pathomorphological and biochemical studies on the effect of gymnema sylvestre and chromium in ...

381 Pages·2015·10.18 MB·English

PATHOMORPHOLOGICAL AND BIOCHEMICAL
STUDIES ON THE EFFECT OF GYMNEMA
SYLVESTRE AND CHROMIUM IN INDUCED
DIABETES IN RATS

KAVITHA RANI B

DEPARTMENT OF VETERINARY PATHOLOGY
VETERINARY COLLEGE, HEBBAL, BENGALURU – 560024
KARNATAKA VETERINARY, ANIMAL AND FISHERIES
SCIENCES UNIVERSITY, BIDAR
MAY, 2015
i
PATHOMORPHOLOGICAL AND BIOCHEMICAL
STUDIES ON THE EFFECT OF GYMNEMA
SYLVESTRE AND CHROMIUM IN INDUCED
DIABETES IN RATS

Thesis submitted to the
Karnataka Veterinary, Animal and Fisheries Sciences University, Bidar.
in partial fulfillment of the requirements for the award of the degree of
DOCTOR OF PHILOSOPHY
in
Veterinary Pathology
By
KAVITHA RANI B

DEPARTMENT OF VETERINARY PATHOLOGY
VETERINARY COLLEGE, HEBBAL, BENGALURU – 560024
KARNATAKA VETERINARY, ANIMAL AND FISHERIES
SCIENCES UNIVERSITY, BIDAR
MAY, 2015
ii
KARNATAKA VETERINARY, ANIMAL AND FISHERIES
SCIENCES UNIVERSITY, BIDAR.
DEPARTMENT OF VETERINARY PATHOLOGY
VETERINARY COLLEGE, HEBBAL, BENGALURU-560024
CERTIFICATE
This is to certify that the thesis entitled “PATHOMORPHOLOGICAL AND
BIOCHEMICAL STUDIES ON THE EFFECT OF GYMNEMA SYLVESTRE AND
CHROMIUM IN INDUCED DIABETES IN RATS” submitted by Mrs. KAVITHA RANI B.,
I.D. No. DVHK 1229 in partial fulfillment of the requirements for the award of DOCTOR OF
PHILOSOPHY in Veterinary Pathology of the Karnataka Veterinary, Animal and Fisheries
Sciences University, Bidar is a record of bonafide research work carried out by her during the
period of her study in this University under my guidance and supervision and the thesis has not
previously formed the basis for the award of any degree, diploma, associateship, fellowship or
other similar titles.
Date: 25.05.2015
Place: Bengaluru Prof. SUGUNA RAO
(Major Advisor)
Approved by :
Major Advisor : ________________________________
(Prof. SUGUNA RAO)
Nominated External Examiner : ________________________________
(Prof. G.A. BALASUBRAMANIAM)
Members : 1. ________________________________
(Prof. M.L. SATYANARAYANA)
2. ________________________________
(Prof. H.D. NARAYANASWAMY)
3. ________________________________
(Dr. S. M. BYREGOWDA)
4. ________________________________
(Dr. C. ANSAR KAMRAN)
5. ________________________________
(Dr. N.B. SHRIDHAR)
iii
Affectionately Dedicated to
My Beloved
Father - Dr. B.S. NAIDU
Mother – Smt. JAYALAKSHMI
My
“GUIDE”
and My
“FAMILY”

