed. The journal has a system of notifying readers through e-mail when a new issue is released. Also, the articles are available in both the HTML and the PDF formats. I especially like the new and colorful page format of the journal. Also, the access statistics of the articles are available. The prepublication and the manuscript tracking system are also helpful for the authors.
Areas for improvement: In certain cases, I felt that the peer review process of the manuscripts was not up to international standards and that it should be strengthened. Also, the number of manuscripts in an issue is high and it may be difficult for readers to go through all of them. The journal can consider tightening of the peer review process and increasing the quality standards for the acceptance of the manuscripts. I faced occasional problems with the online manuscript submission (Pre-publishing) system, which have to be addressed.
Overall, the publishing process with JCDR has been smooth, quick and relatively hassle free and I can recommend other authors to consider the journal as an outlet for their work."



Dr. P. Ravi Shankar
KIST Medical College, P.O. Box 14142, Kathmandu, Nepal.
E-mail: ravi.dr.shankar@gmail.com
On April 2011
Anuradha

Dear team JCDR, I would like to thank you for the very professional and polite service provided by everyone at JCDR. While i have been in the field of writing and editing for sometime, this has been my first attempt in publishing a scientific paper.Thank you for hand-holding me through the process.


Dr. Anuradha
E-mail: anuradha2nittur@gmail.com
On Jan 2020

Important Notice

Original article / research
Year : 2023 | Month : February | Volume : 17 | Issue : 2 | Page : CC01 - CC04 Full Version

Dehydroepiandrosterone and Acute Stress Attenuation: An Interventional Rodent Study


Published: February 1, 2023 | DOI: https://doi.org/10.7860/JCDR/2023/61231.17535
Tamilselvan Kuppusamy, Gayathri Veeraraghavan, Santhi Silambanan, Muraliswaran Perumal, Padmavathi Ramaswamy

1. Professor and Head, Department of Physiology, Sri Venkateshwaraa Medical College Hospital and Research Centre, Ariyur, Puducherry, India. 2. Head, Department of Test Facility Management, Centre for Toxicology and Developmental Research, Sri Ramachandra Institute for Higher Education and Research, Chennai, Tamil Nadu, India. 3. Professor, Department of Biochemistry, Sri Ramachandra Institute for Higher Education and Research, Chennai, Tamil Nadu, India. 4. Professor and Head, Department of Biochemistry, Sri Venkateshwaraa Medical College Hospital and Research Centre, Puducherry, India. 5. Professor, Department of Physiology, Sri Ramachandra Institute for Higher Education and Research, Chennai, Tamil Nadu, India.

Correspondence Address :
Padmavathi Ramaswamy,
Professor, Department of Physiology, Sri Ramachandra Medical College and Research Institute, Sri Ramachandra Institute for Higher Education and Research, Porur, Chennai-600116, Tamil Nadu, India.
E-mail: rpadmavathi@sriramachandra.edu.in

Abstract

Introduction: Stress activates hypothalamo-pituitary-adrenal axis leading to the release of glucocorticoid that mediates the stress response. This adaptive response is self-limited but if persistent for prolonged periods can lead to disease states. Nature has endowed the body with efficient buffer systems to attenuate the stress effects and Dehydroepiandrosterone (DHEA), a steroid hormone with neuromodulatory functions is implicated as an efficient candidate to buffer stress.

Aim: To assess the effect of prophylactic administration of DHEA in the attenuation of acute stress in male Wistar rats.

Materials and Methods: This interventional study was carried out at centre for Toxicology and Developmental Research, Sri Ramachandra Institute of Higher Education and Research, Chennai, between June 2021 and August 2021, in compliance with the animal welfare guidelines of CPCSEA, and in accordance to the protocol approved by Institutional Animal Ethics Committee. The 18 male Wistar rats approved for the study were segregated into 3 groups with 6 animals in control (no stress) group, 6 in stress group and 6 in intervention group that received DHEA prophylactically 30 min before stress procedure. Animals in stress and intervention groups were subjected to one hour immobilisation stress. Blood samples were collected from all animals after the stress period and serum corticosterone, the stress marker, was estimated. The data were expressed as mean±standard error of mean (mean±SEM) and Mann-Whitney U test was used to test the significant difference between the: (i) control and stress groups; (ii) stress and study groups; and (iii) control and study groups. The p-value <0.05 was considered significant. The analysis was done using SPSS version 23.0.

