Zhou Xiangjun, Bai Chen, Sun Xinbo, Gong Xiaoxin, Yang Yong, Chen Congbo, Shan Guang, Yao Qisheng
a Department of Urology , Taihe Hospital, Hubei University of Medicine , Hubei , China.
b Department of General Surgery , Taihe Hospital, Hubei University of Medicine , Hubei , China.
Ren Fail. 2017 Nov;39(1):423-431. doi: 10.1080/0886022X.2017.1305409.
Puerarin (PR) is an isoflavonoid isolated from the root of the plant Pueraria lobata and has been widely used in traditional Chinese herbal medicine for the treatment of various diseases. Oxidative stress and epithelial cell apoptosis play important roles in the renal fibrotic process. The present study aimed to determine whether or not PR inhibits renal fibrosis by reducing oxidative stress induced-epithelial cell apoptosis. In vivo, unilateral ureteral obstruction (UUO) induced renal fibrosis, and epithelial cell apoptosis. A total of 24 mice were randomly assigned to four experimental groups: sham, UUO alone, UUO +50 mg/kg PR, and UUO +100 mg/kg PR. In vitro, 200 μM hydrogen peroxide (HO) induced epithelial cell apoptosis. The experiments were dived into four groups: control, HO alone, HO+50 μM PR, and HO+100 μM PR. Tubular injury was measured in the renal cortex of the mice through periodic acid-Schiff (PAS) staining, and the extracellular matrix (ECM) was measured through Sirius red (SR), immunohistochemistry (IHC) staining, and Western blot. Renal epithelial cell apoptosis was measured through terminal deoxynucleotidyl transferase-mediated dUTP Nick-End labeling (TUNEL), flow cytometry (FCM), and Hoechst assays. The protein expression of NOX4, caspase3, ERK, P38, and JNK was assessed through Western blot. PAS staining showed that PR decreased renal tubular injury in UUO mice. SR and IHC staining demonstrated that PR decreased the accumulation of ECM. PR treatment significantly inhibited epithelial cell apoptosis according to the results of TUNEL, FCM, Hoechst, and Western blot. Furthermore, NOX4 increased in UUO mice and decreased with PR treatment. HO-derived oxidative stress activated epithelial apoptosis and mitogen-activated protein kinases (MAPK), and PR treatment significantly reversed it. These results suggest that PR treatment ameliorates renal fibrosis by inhibiting oxidative stress induced-epithelial cell apoptosis through MAPK signaling.
葛根素（PR）是从植物野葛的根中分离出的一种异黄酮，已被广泛应用于传统中药中治疗各种疾病。氧化应激和上皮细胞凋亡在肾纤维化过程中起重要作用。本研究旨在确定PR是否通过减少氧化应激诱导的上皮细胞凋亡来抑制肾纤维化。在体内，单侧输尿管梗阻（UUO）诱导肾纤维化和上皮细胞凋亡。总共24只小鼠被随机分为四个实验组：假手术组、单纯UUO组、UUO + 50mg/kg PR组和UUO + 100mg/kg PR组。在体外，200μM过氧化氢（HO）诱导上皮细胞凋亡。实验分为四组：对照组、单纯HO组、HO + 50μM PR组和HO + 100μM PR组。通过过碘酸 - 希夫（PAS）染色测量小鼠肾皮质中的肾小管损伤，并通过天狼星红（SR）、免疫组织化学（IHC）染色和蛋白质印迹法测量细胞外基质（ECM）。通过末端脱氧核苷酸转移酶介导的dUTP缺口末端标记（TUNEL）、流式细胞术（FCM）和Hoechst检测来测量肾上皮细胞凋亡。通过蛋白质印迹法评估NOX4、caspase3、ERK、P38和JNK的蛋白质表达。PAS染色显示PR减少了UUO小鼠的肾小管损伤。SR和IHC染色表明PR减少了ECM的积累。根据TUNEL、FCM、Hoechst和蛋白质印迹法的结果，PR治疗显著抑制了上皮细胞凋亡。此外，NOX4在UUO小鼠中增加，并随着PR治疗而减少。HO衍生的氧化应激激活上皮细胞凋亡和丝裂原活化蛋白激酶（MAPK），而PR治疗显著逆转了这一过程。这些结果表明，PR治疗通过MAPK信号传导抑制氧化应激诱导的上皮细胞凋亡，从而改善肾纤维化。