iv
ACKNOWLEDGEMENTS
I hereby render my humble salutations to the Almighty God and bow in front
of him for making this feat of doctoral work possible, which is my lifetime
achievement.
“Guru Brahma, Guru Vishnu, Guru Devoh Maheshwarah” … explains the bond
between myself and my mentor guru Prof. Suguna Rao, my Major advisor, who is an
untiring teacher, a facilitator guide, a task oriented advisor with a personal
commitment, a skeptical supervisor and an honest researcher with an unbiased and
open mind. I will always carry the image of you with gratitude, for it is you who has
shown me the path less trodden, chiefly responsible for the outcome of my thesis.
My respects and heartfelt regards are expressed for
Prof. M. L. Satyanarayana, an unassuming person whose kindly facilitation and
timely help made this work possible. He always had words to inspire and nudge me
towards the goal, which I cherish with all respect.
My thanks and regards are for Prof. H. D. Narayanaswamy, who was
always there for moral support and guidance in times of ambiguity throughout the
period of PhD programme at the department.
My sincere gratitude and regards are specially extended to
Dr. S. M. Byregowda, Director (IAH&VB) and my advisory committee member
without whose facilitation for carrying out molecular work, my research outcome
would have been incomplete.
With due respect, I thank Dr. Ansar Kamran, C., an accomplished
veterinary clinician, for giving necessary inputs on technical aspects of the
research work and also critically reviewing the application of the outcome in the
field.
My heartfelt thanks are for Dr. N. B. Shridhar, who was always willing to
help and advice whenever required during the study period. I sincerely thank you
for the technical and professional guidance, and physical facilities extended for
carrying out the research work.
My special thanks and regards are for Dr. K. M. Purushotham, of AINPBT
laboratory of IAH&VB, who taught me the molecular methods of estimation, and
was personally involved in my analytical work, in the last phases of my research.
v
I sincerely acknowledge the facilitation done by Dr. Rajeshwari, Professor,
Department of LPM, Veterinary College, Bangalore for laboratory rats and
accessories.
The most important person, directly involved in the execution of the
experiment, was Gurikar R.Y., whom I will always remember for many things and
especially the times when we worked together.
My senior PhD colleagues namely, Vetrivel and Deepak JN, have helped me
both physically as well as technically, throughout the study period. I thank you
friends for being there in times of need.
My respects and regards are extended to Mrs. Nadira, for facilitating the
material needs for the study, and also for extending moral support during the PhD
programme. I also acknowledge the moral support given by Dr. Vishwanath, and my
junior colleague Dr. Anjan Kumar K. R. of Bangalore.
Sincere thanks are especially due for my colleagues Manjunath, S.S. and
Shambhulingappa Baddi of Shimoga for giving me as much time as possible to
concentrate on PhD programme.
My thanks are also for Mr. Gnanesha of Shimoga, Mrs. Rangalakshmi, and
Late Smt. Lakshmi of Bangalore. I acknowledge Obleshappa for the physical help
during the study period at the sheds, and Nagaraj and Pillaraju of the Department
of Veterinary Pathology, Veterinary College, Bengaluru.
I place my thanks also with Mrs. Kumuda and Mrs. Devamma, of the
AINPBT lab, IAH&VB for making my time happier.
My regards are in order for Mrs. Betageri, Mr. Girish and Mr. Mahesh of
the Library at Veterinary College, Bangalore, for unfolding the wealth of knowledge
both physically and virtually.
I cannot forget my fellow PhD colleagues, B. P. Shivashankar and Avinash
Lakkawar, both of them diverse characters, who made my PhD programme, an
unforgettable time. Thank you.
My regards to my classmate friends Deepti, Sharada, Suresh Patel, Ganga
Naik, Malathi, Rajani, Sunilchandra, Ravikumar, Prakash Choori, Anand,
Kalmath, Sreenivas murthy, Bipin and Manegar for being just …..friends.
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I also thank Satish Mundas, Bhairegowda, Lakshmi, Simitha, Kishore,
Kavya, Ganavi, Deepa of Pathology, Veena (my roommate at Kalpavruksha hostel),
Mahesh Kadagi and Santhosh C. R. of Pharmacology, for their support.
Words fail to express the magnanimous support and constant inspiration
rendered by my ‘beloved father’, Dr B. S. Naidu, an altruistic human, a geneticist
and a researcher himself who acted as a sounding board for my research work, and
my dearest mother, Smt Jayalakshmi which looked after my wonderful children
Vishnupriya and Shantipriya because of which I could concentrate on my work.
Words cannot express my respectful regards for my eldest brother-in-law
Ramesh Kumar and my sister Sukanya who were my guardians and protective like
parents, and Prateek in whose abode, I could peacefully carry out my work at
Bangalore.
I acknowledge the moral support, understanding and co-operation by my dear
husband, Dr. S. Goverdhan, who himself was a student at the Medical college
during this period.
I thank my sister Dr Uma and brother-in-law Gopalkrishna who supported
my parents at home during my absence, and their children Chandini and Meghana.
My special thanks to Mrs. Monisha, Bharat and Aaditya, and their family
for facilitating several essential requirements during the fag end of my research.
I also thank my in-laws in Bangalore, Mrs. Anusuyamma, Mrs. Shashikala
and family and Mrs. Devika and family for their moral support during my PhD
programme.
My thanks for M/s Vinayaka Printers for a neat and wonderful printing job
executed.
I thank the Karnataka Veterinary Animal and Fisheries Sciences University,
Bidar for giving me opportunity to pursue my doctoral studies.
Lastly, I bow to my Alma-mater – Veterinary College, KVAFSU, Bengaluru.