Results: The values of corticosterone in control, stress and intervention groups were 26.6±4.4 ng/mL, 51.6±3.9 ng/mL and 23.4±3.6 ng/mL, respectively. Significant difference in the mean serum corticosterone levels with p-value 0.013 between control and stress groups and with p-value 0.008 between stress and DHEA groups were observed.

Conclusion: It could be observed from the findings that prophylactic DHEA administration attenuated acute stress efficiently in male Wistar rats as reflected by the significant decrease in serum corticosterone levels in the group that received DHEA intervention, thus inferring the efficiency of DHEA in stress buffering.

Keywords

Adrenal androgen, Anxiety, Immobilisation, Psychological stress, Serum corticosterone

The maintenance of internal balance (homeostasis) at times of real or perceived stress is made possible by the evolution of stress response that brings about adaptive changes through the activation of autoregulated neural and hormonal systems. Of the two mechanisms, the hormonal mechanism serves as the key regulator in mediating the Hypothalamic-Pituitary-Adrenal (HPA) axis. By the activation of HPA axis glucocorticoid is ultimately released from adrenal cortex and mediates the adaptive response to stress (1). Glucocorticoids are primarily concerned with ensuring energy availability by mobilisation and distribution of energy to various organ systems in appropriation to demands, both at basal and stressed states thus rendering adaptive response at times of stress an efficient mechanism to meet the demands of the dire moments (2). Glucocorticoids exert action via both non genomic and genomic mechanisms and its effects could be observed almost on all tissues in the body as its receptors are widely distributed in all body systems (3),(4).

These neuroendocrine mediated metabolic, physiological and behavioural adaptations that ensures improved survival in the face of temporary stressors by establishing stability through changes, is referred as allostasis (5),(6). Such adaptations if persistent for prolonged periods or repeated can often lead to physiologic dysregulation with poor health outcomes like increased susceptibility to infections, cardiovascular disease, metabolic syndrome, obesity, cancer, and mental health disorders that constitutes the allostatic load (1).

Stress affects mental health and on the long-term impairs the quality of life. Therefore, in the recent past many research endeavours in the field of mental health have revealed various endogenous biomolecules to possess efficient stress buffering capacity. In this vein, DHEA and its sulphated derivative DHEA-Sulfate (DHEAS) which were previously considered to serve only as a precursor of potent androgen and oestrogens on peripheral conversion, were recently found to have a plethora of beneficial effects that includes stress attenuation as it has antiglucocorticoid properties (7). DHEA is observed to modulate endothelial function, improve insulin sensitivity, reduce inflammation, improve blood flow, regulate body composition, bone metabolism, sexual function, enhance neuroprotection, improve cognition and memory as well counteract the immunosuppressive actions of corticosteroid (8),(9). Age related metabolic derangement, cognitive decline, neurodegenerative disorders as well as the aetiopathology of many psychiatric illness lie with derangement in endogenous DHEA levels (10),(11),(12).

The DHEA, as an antistress agent, is observed to prevent the stress induced inhibition in body weight gain, increase in adrenal weight, and concentration of glucocorticoid receptor levels in liver, thymus, and spleen when administered to stress male Sprague-Dawley rats and it also reduces the lipid peroxidation levels in the liver and heart (13). Similarly, DHEA is also observed to inhibit the stress induced corticosterone mediated inhibition of testosterone in experimentally stressed adult Sprague-Dawley rats (14). Previous research reported a contradictory observation where the administration of sulphated form of DHEA in high-anxiety rats induced anxiolysis while it induced anxiogenesis in low-anxiety rats which infers a stress modulatory effect of DHEA (15).

Though the above mentioned observations have highlighted the efficiency of DHEA in stress buffering, there is paucity of experimental evidence to establish the beneficial effect of prophylactic DHEA administration in attenuation of acute stress. As rodents share physiologic and genetic similarities with human and the stress response could be better studied with appropriate stress paradigm in regulated and controlled conditions from stress induction to interventional outcomes in animal model (16), this study was planned as a preclinical study to assess the beneficial effect of prophylactic DHEA administration in the attenuation of acute stress in Wistar male rats.

Material and Methods

This interventional study was carried out at Centre for Toxicology and Developmental Research, Sri Ramachandra Institute of Higher Education and Research, Chennai, Tamil Nadu, India between July and September 2021. The study was performed in compliance with the animal welfare guidelines published by CPCSEA (17), Government of India and standard operating procedures of CEFTE in accordance to the Protocol approved by Institutional Animal Ethics Committee (IAEC number: IAEC59/SRIHER/659/2019).

