参芪补胰方通过调节TLR4/MyD88/NF-κB信号通路治疗2型糖尿病大鼠的作用机制

Mechanism of Shenqi Buyi Decoction(参芪补胰方)Regulating TLR4/MyD88/NF-B Signaling Pathway to Treat Rats of Type 2 Diabetes

目的观察参芪补胰方对2型糖尿病(type 2 diabetes mellitus,T2DM)大鼠TLR4/MyD88/NF-κB信号通路的影响,探讨参芪补胰方治疗T2DM的作用机制。方法采用随机对照法将48只SD大鼠随机分为7组,正常组予以基础饲料喂养,其余各组动物采用高糖高脂饲养加腹腔注射STZ法造模,二甲双胍组给予盐酸二甲双胍片,中药组给予不同剂量参芪补胰方,给药6周后留取标本。ELISA法检测大鼠肿瘤坏死因子-α(tumor necrosis factor,TNF-α)、白细胞介素-1β(Interleukin 1β,IL-1β)、白细胞介素-10(Interleukin 10,IL-10)、白细胞介素-17(Interleukin 17,IL-17)水平,HE染色法观察大鼠胰腺组织病理变化,TUNEL染色法观察大鼠胰岛细胞凋亡情况,RT-PCR法检测大鼠胰腺组织TLR4、MyD88、NF-κB的mRNA表达水平,Western blot法检测大鼠胰腺组织TLR4、MyD88、NF-κB的蛋白表达水平。结果参芪补胰方对T2DM FBG水平方面有明显下降作用,与模型组比较有统计学差异(P<0.05);参芪补胰方组大鼠血清TNF-α、IL-1β、IL-10、IL-17表达水平明显降低(P<0.05);可以通过改善大鼠胰腺组织损伤、促进细胞结构组织修复、减少细胞凋亡起到抗T2DM的作用;同时,与模型组比较,参芪补胰方组大鼠胰腺组织TLR4、MyD88、NF-κB mRNA及蛋白表达均明显降低(P<0.05)。结论参芪补胰方可能通过调控T2DM大鼠炎症因子及TLR4、MyD88、NF-κB表达发挥其抗T2DM的作用。

Objective The effect of Shenqi Buyi Decoction(参芪补胰方)on TLR4/MyD88/NF-B signaling in type 2 diabe⁃tes mellitus(T2DM)rats was observed,and the mechanism of Shenqi Buyi Decoction in the treatment of T2DM was investigated.Methods Forty-eight SD rats were randomly divided into 7 groups.The normal group were fed with basic feed,the rest of the an⁃imals were fed with high sugar and high fat and injected with STZ intraperitoneally to make models.Metformin Hydrochloride Tablets were given to the metformin group,and different doses of Shenqi Buyi Decoction were given to the traditional Chinese medicine group,and samples were taken out 6 weeks after administration.ELISA method was used to detect the levels of tumor necrosis factor(TNF-α),interleukin 1β(IL-1β),interleukin 10(IL-10)and interleukin 17(IL-17).HE staining method was used to observe the pathological changes of pancreas in rats.TUNELstaining method was used to observe the islet cell apop⁃tosis.RT-PCR method was used to detect the expressions of TLR4,MyD88 and NF-κB mRNA in pancreatic tissue of the rats.Western blot was used to detect the expressions of TLR4,MyD88 and NF-κB proteins in pancreatic tissue of the rats.Re⁃sults Shenqi Buyi Decoction decreased fast blood glucose(FBG)levels in DM rats compared with the model group(P<0.05).The levels of TNF-α,IL-1β,IL-10 and IL-17 were significantly decreased in the Shenqi Buyi Decoction group(P<0.05).It can play an anti-T2DM role by improving rat pancreatic tissue damage,promoting cell structural tissue repair and reducing cell apoptosis.Meanwhile,when compared with those of the model group,the expressions of TLR4,MyD88 and NF-κB mRNA and proteins were significantly decreased in the pancreatic tissues in the Shenqi Buyi Decoction group(P<0.05).Conclusion It is possible that Shenqi Buyi Decoction may exert its anti-T2DM effect by regulating inflammatory factors and expressions of TLR4,MyD88 and NF-κB in T2DM rats.