Diabetes mellitus is one of the most public metabolic disorders. It is mainly classified into type 1 and type 2. Echinochrome (Ech) is a pigment from sea urchins that has antioxidant, anti-microbial, anti-inflammatory and chelating abilities. The present study aimed to investigate the anti-diabetic mechanisms of Ech pigment in
streptozotocin-induced diabetic rats. Thirty-six male Wistar albino rats were divided into two main groups (18 rats/group). Each group was divided into 3 subgroups (6 rats/subgroup); control, diabetic and Ech subgroups.
Diabetic models were induced by a single dose of streptozotocin (60 mg/kg, i.p) for type 1 diabetes and by a high fat diet for 4 weeks before the injection of streptozotocin (30 mg/kg, i.p) for type 2 diabetes. Diabetic groups were treated orally with Ech (1 mg/kg body weight in 10% DMSO) daily for 4 weeks. Ech groups showed a reduction in the concentrations of glucose, globulins, triglycerides (TG), total cholesterol (TC), low density lipoprotein cholesterol (LDL-C), creatinine, urea, uric acid, malondialdehyde (MDA) and the activities of
arginase, aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP) and gamma-glutamyltransferase (GGT).
While, it caused general increase in the levels of insulin, total bilirubin (TB), direct bilirubin (DB), indirect bilirubin (IB), total protein (TP), albumin, nitric oxide (NO) and the activities of glucose-6-phosphate dehydrogenase (G6PD), hexokinase, glutathione-S-transferase (GST), superoxide dismutase (SOD) and glutathione reduced (GSH). The histopathological investigation showed partial restoration of pancreatic islet cells and clear improvement in the hepatic and kidney architecture.
The results of this study clearly show that Ech has anti-diabetic potential in both types of diabetes. The possible anti-diabetic mechanisms of Ech involving improved glucose metabolism, restoration of β cells, improve insulin secretion, improve insulin signaling and antioxidant activity.
List of contents
I. Introduction
Aim of work
II. Materials and methods
II.1. Chemicals and reagents
II.2. Sea urchin Collection
II.3. Echinochrome (Ech) extraction
II.4. Experimental animals
II.5. Ethical Consideration
II.6. Induction of type 1 diabetes mellitus (T1DM)
II.7. Induction of type 2 diabetes mellitus (T2DM)
II.8. Experimental design
II.9. Determination of the physical parameters
II.9.1. Body weight
II.9.2. Urine volume
II.9.3. Hot plate test
II.9.4. Wire suspension
II.10. Animal handling and specimen collection
II.11. Samples preparation
II.11.1. Serum preparation
II.11.2. Liver and kidney homogenate preparation
II.11.3. Histopathological examination
II.12. Biochemical assessment
II.12.1. Diabetic markers
II.12.1.1. Determination of glucose
II.12.1.2. Determination of Insulin
II.12.1.3. Determination of arginase
II.12.1.4. Determination of Hexokinase (HK)
II.12.1.5. Determination of glucose-6-phosphate dehydrogenase (G6PDH)
II.12. 2. Serum biomarkers for liver function
II.12.2.1. Determination of serum aminotransferase enzymes (ASAT, ALAT)
II.12.2.2. Determination of alkaline phosphatase
II.12.2.3. Determination of gamma-glutamyltransferase (GGT)
II.12.2.4. Determination of total bilirubin, direct &indirect
II.12.2.5. Determination of total protein
II.12.2.6. Determination of serum albumin and globulins
II.12. 3. Determination of lipid profile
II.12.3.1. Determination of serum triglycerides (TG)
II.12.3.2. Determination of serum total cholesterol (TC)
II.12.3.3. Determination low density lipoprotein cholesterol (LDL-C)