Study Procedure

Animal procurement, acclimatisation and housing: Eighteen adult male Wistar rats, as sanctioned by IAEC, were procured from in-vivo Bioscience, Hyderabad. The animals were 60-70 days of age with 200-250 g bodyweight and were acclimatised under laboratory conditions for about 14 days. The animals were housed in rooms maintained at 19-23°C temperature and humidity of 30-70%, with air exchange in the range of 12-15 air changes per hour, respectively. The animals were provided with photoperiod of 12 hour artificial light and 12 hour darkness and were segregated into three groups by simple randomisation technique, viz., six animals in control (non stressed) group, six animals in stress group and six animals in intervention group that received DHEA prophylactically 30 minutes before immobilisation stress. Animals were segregated by the random number generated using Microsoft excel. The animals were housed in groups in polypropylene cags covered with stainless steel grid. Dedusted and autoclaved paddy husk was used as bedding material which was changed on alternate days. The animals were habituated to handling during the acclimatisation period and the same personnel handled the animals on all days and even for the experimental procedures. Animals were provided with standard rodent pelleted feed (Krishna Valley Agrotech LLP, Maharashtra) and portable UV treated water was provided ad libitum in autoclaved bottles.

Treatment: The experiment commenced on the fourth day following the completion of acclimatisation period. The intervention group was administered with intraperitoneal DHEA-at a dose of 25 mg/kg diluted in 2-3 drops of Dimethyl Sulfoxide (DMSO) and then with physiological saline, 30 minutes before the stress procedure (18). DHEA was procured from Avanti Polar Lipids, Alabaster, Alabama.

Acute immobilisation stress: Acute immobilisation stress was inculcated to the stress and intervention groups by immobilisation of the animal in a restrainer (25×7 cm plastic) and adjusting it with plaster tape on the outside so that animal was unable to move. For breathing, 1 cm hole at one end was provided (19). The stress was carried out for a period of one hour between 10 and 11 am. During immobilisation, the rats were not provided with feed and water.

Sample collection and storage: At the end of one hour of immobilisation, blood samples were collected from all three groups: controls (non stressed), stressed, and intervention (stressed 30 min after prophylactic administration of DHEA) groups. Blood samples were collected through retro-orbital plexus under sterile aseptic precautions and allowed to clot in non heparinised tubes. The sera were collected from the samples by centrifugation at 3000 rpm for 15 minutes at room temperature. The serum thus separated was stored in ID tagged eppendorf at -80°C until hormone analysis was performed.

Animal rehabilitation: After sample collection, the animals were returned to animal facility for reuse.

Serum corticosterone estimation: The hormone concentrations were determined as per the instructions provided in 96-well Enzyme Linked Immunosorbent Assay (ELISA) kit for rat corticosterone, procured from Elabscience. The standards were diluted as per the instruction provided in the kit 0-25 ng/mL. The samples were diluted at a ratio of 1:2 and then at 1:4, in duplicates, as the former dilution yielded the response beyond the standard range. Standard and samples were taken in the respective wells and incubated with biotinylated detection antibody followed by HRP conjugate working solution at 37°C for specific period and washed. Plates were treated with substrate and the reaction was terminated using stop solution. The plate absorbance was detected at 450 nm using a microplate reader. Hormone assay was performed by a third party completely blinded from the experiment, as per the kit insert. All data received were included in the results. However, no blinding was done during result assessment.

Statistical Analysis

The data were expressed as mean±standard error of mean (mean±SEM) and Mann Whitney U test was used to test the significant difference between the: (i) control and stress groups; (ii) stress and intervention groups; and (iii) control and intervention groups. Comparison between the three groups was done using Kruskal Wallis test. The p-value <0.05 was considered significant. The analysis was done using SPSS version 23.0.

Results

A total of 18 adult male Wistar rats were included in the study. Six animals were in control group (unstressed), 6 in stress group and 6 in intervention group where animals received DHEA prophylactically 30 minutes before immobilisation stress procedure.

Comparison of the body weight of the animals in the 3 groups showed no significant difference (p=0.271) (Table/Fig 1).

Comparison of serum corticosterone levels between the three groups as calculated by Kruskal Wallis test revealed a significant mean difference existing between the three groups with a p-value 0.007* (Table/Fig 2).