Bengaluru,
May, 2015 KAVITHA RANI, B.

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CONTENTS
CHAPTER TITLE PAGE No.
I INTRODUCTION 1-5
II REVIEW OF LITERATURE 6-73
III MATERIAL AND METHODS 74-112
IV RESULTS 113-218
V DISCUSSION 219-283
VI SUMMARY 284-291
VII BIBLIOGRAPHY 292-330
VIII ABSTRACT 331

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LIST OF TABLES
Table Page
Title
No No
Mean (±SE) animal body weights (g) of normal control, chromium
1 control, diabetic and diabetic treatment groups on 3rd, 15th, 30th and 169
45th day.
Mean (±SE) haemoglobin (g %) values of normal control, chromium
2 control, diabetic and diabetic treatment groups on 3rd, 15th, 30th and 171
45th day.
Mean (±SE) serum glucose (mg/dL) values of normal control,
3 chromium control, diabetic and diabetic treatment groups on 3rd, 15th, 173
30th and 45th day.
Mean (±SE) serum cholesterol (mg/dL) values of normal control,
4 chromium control, diabetic and diabetic treatment groups on 3rd, 15th, 175
30th and 45th day.
Mean (±SE) serum triglyceride (mg/dL) values of normal control,
5 chromium control, diabetic and diabetic treatment groups on 3rd,15th, 177
30th and 45th day.
Mean (±SE) serum very low density lipoprotein (mg/dL) values of
6 normal control, chromium control, diabetic and diabetic treatment 179
groups on 3rd, 15th, 30th and 45th day.
Mean (±SE) serum alanine aminotransferase (ALT) (IU/L) values of
7 normal control, chromium control, diabetic and diabetic treatment 181
groups on 3rd, 15th, 30th and 45th day.
Mean (±SE) serum aspartate aminotransferase (AST) (IU/L) values
8 of normal control, chromium control, diabetic and diabetic treatment 183
groups on 3rd, 15th, 30th and 45th day.
Mean (±SE) liver superoxide dismutase (units / min / mg of protein)
9 values of normal control, chromium control, diabetic and diabetic 185
treatment groups on 15th, 30th and 45th day.
Mean (±SE) liver catalase (nmoles of H O decomposed / min / mg
2 2
10 protein) values of normal control, chromium control, diabetic and 187
diabetic treatment groups on 15th, 30th and 45th day.
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Table Page
Title
No No
Mean (±SE) liver glutathione peroxidase (μ moles of glutathione
utilized / min / mg protein) values of normal control, chromium
11 189
control, diabetic and diabetic treatment groups on 15th, 30th and 45th
day.
Mean (±SE) serum insulin (µU/L) values of normal control,
12 chromium control, diabetic and diabetic treatment groups on 15th, 191
30th and 45th day.
Mean (±SE) insulin positive pancreatic β cells (%) of normal control,
13 chromium control, diabetic and diabetic treatment groups on 15th, 193
30th and 45th day.
Mean (±SE) Insulin gene expression (folds) in pancreas in normal
14 control, chromium control, diabetic and diabetic treatment groups on 195
15th, 30th and 45th day
Mean (±SE) Insulin receptor gene expression (folds) in liver in
15 normal control, chromium control, diabetic and diabetic treatment 197
groups on 15th, 30th and 45th day
Mean (±SE) GLUT-4 gene expression (folds) in liver in normal
16 control, chromium control, diabetic and diabetic treatment groups on 199
15th, 30th and 45th day

x