II.12.3.4. Determination of High density lipoprotein cholesterol (HDL-C)
II.12. 4. Determination of kidney Function tests
II.12.4.1. Determination of creatinine and creatinine clearance
II.12.4.2. Determination of uric acid
II.12.4.3. Determination of urea
II.12. 5. Determination of Oxidative Stress parameters
II.12.5.1. Determination of lipid peroxide (Malandialdehyde)
II.12.5.2. Determination of glutathione reduced (GSH)
II.12.5.3. Determination of catalase (CAT)
II.12.5.4. Determination of superoxide dismutase (SOD)
II.12.5.5. Determination of glutathione-S-transferase (GST)
II.12.5.6. Determination of nitric oxide (NO)
II.13. Histological examination
II.14. Statistical analysis
III. Results
III.1. Physical parameters
III.1.1. Body weight
III.1.2. Urine volume
III.1.3. Hot plate test
III.1.4. Wire suspension
III.2. Diabetic markers
III.2.1. Serum glucose
III.2.2. Insulin
III.2.3. Serum arginase
III.2.4. Liver hexokinase
III.2.5. Liver glucose-6-phosphate dehydrogenase (G6PD)
III.3. Serum biomarkers for liver function
III.3.1. Serum aspartate aminotransferase (AST)
III.3.2. Serum alanine aminotransferase (ALT)
III.3.3. Serum alkaline phosphatase (ALP)
III.3.4. Serum gamma glutamyl transferase (GGT)
III.3.5. Serum total bilirubin (TB)
III.3.6. Serum direct bilirubin (DB)
III.3.7. Serum indirect bilirubin (IB)
III.3.8. Serum total protein (TP)
III.3.9. Serum albumin
III.3.10. Serum globulins
III.3.11. Albumin/globulins ratio (A/G)
III.4. Lipid profile
III.4.1. Serum triglycerides (TG)
III.4.2. Serum total cholesterol (TC)
III.4.3. Serum low density lipoprotein cholesterol (LDL-C)
III.4.4. Serum high density lipoprotein cholesterol (HDL-C)
III.5. Kidney Function tests
III.5.1. Serum creatinine
III.5.2. Urine creatinine
III.5.3. Creatinine clearance
III.5.4. Serum uric acid
III.5.5. Urine uric acid
III.5.6. Serum urea
III.5.7. Urine urea
III.6. Oxidative stress parameters in liver
III.6.1. Liver malondialdehyde (MDA)
III.6.2. Liver glutathione reduced (GSH)
III.6.3. Liver catalase (CAT)
III.6.4. Liver superoxide dismutase (SOD)
III.6.5. Liver glutathione-S-transferase (GST)
III.6.6. Liver nitric oxide (NO)
III.7. Oxidative stress parameters in kidney
III.7.1. Kidney malondialdehyde (MDA)
III.7.2. Kidney glutathione reduced (GSH)
III.7.3. Kidney catalase (CAT)
III.7.4. Kidney superoxide dismutase (SOD)
III.7.5. Kidney glutathione-S-transferase (GST)
III.7.6. Kidney nitric oxide (NO)
III.8. Histopathological examination
III.8.1. Histopathological examination of pancreas
III.8.2. Histopathological examination of liver
III.8.3. Histopathological examination of kidney
IV. Discussion
Conclusion
V. Summary
VI. References
Research Objectives and Thematic Focus
The primary research objective of this thesis is to evaluate the antidiabetic efficacy and potential therapeutic mechanisms of the Echinochrome pigment, extracted from the sea urchin species Paracentrotus lividus, in streptozotocin-induced type 1 and type 2 diabetic rat models.
- Extraction and preparation of the Echinochrome pigment from sea urchin shells and spines.
- Assessment of the pigment's impact on key metabolic, liver function, kidney function, and lipid profile markers.
- Evaluation of the in vivo antioxidant potential of Echinochrome by measuring oxidative stress parameters in liver and kidney tissues.
- Histopathological analysis of the structural changes in the pancreas, liver, and kidneys of diabetic rats treated with Echinochrome.
- Investigation of the potential antidiabetic mechanisms, including improvements in insulin secretion, glucose metabolism, and antioxidant defense.