Serum corticosterone level was significantly elevated in stress group when compared to control with p-value 0.013* and the same was attenuated significantly in intervention group with a p-value 0.008* when compared to stress group. No significant difference was observed in the comparison between control and intervention groups (Table/Fig 3).

Discussion

Stress begins as an acute event inducing an adaptive response in the biological system by the release of glucocorticoid into circulation that helps tackle the demands of the moment of crisis. This acute adaptive phenomenon is beneficial and has autoregulatory self-limiting mechanisms. But the persistence of this response for longer periods renders the autoregulatory processes ineffective and ends in disease states. Moreover, stress outcomes are determined by the individual’s vulnerability and the sequels of morbidity are more pronounced among the stress susceptible traits (20). As personality influences anticipatory stress vulnerability and effectiveness of coping interventions, prophylactic measures to attenuate the stress response in this vulnerable population can promise them with a better quality of life. Hence, this study was designed as a preclinical research to evaluate the efficiency of prophylactic DHEA administration in the attenuation of acute stress.

In the present study, it was observed that one hour immobilisation induced stress significantly in the stress group animals which is evident from the significant elevation of serum corticosterone levels in them with a p-value 0.013* when compared to the control group. With prophylactic administration of DHEA the reduction in the stress marker was significant as observed from the comparison between the stress and intervention (stressed with prophylactic administration of DHEA) groups with a p-value 0.008*. On comparing control group values with intervention group, no significant difference was observed reflecting the prophylactic administration of DHEA had significantly reduced serum corticosterone comparable to that of control values observed in non stressed control group. Significance was observed with intergroup difference (p-value 0.007*).

During stress, at the stimulation of Adrenocorticotropic Hormone (ACTH), the biosynthesis of adrenal DHEA begins with the rate-limiting step of importing cholesterol from cytosol to the inner mitochondrial membrane by steroidogenic acute regulatory protein, StAR, which is then converted to pregnenolone by the mitochondrial cholesterol side-chain cleavage enzyme, cytochrome P450scc. Pregnenolone by the action of 17-alpha hydroxylase gets converted to 17OH-pregnenolone (17OH-Preg) and then to DHEA subsequently by the action of 17,20-lyase (21).

Though it was reported that the rodent’s adrenal CYP17A1 lack 17,20-lyase activity, thus rendering the adrenal cortex of adult rodents with compromised capacity to synthesise adrenal androgens, the diurnal rhythm of DHEA and corticosterone metabolites are found to follow a similar temporal pattern in them thus inferring the common source of both steroids from adrenal gland (22),(23). It was also reported that DHEA that circulates in lower concentrations in rodent blood (around 1-3 nM or less) is synthesised from non adrenal steroidogenic source such as gonads, and placenta as well by the nervous system, as evident from the observations of their levels in the blood of adrenalectomised and castrated rats (24).

The DHEA thus formed, is reversibly converted to its sulfate ester-DHEA-S by the enzyme sulfotransferase (SULT2A1) in which form it is predominately found in circulation (about 300 times higher than that of free DHEA, in human). Diurnal variation is exhibited by DHEA but not by DHEA-S, with former having a short terminal half-life while DHEA-S has a much longer half-life thus making the inference that DHEA-S primarily serves as a reservoir of DHEA with a lesser role in physiological functions (25).

The DHEA thus circulating is found to exert its effect on a wide range of tissues like liver, kidney, adipose tissue, reproductive tissues, and central nervous system as well on various neural structures. The hormone DHEA and its sulphated form, DHEA-S, exert their physiological effects either by its direct actions on DHEA-specific plasma membrane receptors coupled via G-protein or through various neuroreceptors like N-Methyl-D-Aspartate (NMDA), aminobutyric-acid-type A, and sigma-1 (S1R) receptors; or by binding to androgen and oestrogen receptors (ARs, ERα, or ERβ) either directly or by their more potent steroid metabolites-testosterone, dihydrotestosterone, and estradiol (26). But further exploration is needed to establish the exact mechanism.

DHEA being a potent antistress hormone is found to exert an inhibitory modulation on stress axis. Budziszewska B et al., in their in-vitro study with Neuro-2A cells observed that neither DHEA nor DHEA-S is found to modify CRH gene promoter activity, implying that DHEA has no modulatory effect on the activation of HPA axis (27). But Taguchi T et al., observed that at supraphysiological doses the hormone is observed to inhibit POMC transcription in-vitro in ACTH secreting cell line AtT-20 and thus decrease the expression of ACTH (28).

