基于JNK传导通路探析解毒通络调肝方调控内质网应激治疗2型糖尿病的实验研究

Experimental study on Jiedu Tongluo Tiaogan Formula to regulate and control endoplasmic reticulum stress in the treatment of type 2 diabetes: an exploration based on JNK signaling pathway

观察解毒通络调肝方对肝脏JNK传导途径的影响。方法选用50只ZDF大鼠,饲养12周后48只成模,随机分组为模型组12只、二甲双胍组9只、解毒通络调肝方高、中、低剂量组各9只,ZL大鼠10只为空白对照组。模型组与空白组以蒸馏水,给药组给予相应药物,灌胃12周。定期对大鼠进行糖耐量实验,24周末切除肝脏,用于Western blot检测。结果与模型组比较,解毒通络调肝方高、中、低计量组和二甲双胍组各时点OGTT水平差异均有统计学意义(P<0.01);解毒通络调肝方高剂量与低剂量组比较,各时点差异均有统计学意义(P<0.05)。解毒通络调肝方高、中、低剂量组和二甲双胍组稳态葡萄糖输注率均较模型组提高,差异有统计学意义(P<0.01)。解毒通络调肝方高、中、低剂量组和二甲双胍组JNK及p-JNK表达量均较模型组下降,差异有统计学意义(P<0.01)。结论解毒通络调肝方改善治疗糖尿病可能的机制之一是抑制了JNK信号通路,降低了JNK磷酸化水平;糖毒性和胰岛素抵抗可能通过炎症激活JNK信号通路,增强JNK活性导致肝细胞损害。

Objective To observe the effect of Jiedu Tongluo Tiaogan Formula on liver JNK signaling pathway. Methods 50 ZDF rats were selected and fed for 12 weeks. Of them,48 rats were established into the models. They were randomly divided into the model group of 12 rats,metformin group of 9 rats,high-dose,moderate-dose and low-dose Jiedu Tongluo Tiaogan group of 9 rats each,blank control of 10 ZL rats. The model group and the blank group were given distilled water, drug taking group were given corresponding drugs by gavage for 12 weeks. Glucose tolerance test were made regularly. At the end of 24 th week,the rats＇ livers were removed for Western blot test. Results Compared with the model group,the difference of OGTT level at different time points in highdose,moderate-dose and low-dose Jiedu Tongluo Tiaogan Formula group and metformin group were statistically significant（ P 0. 01）;to compare high-dose and low-dose Jiedu Tongluo Tiaogan Formula, the difference of OGTT level at different time points were statistically significant（ P 0. 05）; steady state glucose infusion rate in high-dose,moderate-dose and low-dose Jiedu Tongluo Group and metformin group were increased compared with the model group,differences were statistically significant（ P 0. 01）. JNK and p-JNK expression in high-dose,moderate-dose and low-dose group and metformin group were lowered compared with the model group,the differences were statistically significant（ P 0. 01）. Conclusion One of the mechanisms of Jiedu Tongluo Tiaogan Formula to improve diabetes is to inhibit JNK signaling pathway,lower JNK phosphorylation level; glucose toxicity and insulin resistance may activate JNK signaling pathway and strengthen JNK activity which result in liver cell damage.