痛泻要方合四逆散对糖尿病的治疗效果及作用机制研究

Experimental study on therapeutic effect and mechanism of Tongxie Yaofang combined with Sini powder on diabetes mellitus

目的探讨痛泻要方合四逆散对糖尿病小鼠的治疗作用及机制。方法将24只KK-Ay小鼠按照随机数字表分为模型组及低、中、高剂量中药组4组,各6只;另设6只C57BL/6J小鼠作为空白组。低、中、高剂量组分别给予中药生药量1.84、3.68、5.52 g/(kg·d),空白组、模型组予等体积蒸馏水,1次/d,连续给药4周。比较各组体质量、空腹血糖(FBG)、胰岛素(INS)、胰岛素抵抗指数(HOMA-IR)、皮质酮(CORT)变化;Western blotting法检测各组海马组织11β-羟基类固醇脱氢酶Ⅰ(HSD1)、糖皮质激素受体(GR)蛋白表达水平变化。结果与空白组比较,干预第0、1、2、3、4周模型组体质量高(P<0.05)。与模型组比较,干预第1、2周,低、中、高剂量组体质量低,差异无统计学意义(P>0.05);第3、4周时,低、中、高剂量组体质量低,差异有统计学意义(P<0.05)。与空白组比较,干预第0、1、2、3、4周模型组FBG高(P<0.05)。与模型组比较,干预第0、1、2、3周,低、中、高剂量组FBG水平低,差异无统计学意义(P>0.05);第4周,高剂量组FBG水平低,差异有统计学意义(P<0.05)。与空白组比较,模型组INS水平及HOMA-IR高(P<0.05)。与模型组比较,低剂量组INS水平及HOMA-IR低,差异无统计学意义(P>0.05);中、高剂量组INS水平及HOMA-IR低,差异有统计学意义(P<0.05)。与模型组比较,低剂量组CORT水平低,差异无统计学意义(P>0.05);中、高剂量组CORT低,差异有统计学意义(P<0.05)。与空白组比较,模型组海马组织11β-HSD1蛋白表达高、GR蛋白表达低,差异有统计学意义(P<0.05)。与模型组比较,低剂量组海马组织11β-HSD1蛋白表达低、GR蛋白表达高,差异均无统计学意义(P>0.05);中、高剂量组海马组织11β-HSD1蛋白表达低、GR蛋白表达高,差异均有统计学意义(P<0.05)。结论痛泻要方合四逆散能改善2型糖尿病小鼠FBG水平和胰岛素抵抗状态,其作用机制可能与降低皮质酮含量及调控海马组织11β-HSD1及GR蛋白表达有关。

Objective To observe the therapeutic effect and mechanism of Tongxie Yaofang combined with Sini powder on diabetes mellitus.Methods 24 KK-Ay mice were randomly divided into model group and low,medium and high dose Chinese medicine treatment groups respectively of 6 mice in each group,and another 6 C57BL/6J mice were taken as blank control group.The low,middle and high dose groups were given the crude dosage of Chinese medicine 1.84,3.68 and 5.52 g/(kg d)respectively,while the blank group and model group were given the same volume of distilled water once a day for 4 weeks.The changes of body mass,fasting blood glucose(FBG),insulin(INS),insulin resistance index(HOMA-IR)and corticosterone(CORT)in each group were compared.The expression levels of 11βhydroxysteroid dehydrogenaseⅰ(HSD1)and glucocorticoid receptor(GR)in hippocampus were detected by Western blotting.Results Compared with the blank group,the body mass of the model group was higher at the 0,1,2,3 and 4 weeks of intervention(P<0.05).Compared with the model group,the body weight of the low,medium and high dose groups was lower in the first and second weeks of intervention(P>0.05).At the 3rd and 4th week,the body weight of the low,middle and high dose groups was low(P<0.05).Compared with the blank group,the FBG in the model group was higher at the 0,1,2,3 and 4 weeks of intervention(P<0.05).Compared with the model group,the level of FBG in the low,middle and high dose groups was lower at the 0,1,2 and 3 weeks of intervention(P>0.05).At the 4th week,the level of FBG in the high dose group was low(P<0.05).Compared with the blank group,the INS level and HOMA-IR in the model group were higher(P<0.05).Compared with the model group,the INS level and HOMA-IR in the low-dose group were lower,with no statistical significance(P>0.05).The levels of INS and HOMA-IR in middle and high dose groups were lower,and the difference was statistically significant(P<0.05).Compared with the model group,the CORT level in the low-dose group was lower,and the difference was not statistically significant(P>0.05).CORT in middle and high dose groups was lower,and the difference was statistically significant(P<0.05).Compared with the blank group,the expression of 11β-HSD1 protein in hippocampus of model group was higher,while the expression of GR protein was lower,with statistical significance(P<0.05).Compared with the model group,the expression of 11β-HSD1 protein in hippocampus of the low-dose group was lower,while the expression of GR protein was higher,with no statistical significance(P>0.05).The expression of 11β-HSD1 protein in hippocampus was low and GR protein was high in middle and high dose groups,and the difference was statistically significant(P<0.05).Conclusion Tongxie Yaofang combined with Sini Powder can decrease the level of FBG and improve insulin resistance in type 2 diabetes mice,and its mechanism may be related to the reduction of corticosterone content and the regulation of 11β-HSD1and GR protein expression.