Chang LL et al., reported that DHEA attenuated ACTH induced corticosterone release in in-vitro study with rat zona glomerulosa fasciculate cell cultures not only by decreasing the activities of the enzymes involved in the keysteps of corticosterone biosynthesis viz., P450scc, 3β-HSD, 21-hydroxylase and 11β-hydroxylase but also the prime step of ACTH induced cAMP formation. It is observed that the incubation of rat adrenal cells with the substrates of the steroidogenic enzymes viz 25-OH-cholesterol, pregnenolone, progesterone and deoxycorticosterone, respectively corticosterone release was enhanced. But the same was attenuated significantly when incubated in presence of DHEA thus making the inference that DHEA appears to decrease the activities of 3β-HSD, 21-hydroxylase and 11β-hydroxylase. No significant attenuation of actions was observed with P450scc as the binding of 25-OH-cholesterol and P450scc is not affected by DHEA, while only the quantity of P450scc involved in the process is reduced. DHEA inhibits 11β-hydroxylase activity by the mechanism of competitive inhibition and interferes with the formation of the binding complex of 11β-hydroxylase and deoxycorticosterone. DHEA is also observed to decrease the expression of StAR protein that transfers cholesterol from cytoplasmic pool to the inner mitochondrial membrane, by decreasing the translation of its protein (29).

Apostolova G et al., observed that DHEA exerted its antiglucocorticoid effect either by downregulating 11β-HSD1-dependent glucocorticoid regeneration from 11 dehydrocorticosterone, the inactive metabolite, or by enhancing 11β-HSD2-dependent glucocorticoid inactivation into the inactive metabolite in the peripheral tissues. In 2005, they reported that DHEA induced the downregulation of 11β-HSD1-dependent glucocorticoid regeneration in liver, adipose tissue, and kidneys of C57BL/6J mice (30). The same team in 2008 reported their observation that DHEA induced the activity of 11β-HSD2 in a rat cortical collecting duct cell line and in kidneys of C57BL/6J mice and Sprague-Dawley rats whereby the corticosterone was converted to dehydrocorticosterone. DHEA is also reported to attenuate ischaemia/reperfusion-induced oxidative stress in rodent kidney (31).

From the observations of the above mentioned in-vitro studies (Taguchi T et al., 2006, Chang LL et al., 2003, Apostolova G et al., 2005, Apostolova G et al., 2008) it could be inferred that DHEA exerts its antistress effects by inhibiting the transcription of ACTH as well the enzymes involved in adrenal glucocorticoid synthesis (28),(29),(30). It is also observed to enhance the reduction in the plasma concentration of corticosterone by upregulating 11β-HSD2 dependent glucocorticoid inactivation in the kidney and downregulating 11β-HSD1 dependent glucocorticoid regeneration in liver, adipose tissue, and kidneys [28-31]. In accord with these observations, it was observed in the present study that intervention with prophylactic administration of DHEA, 30 min prior to immobilisation stress led to a significant decrease in serum corticosterone levels in the intervention group when compared to the stress group. The attenuation in the glucocorticoid levels was comparable with control values. This observation could be interpreted as the prophylactically administered DHEA had exerted a significant attenuating effect on stress axis and thus the stress response could have been decreased by the above mentioned mechanisms and the corticosterone levels in intervention group was comparable with that of control values, reflecting the efficient role of prophylactically administered DHEA in the attenuation of acute stress.

Limitation(s)

The observations of the present study have made a significant inference on the stress-attenuating potency of DHEA though the molecular mechanism of the stress attenuation by DHEA was not explored. With further molecular research endeavours the exact mechanism of DHEA in stress attenuation could be established.

Conclusion

DHEA, the antistress adrenocorticoid, when administered prophylactically in Wistar rats, is found to significantly attenuate stress at supraphysiological doses. Hence, it could be concluded that DHEA is an efficient antiglucocorticoid and is found to be an effective attenuator of stress in animal model. Further research efforts are recommended to establish its clinical utility.

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DOI and Others

DOI: 10.7860/JCDR/2023/61231.17535

Date of Submission: Nov 03, 2022
Date of Peer Review: Dec 12, 2022
Date of Acceptance: Jan 18, 2023
Date of Publishing: Feb 01, 2023

AUTHOR DECLARATION:
• Financial or Other Competing Interests: None
• Was Ethics Committee Approval obtained for this study? Yes
• Was informed consent obtained from the subjects involved in the study? NA
• For any images presented appropriate consent has been obtained from the subjects. NA

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