| Background:Diabetes mellitus is a metabolic disease characterized by chronic high blood glucose caused by many reasons,mainly caused by defect of insulin secretion and (or) function. At present,the prevalence rate, incidence and the patient population of diabetes rise sharply worldwide. The course of the diabetes is longer, with many complications,and it can cause disability or even death. So it often brings huge economic burden to the patients and the society, and damage the patient’s quality of life seriously. The western medicine has many side effects while the acupuncture treatment of diabetes has curative effect without side effects. Yishu(EX-B3)and Pishu(BL20) are the common acupoints used in clinic to treat diabetes, and they are all in the scope of spinal segments of the pancreas. This experiment observed wether the Yishu(EX-B3)and Pishu(BL20) have different curative effects, and try to explore their effective mechanism.Objectives:(1)The research is to test the effects of Yishu(EX-B3)and Pishu(BL20) on type2diabetic rat about the body weight, blood glucose, serum lipid, insulin, morphology of islet ceells,and to observe wether the Yishu and Pishu have different curative effects on type2diabetes, even though they are both in the scope of spinal segments of the pancreas.(2)The research is to test the effects of Yishu(EX-B3)and Pishu(BL20) on type2diabetic rat about the PDX-1protein and PDX-1, GLUT2,GCK mRNA expression, to explore their effective mechanism.Methods:Research about the curative effect observation of the Yishu (EX-B3) and Pishu (BL20) which are both in the same spinal segments with the pancreas on the T2DM rats:60Male Wistar rats were divided into2groups randomly:normal group(containing8rats) and type2diabetes mell itus model building group ("model build ing group" for short, containing52rats). Model building group was fed with high sugar and high fat fodder for4weeks and then inject streptozotocin(STZ) in the abdominal cavity with35mg/kg. BW to build T2DM model. The successful models were divided into4groups according to their blood glucose:model group(containing10rats), glimepiride gavage group (containing10rats), electro-acupuncture on Yishu group (containing11rats), electro-acupuncture on Pishu group (containing10rats). Gl imepiride gavage group and the electro-acupuncture groups were intervened once a day, five times per week, and continued for4weeks. Observed and recorded the general condition of the rats, and tested the body weight and the blood glucose every weekend. After4weeks,suicided the rats with decollation, and collected the blood, then collected blood serum by centrifuging. Measured the level of serum triglyceride(TG), total cholesterol (TC), low-density lipoprotein cholesterol(LDL), high-density lipoprotein cholesterol (HDL)and glycosylated serum protein (GSP) with biochemical process;measured the level of insulin with radioimmunoassay. Take a part of the pancreatic tissue,and then fixed it with paraformaldehyde, which would be used to observe the islet morphology with HE staining.Research about the mechanism of the Yishu(EX-B3)and Pishu(BL20) which are both in the same spinal segments with the pancreas on the T2DM rats:50Male Wistar rats were divided into2groups randomly:normal group(containing8rats)and type2diabetes mellitus model building group("model building group" for short, containing42rats). Model building group was fed with high sugar and high fat fodder for4weeks and then inject streptozotocin(STZ) in the abdominal cavity with35mg/kg. BW to build T2DM model. The successful models were divided into3groups according to their blood glucose:model group(containing10rats), electro-acupuncture on Yishu group (containing11rats) and electro-acupuncture on Pishu group (containing10rats). The electro-acupuncture groups were intervened once a day, five times per week, and continued for4weeks. After4weeks, suicided the rats with decollation, take a part of the pancreatic tissue, and one part was fixed with paraformaldehyde, which would be used to test the PDX-1protein expression through immunohistochemical method. While another part was fixed with liquid nitrogen, and was used to test the PDX-1. GLUT2, GCK mRNA expression by Real-time PCR method.Results:1.Research about the curative effect observation of the Yishu (EX-B3)and Pishu(BL20) which are both in the same spinal segments with the pancreas on the T2DM rats:(1)Body weight:During the intervention of4weeks, the body weight of the normal group increased gradually. The body weight of the model group decreased gradually, but had no obvious difference compared with that of itself before intervention; While compared with the body weight of the normal group at the same time, there were obvious differences(P<0.05). It was in line with the characteristics of diabetic rats. The body weight of glimepiride gavage group, electro-acupuncture on Yishu group and electro-acupuncture on Pishu group decreased obviously compared with that before intervention, but there was no significant difference compared with that of the model group at the same time.(2)Blood glucose:During the intervention of4weeks, the blood glucose level of normal group maintained around5mmo1/L. While the blood glucose level of model group maintained around20mmo1/L, and had no significant changes during the4weeks. During the4weeks, the blood glucose level of glimepiride gavage group had no significant differences compared with that of the model group at the same time or compared with that of itself before intervention. When compared with the that of the model group at the same time, blood glucose level of electro-acupuncture on Yishu group had no significant differences at the first to third weekends,but dropped obviously at the fourth weekend(P<0.05). When compared with that of itself before intervention, the blood glucose level of electro-acupuncture on Yishu group rose not obviously at the first to second weekends,dropped not obviously at the third weekend and dropped obviously at the fourth weekend(P<0.05). When compared with that of the model group at the same time, the blood glucose level of electro-acupuncture on Pishu group had no significant differences at the first to third weekends,but dropped obviously at the fourth weekend(P<0.05). When compared with that of itself before intervention, the blood glucose level in different stages of electro-acupuncture on Pishu group all dropped, and it dropped obviously at the first and third weekends (P<0.05). When compared with that of itself at last weekend, the blood glucose level dropped at the first and third weekends,rose at the second and fourth weekends. That is to say, the blood glucose level of electro-acupuncture on Pishu group fluctuated obviously.