Excerpt from the Book
II.3.Echinochrome (Ech) extraction
Pigments in the shells and spines were isolated by the Amarowicz method with slight modifications (Amarowicz et al., 1994; Kuwahara et al., 2009). After removal of the internal organs, the shells and spines were washed with a stream of cold water, air-dried at 4°C for 2 days in the dark and then were grounded. The powders (5 g) were dissolved by gradually adding 10 ml of 6 M HCl. The pigments in the solution were extracted 3 times with the same volume of diethyl ether. The ether layer collected was washed with 5% NaCl until the acid was almost removed. The ether solution including the pigments was dried over anhydrous sodium sulfate and the solvent was evaporated under reduced pressure. The extract including the polyhydroxylated naphthoquinone pigment was stored at -30°C in the dark.
Summary of Chapters
I. Introduction: Provides a comprehensive overview of glucose metabolism, the pathogenesis of diabetes mellitus, the role of oxidative stress in diabetic complications, and the potential of marine-derived bioactive compounds such as Echinochrome.
II. Materials and methods: Details the experimental procedures including chemical preparation, sea urchin collection, animal induction of diabetes, and the various biochemical and histopathological techniques used for analysis.
III. Results: Presents the experimental findings regarding physical parameters, diabetic markers, liver and kidney function tests, oxidative stress markers, and histopathological observations across different study groups.
IV. Discussion: Interprets the findings by comparing them with existing scientific literature, detailing the mechanisms behind the antidiabetic and antioxidant effects observed in the study.
V. Summary: Lists the key findings and conclusions observed regarding the impact of Echinochrome on diabetic rat models.
Keywords
Diabetes, Echinochrome, Oxidative stress, Pancreas, Liver, Kidney, Histopathology, Streptozotocin, Glucose metabolism, Insulin secretion, Lipid profile, Hepatotoxicity, Nephrotoxicity, Antioxidant activity, Paracentrotus lividus.
Frequently Asked Questions
What is the core subject of this research thesis?
The research explores the potential of the pigment Echinochrome, derived from sea urchins, as a therapeutic agent for managing metabolic disorders, specifically type 1 and type 2 diabetes.
What are the primary focus areas of this study?
The study focuses on the metabolic, antioxidant, and histopathological effects of Echinochrome in diabetic rat models, evaluating its ability to normalize blood glucose, improve organ function, and reduce oxidative stress.
What is the central research question?
The study aims to determine if Echinochrome can effectively act as an antidiabetic agent and to identify the biological mechanisms through which it restores metabolic homeostasis in diabetic subjects.
Which scientific methodology was utilized?
The research utilized streptozotocin-induced diabetic rat models for both type 1 and type 2 diabetes. It employed a combination of biochemical assays (for blood and tissue analysis) and histopathological examinations (microscopic analysis of organs) to assess the curative potential of the pigment.
What topics are covered in the main body?
The main body covers the extraction of the pigment, the experimental design and induction of diabetes, the rigorous measurement of physiological and biochemical markers, and detailed histological investigations of affected tissues.
Which keywords characterize this work?
Key terms include Diabetes, Echinochrome, Oxidative stress, Pancreas, Liver, Kidney, Histopathology, and Antioxidant activity.
How was type 2 diabetes specifically induced in the study?
Type 2 diabetes was induced by feeding the rats a high-fat diet for 4 weeks, followed by a single intraperitoneal injection of streptozotocin (30 mg/kg).
What impact does Echinochrome have on pancreatic cells?
The research concludes that Echinochrome facilitates a partial restoration of pancreatic islet cells and reduces beta-cell damage, which contributes to improved insulin secretion and better glycemic control.
What significant changes were observed in liver and kidney markers?
Treated subjects showed a reduction in markers of cellular damage and oxidative stress (such as MDA, AST, ALT, and GGT) and an increase in antioxidant enzymes (such as GSH, SOD, and CAT), indicating restored organ architecture and improved detoxification functions.
- Quote paper
- M.Sc Ayman Mohamed (Author), 2018, The Metabolic Effects of Echinochrome Pigment Extracted from Sea Urchin on Diabetic Rats, Munich, GRIN Verlag, https://www.grin.com/document/387206
