中华中医药杂志2026，Vol.41Issue(3)：925-930,6.
益气活血通络方调控NR4A1/Hippo/YAP途径抑制肾小管上皮细胞G2/M期阻滞拮抗肾纤维化的机制研究
Mechanism of Yiqi Huoxue Tongluo Formula on inhibiting G2/M arrest of renal tubule epithelial cells to ameliorate renal fibrosis by regulating NR4A1/Hippo/YAP
摘要
Abstract
Objective:To investigate the mechanism of Yiqi Huoxue Tongluo Formula regulating the solitary nucleus receptor 4A(NR4A1)/Hippo/Yes kinase-associated protein(YAP)pathway to inhibit the G2/M phase arrest of renal tubular epithelial cells(TEC)in unilateral ureteral obstruction(UUO)mice against renal fibrosis(RF).Methods:A total of 40 male C57/BL6 mice were randomly divided into Sham group(Sham group),model group(UUO group),Yiqi Huoxue Tongluo Formula group(YQHX group),NR4A1 agonist Csn-B group(Csn-B group),with 10 mice in each group.UUO modeling was performed in all groups except sham operation group.HE,Masson,and Sirius red staining were used to observe the pathological changes in the kidney tissues.Immunohistochemistry was employed to detect the expression of α-SMA and Vimentin.Western Blot was used to detect the expression of α-SMA,Vimentin,NR4A1,and proteins related to the Hippo/YAP pathway.Immunofluorescence staining was utilized to detect the co-localization of YAP and TEAD1,as well as Ki67 and p-H3.In vitro,HK-2 cells were randomly divided into blank group(Con group),TGF-β1 group(TGF group),TGF-β1+YQHX group(TGF+YQHX group)and TGF-β1+Csn-B group.Except for the blank group,cells in the other groups were stimulated with TGF-β1 to establish a cell fibrosis model,which was treated for 48 hours,cells were collected.Western Blot was used to detect the expression of α-SMA,Vimentin,NR4A1 and proteins related to the Hippo/YAP pathway.Immunofluorescence staining was used to detect the co-localization of YAP and TEAD1,as well as Ki67 and p-H3.Results:In vivo experiments:compared with the UUO group,the degree of renal fibrosis was alleviated in the YQHX group and the Csn-B group,the expression of α-SMA and Vimentin was significantly decreased(P＜0.05),while the expression of NR4A1 and the protein ratios of p-YAP/YAP,p-MST/MST and p-LATS/LATS was significantly increased(P＜0.05),the co-localization of YAP and TEAD1,as well as p-H3 and Ki67,was weakened.In vitro experiments:compared with the TGF group,the expression of α-SMA and Vimentin was significantly decreased in the TGF+YQHX group and the TGF+Csn-B group(P＜0.05),and the expression of NR4A1 and the protein ratios of p-YAP/YAP,p-MST/MST,and p-LATS/LATS were significantly increased(P＜0.05),the co-localization of YAP and TEAD1,as well as p-H3 and Ki67,was weakened.Conclusion:Yiqi Huoxue Tongluo Formula can antagonize RF by regulating NR4A1/Hippo/YAP pathway to inhibit the G2/M phase arrest of TEC.关键词
益气活血通络方/肾纤维化/单侧输尿管梗阻/NR4A1/Hippo/YAP途径/G2/M期阻滞Key words
Yiqi Huoxue Tongluo Formula/Renal fibrosis(RF)/Unilateral ureteral obstruction/NR4A1/Hippo/YAP pathway/G2/M phase block引用本文复制引用
赵晨晨,高兰君,方芳,许畅,翟楠,钟艳,王香婷..益气活血通络方调控NR4A1/Hippo/YAP途径抑制肾小管上皮细胞G2/M期阻滞拮抗肾纤维化的机制研究[J].中华中医药杂志,2026,41(3):925-930,6.基金项目
河北省中医药基金自然科学重点项目(No.H2022423351),河北省中医药联合基金重点项目(No.H2023423050),河北中医药大学研究生创新基金项目(No.XCXZZSS2024013) （No.H2022423351）