(3)Serum lipid:Compared with the normal group, the TC, HDL, LDL levels of model group rose significantly(P<0.05). Compared with the model group,the TG, TC and LDL levels of the glimepiride gavage group dropped without statistically significant difference; while the HDL level dropped significantly(P<0.05). The TG, LDL and HDL levels of the electro-acupuncture on Yishu group dropped significantly(P<0.05); while the TC level dropped without statistically significant difference. The TC level of the electro-acupuncture on Pishu group dropped significantly(P<0.05); while the TC, LDL and HDL levels dropped without statistically significant difference. Compared with the electro-acupuncture on Yishu group, the TG, TC, LDL and HDL levels of the glimepiride gavage group and the electro-acupuncture on Pishu group had no statistically significant differences.(4)Fast insulin:Compared with the normal group,the fast insulin level of the model group had no statistically significant difference. Compared with the model group, the fast insulin levels of the glimepiride gavage group and the electro-acupuncture on Pishu group dropped signif icantly(P<0.05), while the fast insulin level of the electro-acupuncture on Yishu group had no statistically significant difference with it. Comparison between any two groups among the three treatment groups had no statistically significant difference.(5) Insulin sensitive index (ISI):Compared with the normal group, the ISI of the model group and the three treatment groups dropped significantly(P<0.01). Compared with the model group, the ISI of the glimepiride gavage group,the electro-acupuncture on Yishu group and the electro-acupuncture on Pishu group rose significantly(P<0.05). Compared with the electro-acupuncture on Yishu group, the ISI of the glimepiride gavage group dropped significantly(P<0.05).(6)Morphology of islet cells:Compared with the normal group, the islet of the model group damaged obviously with atrophied islet, irregular edge,cells hypertrophy and differ sizes, cytoplasm light dye and visible vacuolated cells. Compared with the model group, the islet of the glimepiride gavage group and the electro-acupuncture on Yishu group is better, the boundary of the islet is clear,but still with irregular edge, significant proliferation of edge cells, intensive cell nucleus and celsl of differ sizes. The islet of the electro-acupuncture on Pishu group is better than that of the model group, but is worse than the other two treatment groups, with serious damage, irregular forms of cells and exocrine gland bubble stretched into the islet.2. Research about the mechanism of the Yishu (EX-B3) and Pishu (BL20) which are both in the same spinal segments with the pancreas on the T2DM rats:(1)PDX-1protein:Compared with the normal group, the PDX-1protein expression of the model group, the electro-acupuncture on Yishu group and the electro-acupuncture on Pishu group dropped but without statistically significant difference. Compared with the model group,the PDX-1protein expression of the electro-acupuncture on Yishu group and the electro-acupuncture on Pishu group rose but without statistically significant difference. Comparison between the two treatment groups had no statistically significant difference.(2)PDX-1mRNA:Compared with the normal group, the PDX-1mRNA expression of the model group, the electro-acupuncture on Yishu group and the electro-acupuncture on Pishu group dropped significantly (P<0.05). Compared with the model group, the PDX-1mRNA expression of the electro-acupuncture on Yishu group and the electro-acupuncture on Pishu group rose significantly(P<0.05). Comparison between the two treatment groups had no statistically significant difference.(3)GLUT2and GCK mRNA:Compared with the normal group, the GLUT2and GCK mRNA expression of the model group, the electro-acupuncture on Yishu group and the electro-acupuncture on Pishu group dropped significantly (P<0.05). Compared with the model group, the GLUT2and GCK mRNA expression of the electro-acupuncture on Yishu group and the electro-acupuncture on Pishu group rose significantly(P<0.05). The GLUT2mRNA expression of the electro-acupuncture on Yishu group is significantly higher than that of the electro-acupuncture on Pishu group (P<0.05).Conclusions:1. Electro-acupuncture on Yishu can adjust the blood glucose and lipid metabolism of T2DM rats, can improve their insulin sensitive index and their insulin resistance condition, can improve their damaged islet morphology, can boost their expression of PDX-1,GLUT2and GCK mRNA in the islet beta cells.2. Electro-acupuncture on Pishu can lower the level of blood glucose, can improve their insulin sensitive index and their insulin resistance condition,can boost their expression of PDX-1, GLUT2and GCK mRNA in the islet beta cells.3. The curative effects of the electro-acupuncture on Yishu and Pishu on T2DM rats may be related with the PDX-1-GLUT2-GCK pathway to boost the glucose metabolism in cells, and GLUT2plays an important role in this pathway.4. Both electro-acupuncture on Yishu and Pishu have certain effects on the type2diabetes, while electro-acupuncture on Yishu is slightly better in lowering the level of blood glucose,regulating lipid metabolism and GLUT2gene expression. That is to say, the curative effects of back shu points have certain relation with spinal segments, but Yishu has its specificity in treating diabetes, and the secific machanism still needs futhur exploration. |
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