| Objective:This study was designed to observe effects on T2DM rats’ weight, fasting blood glucose, plasma lipids, serum insulin, diabetes-related indices, morphology of pancreatic islet cells, and gene expression levels in PXD-1, GLUT2, GCK and Insulin which are those of (3cells by EA on "Yi Shu" which are related to pancreas by nerve segment and on "Shen Shu" which are not related to pancreas by nerve segment as a control acupuncture point. And it was to verify its effects of treatment and possible mechanism of T2DM, and special effects and possible mechanisms on visceral functions by EA on acupuncture point related to visceral by nerve segment on.Methods:(1)By feeding high fat and sugar diet including intraperitoneal injection of STZ35mg/kg (2%concentration), T2DM rat model was established.704-week-old male Wistar rats were randomly divided into control group (n=8) and T2DM model group (n=62). Control group was fed normal diet; T2DM model group was fed high fat and sugar.4weeks later, T2DM model group was implemented a single intraperitoneal injection of STZ (35mg/kg), and After72hours of this injection FBG was tested at tail vein. In the condition that FBG≥11.1mmol/L, the rat oral glucose tolerance test (OGTT) was experimented after1week of injection of STZ. If2hPG at OGTT≥11.1mmol/L and there is high frequency of occurrence of polydipsia, urinary symptoms, then T2DM model might be successfully established.(2)51rats were successfully passed this test, and were randomly divided as these5types:One type is a model group of10rats, another type is a group with EA "Yi Shu" of11rats, the third type is a group with EA "Shen shu" of11rats, the last type is a group with a glimepiride by gavages of10rats. There is no intervention in a control group and a model group, and however, there are interventions in the others group giving EA or feeding gavages.(3) After four-weeks-intervention, specimens of rats were collected. Colorimetry of their serum was taken to test HDL-C, LDL-C, TG, TC, GSP, and other biochemical indicators. Radioimmunoassay of serum to test insulin concentrations and then insulin sensitivity index (ISI), HOMA insulin resistance index (HOMA-IR), and pancreatic β-cell function index (HOMA-β)was calculated by the result of serum insulin and FBG. Real-time PCR of it to verify the mRNA expression levels of PXD-1, GLUT2, GCK and Insulin. Results:(1) Efficacy of indicators on results show:①FBG:After the last intervention, compared with the model group FBG in "Yi shu" Group was decreased significantly (P<0.01), in "Shen shu" group and "Glimepiride" group it was also decreased, but not statistically significant (P>0.05).②GSP:Between the model group and three intervention groups, GSP was no significant difference (P>0.05).③Lipids:Compared with the model group,"Yi shu" group of TC, LDL-C, TG levels were decreased significantly (P respectively, P>0.05, P<0.05, P<0.01)."Shen shu" group of those levels were elevated, but not statistically significant (P>0.05)."Glimepiride" group of those levels were decreased, but not statistically significant (P>0.05).④HOMA-IR:Compared with the model group, Yi shu" group of HOMA-IR was significantly reduced (P<0.01)."Glimepiride" group of it was significantly lower (P<0.05).⑤HOMA-β: Compared with the model group,"Yi shu" group of HOMA-β was significantly elevated (P<0.01), no statistically significant difference with "Shen shu" group and "Glimepiride" group (P>0.05).⑥ISI:Compared with the model group, ISI index of "Yi shu" group and "Glimepiride" group was significantly increased (P<0.01, P<0.05).⑦HE:Islet structure of control group was full and large, oval or cylindrical-shaped, uniformly distributed within the islet cell nucleus, tightly packed, and abundant of cytoplasm. That of the model group, however, was irregular, smaller size, sparse arrangement of nuclei within islets, and there were decreased cell, cell swelling, necrosis and degeneration in the cytoplasm. Compared with the model group, in three intervention groups the volume increased, the number of islet cell nucleus increased, and cytoplasmic vacuoles degeneration reduced(2) The mechanism of indicators on results show:①PDX-1mRNA:Compared with the model group,"Yi shu" group and "Shen shu" group, and "Glimepiride" group of PDX-1mRNA expression were significantly increased (P<0.01), three intervention group have no significant difference between the groups compared (P>0.05).②GLUT2mRNA:Expression levels of the three intervention groups were significantly elevated compared with the model group (P<0.01)."Yi shu" group compared with other two groups, of GLUT2expression level was significantly increased (P<0.05).③GCK mRNA:Compared with the model group, Expression levels of the three intervention groups was significantly increased (P<0.01). No significant difference between the three intervention groups (P>0.05).④Insulin mRNA:Compared with the model group, the expression of three intervention group were significantly increased (P<0.01)."Yi shu" group and "Glimepiride" group of expression level was significantly higher than "Shen shu" group (P<0.01), while "Yi shu" group and "Glimepiride" group of expression level has no significant difference (P>0.05).Conclusion:(1) EA on "Yi Shu" which are related to pancreas by nerve segment had significant hypoglycemic effects. Only if the effect of acupuncture is cumulated by certain period, it would play a role to a certain extent. And EA on "Shen Shu" which are not related to pancreas by nerve segment as a control acupuncture point or other oral glimepiride did not have obvious hypoglycemic effects.(2) EA on "Yi Shu" had obvious lipid-lowering effects, but EA on "Shen shu" or other oral glimepiride did not.(3) EA on "Yi shu" in T2DM of rat had obvious improvements on IR, decreased insulin sensitivity, and pancreatic0-cell dysfunction. Oral glimepiride had obvious improvements on IR, decreased insulin sensitivity. EA on "Shen shu" did not have obvious improvements on IR, decreased insulin sensitivity, and pancreatic β-cell dysfunction.(4)EA on "Yi Shu","Shen shu" and other oral glimepiride had significant improvement on pancreatic β cells morphology.(5) EA on "Yi Shu","Shen shu" and other oral glimepiride had obvious improvement on the increase of pancreatic β cells PDX-1, GLUT2, GCK expression level, thereby increasing the expression level of Insulin gene and the secretion level of Insulin, while there were significant improvements on cell morphology. Among those three groups, EA on "Yi Shu" which are related to pancreas by nerve segment had significantly specific effects on the increases of GLUT2mRNA expression level. |
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