白首乌C21甾苷通过TLR4通路防治大鼠肝肾纤维化的作用研究  

Effect of C21 steroidal glycoside of Cynanchum auriculatum on liver and kidney fibrosis through TLR4 pathway

作　　者：庄子锐 王明亮 彭蕴茹[1,2] 沈明勤[1,2] ZHUANG Zi-rui;WANG Ming-liang;PENG Yun-ru;SHEN Ming-qin(Affiliated Hospital of Integrated Traditional Chinese and Western Medicine,Nanjing University of Chinese Medicine,Nanjing 210028,China;Jiangsu Province Academy of Traditional Chinese Medicine,Nanjing 210028,China)

机构地区：[1]南京中医药大学附属中西医结合医院,江苏南京210028 [2]江苏省中医药研究院,江苏南京210028

出　　处：《中国中药杂志》2021年第11期2857-2864,共8页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金项目(81774178,81373888);江苏省研究生实践创新计划项目(SJCX20_0572)。

摘　　要：采用硫代乙酰胺(thioacetamide, TAA)和结扎单侧输尿管法(unilateral ureteral ligation, UUO)建立SD大鼠肝肾纤维化复合模型,模型建立的同时开始分组给药。造模大鼠随机分为3组,即模型组和白首乌C21甾苷低、高剂量组,另设空白组,连续给药4周。4周后取血清检测肝肾功能的生化指标,取尿液进行尿蛋白、尿肌酐检测,取肝、肾组织样本进行HE和Masson染色及羟脯氨酸含量测定,利用Western blot法检测纤维化相关指标、炎症因子和TLR4通路相关蛋白的表达情况,观察白首乌C21甾苷对肝肾纤维化的防治作用,并探索其分子机制。4周后模型组大鼠血清生化结果显示肝、肾功能受到严重损伤,并且病理切片显示肝、肾组织炎症细胞浸润、实质细胞减少、间质纤维化增多,肝肾纤维化复合模型建立成功;而白首乌C21甾苷低、高剂量(150,300 mg·kg^(-1))组表现出对肝肾纤维化进程明显的干预作用,与模型组相比白首乌C21甾苷可明显降低组织内胶原沉积、减轻肝肾纤维化的病变程度;同时实验发现模型组大鼠肝、肾组织中TLR4和MYD88信号通路蛋白表达显著上升,NF-κB信号通路蛋白大量向核内迁移,而白首乌C21甾苷低、高剂量组TLR4和MYD88信号通路蛋白表达以及NF-κB向核内迁移被显著抑制。因此推测白首乌C21甾苷可通过抑制TLR4/MYD88/NF-κB信号通路防治肝肾纤维化。The liver and kidney fibrosis model was established by thioacetamide(TAA) and unilateral ureteral obstruction(UUO) in SD rats. The rats were randomly divided into three groups: model group, low and high-dose groups of C21 steroidal glycosides of Cynanchum auriculatum. Another blank control group was set. Four weeks later, serum was taken to detect the biochemical indexes of liver and kidney function. Urine protein and urine creatinine were detected by kits. Liver and kidney tissue samples were stained with HE and Masson staining, and hydroxyproline content was detected. Western blot was used to detect expressions of fibrotic proteins, inflammatory factors and TLR4 signaling pathways, so as to observe the preventive and therapeutic effects of C21 steroidal glycosides from C. auriculatum on hepatic and renal fibrosis and explore its molecular mechanism. Four weeks later, serum biochemical results showed that liver and kidney functions were seriously damaged, and pathological sections showed that inflammatory cell infiltration, decrease of parenchymal cells, and increase of interstitial fibrosis in liver and kidney tissues. The results showed that low and high doses(150, 300 mg·kg^(-1)) of C21 steroidal glycosides could significantly reduce the collagen deposition and the pathological changes of liver and kidney fibrosis compared with the model group. At the same time, we found that the expression levels of TLR4 and MyD88 signaling pathway proteins were significantly increased in the liver and kidney tissues of the model group, and a large number of NF-κB signaling pathway proteins migrated into the nucleus. On the contrary, the expression levels of TLR4, MyD88 signaling pathway proteins and the nuclear migration of NF-κB were significantly inhibited in the low and high dose groups of C21 steroidal glycosides from C. auriculatum. Therefore, it was speculated that the mechanism of C21 steroidal glycoside for preventive and therapeutic effect on hepatic and renal fibrosis was related to inhibit TLR4/MYD88/NF-κB in

关 键 词：白首乌C21甾苷 肝肾纤维化 胶原沉积 炎症因子 TLR4通路 

分 类 号：R285.5[医药卫生—中药学]