| This study is based on TCM syndrome differentiation type 2 diabetes experience and early type 2 diabetic rats model of disease and syndrome research. Experiment to choose the high fat and sugar to feed 5 weeks after intraperitoneal injection of disposable chain of small dose of Streptozotozin (STZ) method can induce T2DM rat model. In type 2 diabetic rats model of 5-6 weeks after the successful stage belongs to Qi and yin deficiency, phlegm and blood stasis syndrome. This stage can study the rat illness physiology and the characteristics of TCM syndrome, which focus on from the perspective of pancreas damage and protection, to explore the diagnosis and treatment qi, phlegm and remove blood stasis of TCM Yin square model rat islet function and mechanism.The thesis is divided into two parts:literature review and experimental research. Literature review includes the progress in prevention and treatment of traditional Chinese medicine in type 2 diabetes and research progress of protective effect of islet beta cell in type 2diabetes. Experiment includes the type 2 diabetic rats model replication of the study of Chinese and western medicine, with Supplementing qi and nourishing yin and activating blood and dissipating phlegm treatment effect and mechanism of type 2 diabetic rats.Methods:Experiment chooses 55 healthy male SD rats, weight 190±10 g, and adaptive feed 1 w. According to the weight were randomly divided into normal group and 10 and made module only 45. Normal group rats given basic feed, feed made module according to the high-fat high-calorie 1%plus small dose of STZ intraperitoneal injection of 30 mg/kg method to establish the rat model with T2DM.Sixth weeks of screening out of 30 consistent with the standard model (fasting blood glucose greater than or equal to 11.1mmol/L) in rats, and randomly selected rats were divided into model group 10, western medicine group 10, traditional Chinese medicine intervention group was 10.The rats of control group were given normal diet, while the other group was given high fat and high calorie diet. Fed to 11 week, traditional Chinese medicine group were treated with Supplementing qi and nourishing yin and activating blood and dissipating phlegm (4.80g/kg/d) by gavage, western medicine treatment group were given pioglitazone hydrochloride (1.35mg/kg/d) intragastric administration. The model group and the normal group were fed with distilled water. The experimental rats were continuous lavage 2w. Changes were observed and recorded the rats hair color, behavior, diet, stool and urine etc. Changes of the rat’s hair color, behavior, diet, stool and urine volume were observed and recorded Weighing 1 times a week. Electrogeneratedchemiluminescence method for the determination of serum insulin; fasting blood glucose; insulin action index (IAI); Biochemical method of determination of serum triglycerides (TG), high-density lipoprotein (HDL-c), serum low density lipoprotein (LDL-c), FFA, pancreatic tissue iNOS; Enzyme-linked immunoassay detection serum 6-ketone prostaglandin F1 alpha, serum TXB2, serum AngⅡ, serum PI, islet amylin, islet VECF, islet HGF, islet JAK-2,islet PKB. Pancreatic tissue pathological slices use HE staining, aldehyde compound red dye, D-PAS staining and Mallory staining to observe islet.Results:(1) The conditions of the model rats:①Model group rats are characterized by cluster, listlessness, greasy hair, tail easily wet and ecchymosis. Compared with the normal group, the rats in the model group, the amount of dietary water intake, urine volume increased significantly, decreased body weight (P＜0.01). ②The sugar and lipid metabolism:Compared with normal group, model group rats FPG, Fins, a significant rise in IRI, IAI significantly lower, (P＜0.01). Compared with the normal group, the model group rats serum TG, LDH-C and FFA increased significantly, HDL-c decreased (P＜0.05 or P< 0.01).③The islet function: Compared with the normal group, model group rats serum PI, insulin amylin levels increased significantly (P＜0.01).④The blood and blood vessels, regulating factor:Compared with the normal group, the serum 6-keto-PGF1a of rats in model group was significantly decreased, content of TXB2, TXB2/6-keto-PGF1a, Ang Ⅱ increased significantly (P＜0.05 or P＜0.01). ⑤The pancreas:Compared with the normal group, the model group rats islets increased within iNOS, reduce VEGF, HGF, JAK2, the content of PKB (P＜0.05orP＜0.01). Microscopically islet obvious atrophy showed irregular shape, structure disorder, islet cell cavity deformation, beta cell area significantly decreased, microvascular basement membrane thickening in the islet, islet collagen fibers increased obviously. (2) Supplementing qi and nourishing yin and activating blood and dissipating phlegm on the therapeutic