吡非尼酮对肾纤维化大鼠的治疗作用及分子机制  

Therapeutic effect and molecular mechanism of pirfenidone on the renal fibrosis rats

作　　者：晏青 程芝梅 张帅 周石 YAN Qing;CHENG Zhimei;ZHANG Shuai;ZHOU Shi(School of Medical Imaging,Guizhou Medical University,Guiyang 550004,Guizhou,China)

机构地区：[1]贵州医科大学医学影像学院,贵州贵阳550004

出　　处：《贵州医科大学学报》2024年第3期354-360,共7页Journal of Guizhou Medical University

基　　金：国家自然科学基金(81760325)。

摘　　要：目的探讨吡非尼酮(PFD)对肾纤维化大鼠肾脏的治疗作用及机制。方法30只SD大鼠随机均分为对照组、模型组及治疗组,后2组大鼠腹腔注射50%四氯化碳(CCl_(4))油溶液建立肾纤维化模型,对照组腹腔注射等体积橄榄油,持续5周;造模结束,治疗组大鼠PFD水溶液灌胃给药,模型组和对照组大鼠同剂量生理盐水灌胃,持续4周;干预期间每天观察大鼠活动、进食饮水、毛发颜色以及大小便情况,于干预前以及干预第2、5、7及9周最后1次给药24 h后对大鼠进行称重并记录大鼠体质量及一般情况;干预第9周末处死各组大鼠,取心脏血检测血清尿素氮(BUN)、血肌酐(Scr)及尿酸(UA)含量,取肾脏组织采用苏木素伊红染色(HE)和Masson染色观察各组大鼠肾组织损伤和纤维化程度,采用蛋白免疫印迹法检测各组大鼠肾脏组织中沉默信息调节因子3(SIRT3)、缺氧诱导因子-1α(HIF-1α)、转化生长因子-β1(TGF-β1)、α平滑肌肌动蛋白(α-SMA)、Ⅰ型胶原(ColⅠ)、Ⅲ型胶原(ColⅢ)、金属蛋白酶组织抑制因子1(TIMP1)及基质金属蛋白酶2(MMP2)蛋白的表达。结果与对照组比较,模型组大鼠肾功能损伤和纤维化明显,血清BUN、Scr及UA含量降低(P<0.05),肾组织中HIF-1α、TGF-β1、α-SMA、ColⅠ、ColⅢ及TIMP1蛋白表达增高(P<0.05),MMP2和SIRT3蛋白表达降低(P<0.05);与模型组比较,治疗组大鼠肾功能损伤和纤维化程度减轻,血清肾功能BUN、Scr、UA含量增高(P<0.05),肾组织中HIF-1α、TGF-β1、α-SMA、ColⅠ、ColⅢ及TIMP1蛋白表达降低(P<0.05),MMP2和SIRT3蛋白表达增高(P<0.05)。结论PFD可减轻肾纤维化大鼠肾功能损害和纤维化程度,其机制可能与上调SIRT3蛋白表达有关。Objective To investigate the therapeutic effect and mechanism of pirfenidone(PFD)on kidney of rats with renal fibrosis.Methods Thirty SD rats were randomly divided into control group,model group,and treatment group with equal number.The latter two groups were intraperitoneally injected with 50% CCl_(4) oil solution to establish a renal fibrosis model.The control group was intraperitoneally injected with the same dose of olive oil for 5 weeks.At the end of modeling,the treatment group rats were given PFD aqueous solution by gavage,while the model group and control group were given the same dose of normal saline by gavage for 4 weeks.During the experiment,the activity,food and water intake,hair color,and bowel movements of rats were observed daily.The rats were weighed,their body mass and general condition were recorded before the experiment,and on the 2 nd,5 th,7 th after intervention,and 24 h after the last dose of the 9 th week.At the end of the 9 th week of experiment,all groups of rats were euthanized.Heart blood was collected and tested for blood urea nitrogen(BUN),serum creatinine(Scr),and uric acid(UA)levels.Renal tissue was stained with Hematoxylin-Eosin(HE)and Masson's staining to observe the degree of renal tissue damage and fibrosis.Western Blot assay was adopted to test the expression of silence information regulator 3(SIRT3),hypoxia-inducible factor-1α(HIF-1α),transforming growth factor-β1 transforming growth factor-β1,(TGF-β1),Alpha-smooth muscle actin(α-SMA),Collagen typeⅠ(ColⅠ),Collagen typeⅢ(ColⅢ),tissue inhibitor of metalloproteinases 1(TIMP1),and Matrix metalloproteinases 2(MMP2).Results Compared with the control group,the model group showed significant renal function damage and fibrosis;serum BUN,SCR,and UA decreased(P<0.05);the expression level of HIF-1α,TGF-β1,α-SMA,ColⅠ,ColⅢ,and TIMP1 protein in renal tissue increased(P<0.05),the expression level of MMP2 and SIRT3 protein decreased(P<0.05).Compared with the model group,the treatment group showed reduced renal function da

关 键 词：四氯化碳 缺氧诱导因子1 Α亚基 转化生长因子Β1 吡非尼酮 沉默信息调节因子3 肾纤维化 

分 类 号：R692[医药卫生—泌尿科学]