中医药导报2025，Vol.31Issue(4)：42-47,52,7.DOI:10.13862/j.cn43-1446/r.2025.04.007
肾复康调控细胞衰老抑制D-Gal诱导的HK-2细胞纤维化作用及机制
Shenfukang(肾复康)Inhibited the D-Gal-Induced HK-2 Fibrosis via Regulating Cell Senescence
摘要
Abstract
Objective:To investigate the intervention effect and mechanisms of Shenfukang on D-Galactose(D-Gal)-induced fibrosis model in HK-2 cells.Methods:D-Gal was used to establish the fibrosis model in HK-2 cells.The MTT assay was employed to evaluate the influence of Shenfukang on the viability of HK-2 cells under D-Gal culture conditions.HK-2 cells were divided into control group,D-Gal model group,Vitamin E groups(200 μg/mL),Shenfukang low-dose group(200 μg/mL)and Shenfukang high-dose group(400 μg/mL)with the intervention time of 24 h.Western blotting was utilized to detect the expression of α-smooth muscle actin(α-SMA),Collagen I,Janus kinase 2(JAK2),and signal transducers and activators of transcription 3(STAT3).Immunofluorescence was conducted to examine Ki-67 expression.β-galactosidase staining was performed to assess cell senescence.Real-time quantitative PCR(RT-PCR)was used to measure the expression levels of interleukin-1β(IL-1β)mRNA,IL-6 mRNA and IL-8 mRNA in cells.Enzyme-linked immunosorbent assay(ELISA)was employed to measure the levels of IL-1β,IL-6 and IL-8 in cell supernatants.Results:The D-Gal model group showed lower cell viability than control group(P<0.01).The cell viability in the Shenfukang 200 and 400 μg/mL groups was higher than the D-Gal model group(P<0.01).Then doses of 200 μg/mL and 400 μg/mL were chosen as the Shenfukang low and high doses,respectively.The relative expression levels of α-SMA and Collagen I proteins in the D-Gal model group HK-2 cells were higher than the control group(P<0.01).The vitamin E group,Shenfukang low-dose group and Shenfukang high-dose group showed lower α-SMA and Collagen I protein relative expression levels than D-Gal model group(P<0.01).The D-Gal model group showed lower relative fluorescent expression level of Ki-67 protein than control group(P<0.01),while higher β-galactosidase positivity rate than control group(P<0.01).The vitamin E group,Shenfukang low-dose group and Shenfukang high-dose group showed higher relative fluorescent expression levels of Ki-67 protein than D-Gal model group(P<0.05 or P<0.01),while lower β-galactosidase positivity rate than D-Gal model group(P<0.01).The D-Gal model group showed higher relative expression levels of IL-1β mRNA,IL-6 mRNA and IL-8 mRNA than control group(P<0.01).The vitamin E group,Shenfukang low-dose group and Shenfukang high-dose group showed lower relative expression levels of IL-1β mRNA,IL-6 mRNA and IL-8 mRNA than D-Gal model group(P<0.01).The D-Gal model group showed higher levels of IL-1β,IL-6,and IL-8 in the cell supernatants,than control group(P<0.01).The vitamin E group,Shenfukang low-dose group and Shenfukang high-dose group showed lower levels of IL-1β,IL-6,and IL-8 in the cell supernatants,than D-Gal model group(P<0.01).The D-Gal model group showed higher relative expression levels of JAK2 protein and p-STAT3/STAT3 than control group(P<0.01).The vitamin E group,Shenfukang low-dose group and Shenfukang high-dose group showed lower relative expression levels of JAK2 protein and p-STAT3/STAT3 than D-Gal model group(P<0.01).Conclusion:Shenfukang may inhibit the D-Gal-induced fibrosis model in HK-2 cells by regulating cellular senescence.关键词
肾纤维化/肾复康/细胞衰老/D-半乳糖/α-平滑肌肌动蛋白/β-半乳糖苷酶/Janus激酶2信号转导-转录活化蛋白3信号Key words
renal fibrosis/Shenfukang/cellular senescence/D-Galactose/α-smooth muscle actin/β-galac-tosidase/JAK2-STAT3 signaling分类
医药卫生引用本文复制引用
黄上,胡梁,郑燕姣,刘劲松,龚雅琪,黄睿,任佳乐..肾复康调控细胞衰老抑制D-Gal诱导的HK-2细胞纤维化作用及机制[J].中医药导报,2025,31(4):42-47,52,7.基金项目
湖南省自然科学基金面上项目(2023JJ30363) （2023JJ30363）
湖南省中医药研究院科研课题重点项目(202015) （202015）
湖南省中医药科研计划项目重点项目(2021202) （2021202）
湖南省科技厅-临床医疗技术创新引导项目(2021SK51004) （2021SK51004）
湖南中医药大学研究生创新课题(2023CX112) （2023CX112）
