益气活血通络方抑制TNF-α/Caspase-3/GSDME介导的巨噬细胞焦亡改善肾纤维化  

Yiqi Huoxue Tongluo Recipe inhibiting TNF-α/Caspase-3/GSDME mediated macrophage apoptosis to ameliorate renal fibrosis

作　　者：王洪双 方芳 赵晨晨 高兰君 王筝 钟艳 王香婷 WANG Hongshuang;FANG Fang;ZHAO Chenchen;GAO Lanjun;WANG Zheng;ZHONG Yan;WANG Xiangting(Hebei University of Chinese Medicine,Shijiazhuang 050200,China;Hebei Key Laboratory of Integrative Medicine on Liver-Kidney Patterns Research,Shijiazhuang 050091,China;Institute of Integrative Medicine,Hebei University of Chinese Medicine,Shijiazhuang 050091,China)

机构地区：[1]河北中医药大学,石家庄050200 [2]河北省中西医结合肝肾病证研究重点实验室,石家庄050091 [3]河北中医药大学中西医结合研究所,石家庄050091

出　　处：《中华中医药杂志》2024年第8期4327-4331,共5页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：河北省中医药联合基金重点项目(No.H2022423351);河北中医药大学2023年研究生创新资助项目(No.XXXZZBS2023002);河北省中医药联合基金培育项目(No.H2023423050)。

摘　　要：目的:观察益气活血通络方通过抑制半胱氨酸天冬氨酸蛋白酶-3/膜穿孔蛋白E(Caspase-3/GSDME)介导的巨噬细胞焦亡对肾纤维化的影响。方法:将36只SPF级雄性C57BL/6J小鼠随机分为假手术组、单侧输尿管梗阻(UUO)组、益赛普组、益气活血通络方组,每组9只。除假手术组外,其余各组小鼠予以UUO造模。加治疗过程14 d后取材行HE、Masson、天狼星红染色观察肾脏组织病理形态;免疫荧光双重染色检测F4/80与GSDME的共表达,Western Blot检测α-SMA、Vimentin、CollagenⅠ、Caspase-3、GSDME、肿瘤坏死因子-α(TNF-α)的蛋白表达。结果:与假手术组比较,UUO组肾小管扩张、萎缩、炎症细胞浸润及胶原沉积增多;α-SMA、Vimentin、CollagenⅠ、Cleaved Caspase-3、GSDME N-端(GSDME-N)、TNF-α表达显著升高,F4/80与GSDME双重染色共表达显著增多(P<0.05)。与UUO组比较,各治疗组肾小管损伤程度明显改善,胶原沉积减少,F4/80与GSDME的双重染色表达明显减少,α-SMA、Vimentin、CollagenⅠ、Cleaved Caspase-3、GSDME-N、TNF-α表达显著降低(P<0.05)。结论:益气活血通络方可抑制TNF-α/Caspase-3/GSDME介导的巨噬细胞焦亡改善肾纤维化。Objective:To observe the effect of Yiqi Huoxue Tongluo Recipe on renal fibrosis by regulating the apoptosis of macrophages mediated by Caspase-3/GSDME.Methods:Thirty-six SPF male C57BL/6J mice were randomly divided into sham surgery group,unilateral ureteral obstruction(UUO)group,Etanercept group,Yiqi Huoxue Tongluo Recipe group,with 9 mice in each group.Except for the sham surgery group,all other groups of mice were subjected to UUO modeling.After 14 days of treatment,samples were taken for HE,Masson,and Sirius red staining to observe the pathological morphology of kidney tissue.Immunofluorescence double staining for co-expression of F4/80 and GSDME,Western Blot detectedα-SMA,Vimentin,CollagenⅠ,Caspase-3,GSDME,TNF-αprotein expression.Results:Compared with the sham surgery group,the UUO group showed increased renal tubular dilation,atrophy,inflammatory cell infiltration,and collagen deposition;the expression ofα-SMA,Vimentin,CollagenⅠ,Cleaved Caspase-3,GSDME-N,TNF-αwas significantly increased,and the co-expression of F4/80 and GSDME double staining was significantly increased(P<0.05).Compared with the UUO group,the degree of renal tubular injury in each treatment group was significantly improved,collagen deposition was reduced,and the double staining expression of F4/80 and GSDME was significantly reduced,the expression ofα-SMA,Vimentin,CollagenⅠ,Cleaved Caspase-3,GSDME-N,TNF-αwere significantly reduced(P<0.05).Conclusion:Yiqi Huoxue Tongluo Recipe can inhibit TNF-α/Caspase-3/GSDME mediated macrophage apoptosis and improve renal fibrosis.

关 键 词：益气活血通络方 单侧输尿管梗阻 肿瘤坏死因子-α 巨噬细胞焦亡 半胱氨酸天冬氨酸蛋白酶-3 膜穿孔蛋白E 

分 类 号：R285.5[医药卫生—中药学]