effect of model rats:① Chinese traditional medicine group rats characterized by increased activity, responsive, smooth hair, tail no damp and dramatically reduce petechiae. Compared with the normal group, the rats in the model group, the amount of dietary water intake, urine volume increased significantly, decreased body weight (P＜0.01). Compared with model group, western medicine group rats drinking and reduced the volume of urine, weight gain (P＜9.01), the diet is no significant difference (P>0.05).The Chinese medicine group urine volume had decreased trend (P>0.05), food and water intake was reduced, increased body weight (P＜0.01 or P＜0.05). Compared with the western medicine group, Chinese medicine group rats drinking water decreased (P＜.01), diet, urine volume and body weight did not change significantly (P>0.05). ②The sugar and lipid metabolism:Compared with the model group,Chinese medicine group and Western medicine group rats FPG, Fins, IRI were significantly decreased, IAI increased significantly (P<0.01).Compared with the western medicine group, Chinese medicine group rats FPG, Fins, IRI, IAI had no change (P>0.05). Compared with the model group, western medicine group and Chinese medicine group serum TG decreased, HDL-c decreased (P＜0.05 or P<0.01), HDL-c increased trend, FFA decreased trend (P>0.05).Compared with the western medicine group, difference between Chinese traditional medicine group rats serum TG, LDH-c, HDL-c and FFA had no statistical significance (P>0.05). ③The islet function:Compared with the model group, western medicine group and Chinese medicine group rats serum PI, insulin amylin content decreased significantly (P< 0.01).Compared with the western medicine group, Chinese medicine group rats serum PI, islets of amylin showed no significant difference (P>0.05).④The blood and blood vessels, regulating factor:Compared with the model group, western medicine group and Chinese medicine group 6-keto-PGF1a rats were significantly increased, decreased the content of TXB2, TXB2/6-keto-PGFla, Ang Ⅱ (P＜0.01).Compared with the western medicine group, Chinese medicine group rats were TXB2,6-keto-PGF1a, TXB2/6-keto-PGF1a, Ang Ⅱ unchanged (P>0.05).⑤The pancreas:Compared with the model group, western medicine group iNOS decreased (P＜0.01), VEGF, HGF, JAK2, PKB content had no change (P>0.05). Microscopically the islet structure is neat, pancreatic islet atrophy, a small amount of cells in vacuoles, beta cells increased, the average area islet in microvascular basement membrane thickening, islet collagen fiber discreaseing. Compared with model group, Chinese medicine group rat pancreas iNOS decreased significantly (P＜0.01) VEGF and HGF, JAK2, PKB significantly higher (P<0.05 or P＜0.01), the VEGF and HGF, JAK2 significant rise in the western medicine group (P<0.01). Microscopically islet structure is clear, has good cell morphology, islet cavitation and significantly reduced, average beta cell area increased significantly, islet microvascular basement membrane thickening is not apparent, islet collagen fiber discreaseing, within the area of islet beta cells increased significantly in the western medicine group (P<0.01)Conclusion:Experiment to choose the high fat and sugar to feed 5 weeks after intraperitoneal injection of disposable chain of small dose of Streptozotozin (STZ) method can induce T2DMrat model. In type 2 diabetic rats model of 5-6 weeks after the successful stage belongs to Qi and yin deficiency, phlegm and blood stasis syndrome. The stage shows not only the characteristics of the TCM and at the same time also has the qi and Yin deficiency and phlegm stasis syndrome. The model not only has the western disease in type 2 diabetes IR characteristic, and vascular regulating disorders, microenvironment abnormality and pancreatic islet beta cell damage. Compound Chinese medicine can not only improve the rat model of type 2 diabetes syndromes, but also can improve the glucolipid metabolic disorder of diabetes model rats and pancreatic islet beta cells and the micro environment of damage. Traditional Chinese medicine compound for the treatment of type 2 diabetes mellitus in addition to sugar, lipid-lowering effect,also involved in regulating vascular active factor, improve the pancreas islet microenvironment and promote restoration of the beta cells Pioglitazone hydrochloride has obviously improve the model rats of sugar lipid metabolic disorder and IR resistant effect, reducing pancreatic fibrosis,but the beta cell damage and induce the microenvironment and without obvious effect. Therefor, the regulating effect of multi compound traditional Chinese medicine has certain advantages in the prevention and treatment of diabetes mellitus. |
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