化瘀通络中药通过抑制TGF-β1/ERK通路减轻糖尿病肾病大鼠肾纤维化研究  

Huayu Tongluo Chinese medicines alleviate renal fibrosis by inhibiting TGF-β1/ERK pathway in diabetic nephropathy rats

作　　者：郭帅 方敬[1] 杨帆[1] 李雅纯 陈志强 GUO Shuai;FANG Jing;YANG Fan;LI Ya-chun;CHEN Zhi-qiang(Hebei University of Chinese Medicine,Shijiazhuang 050000,China;Hebei Provincial Hospital of TCM,Shijiazhuang 050000,China)

机构地区：[1]河北中医学院,河北石家庄050000 [2]河北省中医院,河北石家庄050000

出　　处：《中草药》2021年第10期2996-3001,共6页Chinese Traditional and Herbal Drugs

基　　金：国家自然科学基金资助项目(81373804)。

摘　　要：目的考察化瘀通络中药对糖尿病肾病大鼠肾纤维化的治疗作用,并基于转化生长因子-β1(transforminggrowth factor-β1,TGF-β1)/细胞外信号调节蛋白激酶(extracellular regulated protein kinase,ERK)信号通路探讨其作用机制。方法随机选取10只SD大鼠作为对照组,其余大鼠ip链脲佐菌素(40mg/kg)联合高脂饲料喂养建立糖尿病模型,糖尿病模型大鼠随机分为模型组和化瘀通络中药组。给予化瘀通络中药第0、4、8、12、16周末,大鼠尾静脉采血检测空腹血糖;给予化瘀通络中药16周后,检测各组大鼠血清中肌酐、尿素氮和尿酸水平;考察各组大鼠肾组织病理变化;采用免疫组化法检测肾组织III型胶原(collagen III,Col-III)和TGF-β1表达;采用qRT-PCR法检测肾组织TGF-β1 mRNA表达;采用Western blotting法检测肾组织c-Raf、磷酸化c-Raf(p-c-Raf)、丝裂原活化细胞外信号调节蛋白激酶(mitogen-activatedextracellular signal regulated kinase,MEK)、p-MEK、ERK和p-ERK蛋白表达。结果与模型组比较,化瘀通络中药组大鼠血清中肌酐和尿素氮水平显著降低(P<0.01),肾组织病理损伤和纤维化减轻,肾组织Col-III和TGF-β1沉积减少,TGF-β1 mRNA表达水平明显降低(P<0.05),p-c-Raf、p-MEK和p-ERK蛋白表达水平均显著降低(P<0.05)。结论化瘀通络中药能够减轻糖尿病肾病大鼠肾组织病理损伤,改善肾纤维化,其机制可能与抑制TGF-β1/ERK信号通路相关。Objective To investigate the therapeutic effect of Huayu Tongluo Chinese medicines(removing blood stasis and dredging collaterals)on renal fibrosis in diabetic nephropathy rats,and explore its mechanism based on transforming growth factor-β1(TGF-β1)/extracellular signal regulated protein kinase(ERK)signaling pathway.Methods Ten SD rats were randomly selected as control group,the rest rats were ip administrated streptozotocin(40 mg/kg)and fed with high-fat diet to establish a diabetes model.Diabetes model rats were randomly divided into model group and Huayu Tongluo Chinese medicines group.Rats were given Huayu Tongluo Chinese medicines on 0 th,4 th,8 th,12 th,and 16 th weekends,and blood was collected from tail vein to detect fasting blood glucose;On 16 th week after given Huayu Tongluo herbs,levels of creatinine,urea nitrogen and uric acid in serum of rats were measured.Pathological changes in kidney tissue of rats in each group was investigated.Immunohistochemistry was used to detect the expression of collagen III(Col-III)and TGF-β1 in kidney tissue.TGF-β1 m RNA level in renal tissue was detected by q RT-PCR.Expressions of c-Raf,phosphorylated c-Raf(p-c-Raf),mitogen-activated extracellular signal regulated kinase(MEK),p-MEK,ERK and p-ERK were detected by Western blotting.Results Compared with model group,levels of creatinine and urea nitrogen in serum of rats in Huayu Tongluo Chinese medicines group were significantly reduced(P<0.01),renal tissue pathological damage and fibrosis were reduced,deposition of Col-III and TGF-β1 in renal tissue was reduced,TGF-β1 m RNA level was significantly reduced(P<0.05),protein expressions of p-c-Raf,p-MEK and p-ERK were significantly reduced(P<0.05).Conclusion Huayu Tongluo Chinese medicines can reduce pathological damage of kidney in diabetic nephropathy rats and improve renal fibrosis,and its mechanism may be related to inhibition of TGF-β1/ERK signaling pathway.

关 键 词：化瘀通络中药 糖尿病肾病 肾损伤 肾纤维化 TGF-β1/ERK信号通路 

分 类 号：R285.5[医药卫生—中药学]