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Effect of Cleistanthin a on Vascular Tension In EFFECT OF CLEISTANTHIN A ON VASCULAR TENSION IN ISOLATED GOAT ARTERY A dissertation submitted in partial fulfilment of the requirements for the degree of Doctor of Medicine in Physiology (Branch V) of the Tamil Nadu Dr. M. G. R. Medical University, Chennai – 600 032 Department of Physiology Christian Medical College Vellore May 2020 Department of Physiology Phone: +91 416 228 4268 Christian Medical College Fax: +91 416 226 2788, Bagayam 226 2268 Vellore 632002 Email: Tamil Nadu [email protected] CERTIFICATE This is to certify that this dissertation work titled Effect of Cleistanthin A on vascular tension in isolated goat artery is a bona fide, original work of the candidate Dr.Srisangeetha G with registration number 201715354, for the partial fulfillment of the rules and regulations for the Doctor of Medicine (M.D.) in the branch of Physiology (Branch V) examination to be held in May 2020 by the Tamil Nadu Dr. MGR Medical University, Chennai. This thesis has not been submitted, in part or full, to any other university. Dr. Elizabeth Tharion Dr. Sathya Subramani Professor & Head Professor & Guide Department of Physiology Department of Physiology Christian Medical College Christian Medical College Vellore – 632002 Vellore – 632002 i DECLARATION I hereby declare that the investigations that form the subject matter of this dissertation titled Effect of Cleistanthin A on vascular tension in isolated goat artery, was carried out by me during my term as a post graduate student in the Department of Physiology, Christian Medical College, Vellore. This thesis has not been submitted in part or full to any other university. Dr. Srisangeetha G Department of Physiology Christian Medical College Vellore – 632002 ii Department of Physiology Phone: +91 416 228 4268 Christian Medical College Fax: +91 416 226 2788, Bagayam 226 2268 Vellore 632002 Email: Tamil Nadu [email protected] CERTIFICATE This is to certify that this dissertation work titled Effect of Cleistanthin A on vascular tension in isolated goat artery is a bona fide, original work of the candidate Dr. Srisangeetha G with registration number 201715354, for the partial fulfillment of the rules and regulations for the Doctor of Medicine (M.D.) in the branch of Physiology (Branch V) examination to be held in May 2020 by the Tamil Nadu Dr. MGR Medical University, Chennai. This thesis has not been submitted, in part or full, to any other university. Dr. Anna B Pulimood Principal Christian Medical College Vellore – 632002 iii Department of Physiology Phone: +91 416 228 4268 Christian Medical College Fax: +91 416 226 2788, Bagayam 226 2268 Vellore 632002 Email: Tamil Nadu [email protected] CERTIFICATE This is to certify that this dissertation work titled Effect of Cleistanthin A on vascular tension in isolated goat artery of the candidate Dr. Srisangeetha G with registration number 201715354 for the award of Doctor of Medicine (M.D.) in the branch of Physiology (Branch V). I personally verified the urkund.com website for the purpose of plagiarism check. I found that the uploaded thesis file from abstract to conclusion shows thirteen percent plagiarism in the dissertation. Dr. Sathya Subramani Professor & Guide Department of Physiology Christian Medical College Vellore – 632002 iv ANTI-PLAGIARISM REPORT The above result is a screenshot of the report generated by the Plagiarism Web Tool on Urkund.com used as per the guidelines of the university. v ACKNOWLEDGEMENT The work presented here is the sum of inspiration, advice and critique derived from several individuals and their vast and deep experiences. I sincerely thank, ✓ Dr. Sathya Subramani mam, my guide and mentor, for her invaluable advice, guidance and encouragement throughout the study. ✓ Dr. Soosai Manickam sir, for his priceless help and support. ✓ Dr. Renu mam, for her advice and encouragement. ✓ Dr. Solomon Satishkumar sir for his advice and support. ✓ Dr. Silviya Rajakumari mam, Dr. Anand Bhaskar sir, Dr. Vinay Timothy Oomen sir for their support and motivation. ✓ Dr. Elizabeth mam, Dr. Neetu mam, Dr. Upasana mam and Dr. Anandit for their kind support and encouragement. ✓ My friends, aravindhan, kawin, sajo, alen, elanchezian for their wishes and cooperation throughout my study. My juniors tephilla, shikha, deena, roshini, Sneha for their valuable support. ✓ Geetha ma’am, for taking care of all things administrative. Her support made running research projects painless and efficient. Natarajan sir, Selvam sir, and Vijayanand, nalinamma for keeping the laboratories supplied and ready. CMC Fluid Research Grant Committee, for funding the study. I am greatly indebted to my parents, my husband, my in laws and my sister for all their love and support throughout the study. My special thank to my vi love mithruthan for understanding and tolerating me. Above all I thank God Almighty for giving me strength and support all the time. vii Table of Contents ABSTRACT .................................................................................................... 