摘要
目的观察丹参酮ⅡA配伍γ-分泌酶抑制剂3,5-二氟苯乙酰-L-丙氨酰-S-苯基甘氨酸-t-丁酯(DAPT)对转化生长因子-β1(TGF-β1)诱导的人源性肾小管上皮细胞HK-2细胞中纤维化标志物fibronectin(FN)、N-cadherin及Notch1/Jagged1信号通路分子Notch1、Jagged1表达的影响,探讨其防治肾纤维化的机制。方法?HK-2细胞分为5组:正常组(无特殊处理)、模型组(TGF-β110 ng/mL)、丹参酮ⅡA组(TGF-β110 ng/mL+丹参酮ⅡA 5μmol/L)、DAPT组(TGF-β110 ng/mL+DAPT 10μmol/L)、丹参酮ⅡA+DAPT组(TGF-β110 ng/mL+丹参酮ⅡA 5μmol/L+DAPT 10μmol/L)。采用RT-qPCR、Western Bolt及免疫荧光技术检测各组细胞间质纤维化指标FN、N-cadherin及Notch1、Jagged1mRNA及蛋白的表达。结果?正常组HK-2细胞FN、N-cadherin、Notch1、Jagged1 mRNA及蛋白呈低表达,模型组HK-2细胞FN、N-cadherin、Notch1、Jagged1 mRNA及蛋白表达较正常组升高(P<0.05),各干预组HK-2细胞FN、N-cadherin、Notch1、Jagged1 mRNA及蛋白表达较模型组降低(P<0.05)。丹参酮ⅡA+DAPT组HK-2细胞FN、N-cadherin、Jagged1 mRNA及蛋白表达低于丹参酮ⅡA组及DAPT组(P<0.05);丹参酮ⅡA+DAPT组HK-2细胞Notch1 mRNA及蛋白表达低于丹参酮ⅡA组(P<0.05)。结论?丹参酮ⅡA配伍DAPT用药可抑制TGF-β1诱导的HK-2细胞FN、N-cadherin的表达缓解肾间质纤维化,其机制可能与其协同干预Notch1/Jagged1信号通路传导有关。
Objective To study the effects of Tanshinone ⅡA combined with DAPT on the expression of fibrosis markers fibronectin(FN),N-cadherin and Notch1/Jagged1 signaling pathway molecules Notch1 and Jagged1 in human renal tubular epithelial cells(HK-2) induced by transforming growth factor-β1(TGF-β1),and to explore its mechanism for preventing and treating renal fibrosis.Methods The experiment cells were divided into five groups,ie,normal group(no treatment),model group(TGF-β1 10 ng/mL),Tanshinone Ⅱ A group(TGF-β110 ng/mL+Tanshinone Ⅱ A 5 μmol/L),DAPT group(TGF-β1 10 ng/mL+DAPT 10 μmol/L),TanshinoneⅡ A+DAPT group(TGF-β1 10 ng/mL+Tanshinone ⅡA 5 μmol/L+DAPT 10 μol/L).The expression of FN,N-cadherin and Notch1,Jagged1 mRNA and protein in each group were detected by RT-qPCR,Western Bolt and immunofluorescence.Results Normal HK-2 cells showed low expression of FN,N-cadherin,Notch1,Jagged1 mRNA and protein.The expressions of FN,N-cadherin,Notch1,Jagged1 mRNA and protein in the normal group were lower than those in the model group(P<0.05),and the expressions of FN,N-cadherin,Notch1,Jagged1 mRNA and protein in HK-2 cells in each intervention group were lower than those in the model group(P<0.05).The expressions of FN,N-cadherin,Jagged1 mRNA and protein in Tanshinone ⅡA+DAPT group were significantly lower than those in Tanshinone Ⅱ A group and DAPT group(P<0.05);the relative expressions of Notch1 mRNA and protein in Tanshinone Ⅱ A+DAPT group were lower than those in Tanshinone Ⅱ A group(P<0.05).Conclusions Tanshinone Ⅱ A combined with DAPT can inhibit the expression of FN and N-cadherin in HK-2 cells induced by TGF-β1 and relieve renal interstitial fibrosis.The mechanism might be correlated to its synergy effect on Notch1/Jagged1 signal pathways.
作者
李均
陈莎莎
LI Jun;CHEN Sha-sha(Key Laboratory of Applied and Basic Research of Traditional Chinese Medicine,Zhuhai Campus of Zunyi Medical University,Guangdong 519000)
出处
《中国中西医结合杂志》
CAS
CSCD
北大核心
2021年第9期1087-1092,共6页
Chinese Journal of Integrated Traditional and Western Medicine
基金
国家自然科学基金资助项目(No.81660746)。
