摘要
目的:观察温阳消癥方对5/6肾切除小鼠残肾功能及肾脏纤维化的影响,并探讨其作用机制。方法:随机从32只小鼠中取24只制作5/6肾切除慢性肾功能衰竭肾纤维化模型,2周后根据血清肌酐值将24只小鼠分为5/6肾切除组、代文组和温阳消癥方组,另8只作为假手术组。治疗8周后,Western blot检测各组小鼠肾组织JAK2、STAT3、p-JAK2、p-STAT3、转化生长因子β1(TGF-β1)、Smad3、I型胶原蛋白(Col-I)、Ⅲ型胶原蛋白(Col-Ⅲ)、α-SMA蛋白表达水平,免疫组织化学染色法测定小鼠肾组织Col-I、Col-Ⅲ、α-SMA水平,用RT-q PCR法测定TGF-β1、Smad3 mRNA水平。结果:治疗8周后,模型组小鼠肾组织JAK2、STAT3、pJAK2、p-STAT3、TGF-β1、Smad3、Col-I、Col-Ⅲ、α-SMA蛋白表达均较假手术组上调(P<0.01),模型组小鼠肾组织TGF-β1、Smad3 mRNA表达较假手术组上调(P<0.01);与模型组比较,代文组、温阳消癥方组JAK2、STAT3、p-JAK2、p-STAT3、TGF-β1、Smad3、Col-I、Col-Ⅲ、α-SMA蛋白表达和TGF-β1、Smad3 mRNA表达均下调(P<0.01);与代文组比较,除TGF-β1、α-SMA外,温阳消癥方组其余指标均下调(P<0.01)。结论:温阳消癥方能够保护5/6肾切除小鼠残肾功能,减轻肾纤维化病变程度,其机制可能与抑制TGF-β1/Smad3、JAK2/STAT3通路相关。
Objective: To observe the effect of Wenyangxiaozheng decoction on renal function and renal fibrosis in 5/6 nephrectomy(5/6N) mice and explore its mechanism. Methods: Out of 32 C57 mice 24 were randomly chosen to establish 5/6N model which closely simulated the process of human chronic renal failure and renal fibrosis. Two weeks after the operation, 24 mice were divided as 5/6N group, diovan group and wenyangxiaozheng decoction group according to the value of serum creatinine. The other 8 mice were treated as sham operation group. Eight weeks after the treatment, the protein expression of JAK2, STAT3, p-JAK2, pSTAT3, transforming growth factor-β1(TGF-β1), Smad3 was determined by Western blot;the protein expression of Collagen-I(Col-I), Collgaen-Ⅲ(Col-Ⅲ) and α-SMA was determined by both Western blot and immunohistochemistry, TGF-β1, Smad3 mRNA were determined by RT-qPCR. Results: Eight weeks after the treatment, the protein expression of JAK2, STAT3, p-JAK2, p-STAT3, TGF-β1, Smad3, Col-I, Col-Ⅲ, α-SMA and TGF-β1, Smad3 mRNA expression in 5/6N group was significantly upregulated when compared with that of sham group(P<0.01);compared with 5/6N group, the protein expression of JAK2, STAT3, p-JAK2, p-STAT3,TGF-β1, Col-I, Col-Ⅲ, α-SMA and TGF-β1, Smad3 mRNA expression was obviously downregulated in diovan group and wenyangxiaozheng decoction group(P<0.01);compared with diovan group, indexes were all downregulated in wenyangxiaozheng decoction group(P<0.01) except TGF-β1 and α-SMA. Conclusion:Wenyangxiaozheng decoction could protect renal function and reduce the degree of renal fibrosis in 5/6N mice,and the mechanism may be related to the inhibition of TGF-β1/Smad3 and JAK2/STAT3 signaling pathway.
作者
余柯娜
何立群
林晓蒙
蔡旭东
YU Kena;HE Liqun;LIN Xiaomeng;CAI Xudong(Department of Nephropathy,Ningbo Hospital of Traditional Chinese Medicine Affiliated to Zhejiang University of Traditional Chinese Medicine,Ningbo 315000,China;Department of Nephropathy,Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine,Shanghai 200021,China)
出处
《温州医科大学学报》
2022年第7期524-531,538,共9页
Journal of Wenzhou Medical University
基金
国家自然科学基金项目(81373615)
浙江省自然科学基金项目(11DZ1973100)
宁波市医疗卫生品牌学科建设项目(PPXK2018-07)
宁波市科技计划项目(2019A610363)
浙江省中医药科技计划项目(2021ZB251)。