1 1. INTRODUCTION ...................................................................................... 3 2. AIMS & OBJECTIVES .............................................................................. 7 2.1. Aim ....................................................................................................... 7 2.2. Objectives ............................................................................................. 7 3. REVIEW OF LITERATURE ..................................................................... 8 3.1. Toxicology ........................................................................................... 8 3.2. Method of suicide in india ................................................................... 9 3.3. Cleistanthus collinus: ......................................................................... 10 3.4. Cleistanthus collinus-Botanical profile .............................................. 12 3.5. Cleistanthus collinus – plant morphology.......................................... 13 3.6. Cleistanthus collinus- Benificial effects ............................................ 14 3.7. Cleistanthus collinus- Chemical profile ............................................. 14 3.8. Cleistanthus collinus – Modes of consumption ................................. 16 3.9. Cleistanthus collinus- Clinical profile ............................................... 16 3.10 Cleistanthus collinus poisoning – Clinical presentation(15–17) ...... 17 3.11. Cleistanthus collinus poisoning - Treatment(17–19) ....................... 19 3.12 Cleistanthus collinus poisoning – Mortality rate .............................. 19 3.13. Cleistanthus collinus poisoning – Mechanism of action ................. 20 viii 3.14. Cleistanthus collinus - Various studies ........................................... 21 3.15. Chromatography: ............................................................................. 23 3.16. Liquid –liquid partition chromatography (LLPC): .......................... 24 3.17. High-performance liquid chromatography (HPLC): ....................... 24 3.18 Small arteries are resistant vessels: ................................................... 24 3.19. Vascular smooth muscle : ................................................................ 25 3.20. Vascular adrenoceptors: ................................................................... 25 3.21. Alpha 1 adrenoceptor subtypes: ....................................................... 26 3.22. Alpha 1 adrenoceptor subtypes: ....................................................... 26 3.23. Ion channel in blood vessel and vascular tone: ................................ 27 3.24. Isolated tissue preparation to study the functions of vascular smooth muscle: ...................................................................................................... 30 3.25. The rationale behind the study: ........................................................ 34 4. MATERIALS AND METHODS .............................................................. 35 5.STATISTICAL ANALYSIS ..................................................................... 58 6. RESULTS ................................................................................................. 62 7. DISCUSSION ........................................................................................... 76 8. CONCLUSION ......................................................................................... 78 9. BIBLIOGRAPHY ..................................................................................... 79 10.ANNEXURE……………………………………………………………86 ix ABSTRACT BACKGROUND Suicidal poisoning with Cleistanthus collinus was most common in rural south India. The consumption of leaves was either in the form of boiled decoction(fresh leaves boiled) or fresh leaves blended and filtered. Death rate was about 37.5% and was stated to be very high with the boiled decoction. Death occurs due to vasoplegic shock. There was no promising antidote to date. In this study, we isolated one of the two toxic compounds Cleistanthin A from aqueous extract of dried leaves of Oduvan. The effect of Cleistanthin A on vascular tension in the isolated goat artery was then studied. AIM To observe the effect of Cleistanthin A isolated by preparatory HPLC on vascular tension in circular and longitudinal strips of small arteries from goat legs. METHOD Shade dried leaves of C.collinus were soaked for 24hrs and boiled for 15 min. After cooling, this extract (DLRTB) was subjected to
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