文献类型：期刊文献

中文题名：归芪定年方通过调节JAK2/STAT通路活性延缓小鼠肾系膜细胞衰老

英文题名：Guiqi Dingnian Prescription Delays Senescence of Mouse Mesangial Cells by Regulating JAK2/STAT Pathway Activity

作者：孙文平[1];伍志伟[2,3];薛娜[2]

第一作者：孙文平

机构：[1]甘肃省肿瘤医院,兰州730000;[2]甘肃中医药大学,兰州730000;[3]甘肃省高校重大疾病分子医学与中医药防治研究重点实验室,兰州730000

第一机构：甘肃省肿瘤医院,兰州730000

年份：2021

卷号：27

期号：12

起止页码：67

中文期刊名：中国实验方剂学杂志

外文期刊名：Chinese Journal of Experimental Traditional Medical Formulae

收录：CSTPCD;;北大核心:【北大核心2020】；CSCD:【CSCD2021_2022】；

基金：甘肃省自然科学基金项目(1610RJZA064);甘肃省高校重大疾病分子医学与中医药防治研究重点实验室开放基金项目(FZYX14-1);甘肃省高等学校创新基金项目(2021-268)。

语种：中文

中文关键词：归芪定年方;Janus酪氨酸蛋白激酶2(JAK2)/信号转导与转录激活因子(STAT)信号通路;小鼠肾系膜细胞;D-半乳糖;衰老

外文关键词：Guiqi Dingnian prescription(GDP);Janus tyrosine kinase 2(JAK2)/signal transducer and activator of transcription(JAK2/STAT)signaling pathway;mouse mesangial cells;D-galactose(D-gal);senescence

摘要：目的:探讨归芪定年方(GDP)对D-半乳糖(D-gal)诱导致衰型小鼠肾系膜细胞中Janus酪氨酸蛋白激酶2(JAK2)/信号转导与转录激活因子(STAT)信号通路相关分子表达水平的影响。方法:以D-gal 10 g·L^(-1)诱导复制小鼠肾系膜细胞衰老模型,经GDP 40 mg·L-1水煎剂处理3 d后,衰老相关β-半乳糖苷酶(SA-β-gal)染色测定细胞衰老状态,流式细胞术检测细胞周期,细胞增殖与活性检测(CCK-8)法检测细胞存活率,实时荧光定量聚合酶链式反应(Real-time PCR)检测肿瘤坏死因子-α(TNF-α),白细胞介素-6(IL-6),核转录因子-κB(NF-κB)和白细胞介素-1α(IL-1α)m RNA表达水平,蛋白免疫印迹法(Western blot)测定细胞中JAK2/STAT信号通路相关分子STAT1,磷酸化STAT1(p-STAT1),STAT3,p-STAT3蛋白水平。结果:CCK-8结果显示GDP最佳药物质量浓度为40 mg·L^(-1)。与空白组比较,模型组SA-β-gal细胞阳性率显著升高(P<0.01),G0/G1期细胞百分数明显升高(P<0.05),G2/M期和S期细胞百分数显著降低(P<0.01),TNF-α,IL-6,NF-κB和IL-1αm RNA表达水平显著上调(P<0.01),STAT1,p-STAT1,STAT3和p-STAT3蛋白水平显著升高(P<0.01);与模型组比较,模型+GDP组SA-β-gal细胞阳性率明显降低(P<0.05),G0/G1期百分数明显降低(P<0.05),G2/M期和S期细胞百分数显著增加(P<0.01),TNF-α,IL-6,NF-κB和IL-1αm RNA表达水平明显下调(P<0.05),STAT1,p-STAT1,STAT3和p-STAT3蛋白水平明显降低(P<0.05)。结论:GDP可延缓小鼠肾系膜细胞衰老的进程,其作用机制可能与下调细胞JAK2/STAT通路相关因子的表达水平相关。
Objective:To investigate the effects of Guiqi Dingnian prescription (GDP) on the expression of related molecules in Janus tyrosine kinase 2(JAK2)/signal transducer and activator of transcription(JAK2/STAT)signaling pathway of D-galactose(D-gal)-induced senescent mesangial cells.Method:The senescent mouse mesangial cells induced by 10 g·L^(-1) D-gal were continuously treated with 40 mg·L^(-1) GDP for three days.The senescence of the treated cells was determined by senescence-associated(SA)-β-gal staining.The cell cycle was detected by flow cytometry.The cell viability was analyzed using the cell counting kit-8(CCK-8).The m RNA expression levels of tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),nuclear transcription factor-κB(NF-κB),and IL-1αwere detected by real-time polymerase chain reaction(Real-time PCR).The protein expression levels of STAT1,phosphorylated STAT1(p-STAT1),STAT3,and p-STAT3 in the JAK2/STAT signaling pathway were determined by Western blot.Result:CCK-8 results showed that the optimal concentration of GDP was 40 mg·L^(-1).Compared with the blank group,the positive rate of SA-β-gal in the model group was significantly higher(P<0.01),the percentage of cells in G0/G1 phase was significantly increased(P<0.05),the percentage of cells in G2/M and S phase was significantly decreased(P<0.01).The m RNA expression levels of TNF-α,IL-6,NF-κB and IL-1αwere significantly increased(P<0.01).Compared with the model group,the model+GDP group exhibited significantly decreased SA-β-gal-positive cells(P<0.05),reduced cells in the G0/G1 phase(P<0.05),increased cells in the G2/M and S phases(P<0.01),and down-regulated TNF-α,IL-6,NF-κB,and IL-1αm RNA expression(P<0.05)and STAT1,p-STAT1,STAT3,and p-STAT3 protein expression(P<0.05).Conclusion:GDP delays the senescence of mouse mesangial cells possibly by down-regulating the expression of related molecules in the JAK2/STAT pathway.