摘要
目的基于NDRG2/JAK2/STAT3信号通路探讨祛寒逐风合剂对类风湿关节炎成纤维样滑膜细胞(RA-FLS)增殖和凋亡的影响及作用机制。方法体外培养RA-FLS,将细胞分为:(1)空白血清组、甲氨蝶呤组和祛寒逐风合剂低、中、高剂量组;(2)空白血清组、AG490组和祛寒逐风合剂低、中、高剂量组。采用不同浓度含药血清干预细胞,CCK-8法检测细胞增殖,流式细胞术检测细胞凋亡,RT-qPCR检测细胞Bax、Bcl-2、Caspace-3、Caspace-9、N-myc下游调控基因2(NDRG2)、Janus激酶2(JAK2)、信号转导和转录激活因子3(STAT3)m RNA表达,Western blot检测细胞Bax、Bcl-2、Caspace-3、Caspace-9、NDRG2、JAK2、STAT3、p-JAK2、p-STAT3蛋白表达。结果与空白血清组比较,甲氨蝶呤组和祛寒逐风合剂各剂量组细胞存活率明显降低(P<0.01),细胞凋亡率明显升高(P<0.01),Bax、Caspase-9 mRNA和蛋白表达明显升高(P<0.05,P<0.01),Bcl-2 mRNA和蛋白表达明显降低(P<0.01),甲氨蝶呤组和祛寒逐风合剂中、高剂量组Caspase-3 mRNA和蛋白表达明显升高(P<0.01);与空白血清组比较,祛寒逐风合剂各剂量组NDRG2 mRNA和蛋白表达明显升高(P<0.05,P<0.01),AG490组和祛寒逐风合剂中、高剂量组JAK2、STAT3 mRNA和蛋白表达明显降低(P<0.05,P<0.01),p-JAK2、p-STAT3蛋白表达明显降低(P<0.01)。结论祛寒逐风合剂可通过调节NDRG2/JAK2/STAT3信号通路活性调控RA-FLS增殖和凋亡,发挥治疗类风湿关节炎的作用。
Objective To explore the mechanism of Quhan Zhufeng Mixture on proliferation and apoptosis of rheumatoid arthritis fibroblast-like synoviocyte(RA-FLS)based on NDRG2/JAK2/STAT3 signaling pathway.Methods RA-FLS cells were cultured in vitro,and were divided into①blank serum group,methotrexate group,Quhan Zhufeng Mixture low-,medium-and high-dosage groups;②blank serum group,AG490 group,Quhan Zhufeng Mixture low-,medium-and high-dosage groups.Different concentrations of drug-containing serum were used to intervene cells.Cell proliferation was detected by CCK-8 method,apoptosis was detected by flow cytometry,and mRNA expressions of Bax,Bcl-2,Caspace-3,Caspace-9,N-myc downstream regulatory gene 2(NDRG2),Janus kinase 2(JAK2)and signal transduction and transcription activator 3(STAT3)were detected by RT-qPCR,Western blot was used to detect the protein expressions of Bax,Bcl-2,Caspace-3,Caspace-9,NDRG2,JAK2,STAT3,p-JAK2 and p-STAT3 in cells.Results Compared with the blank serum group,cell survival rate in methotrexate group,Quhan Zhufeng Mixture all dosage groups significantly decreased(P<0.01),the apoptosis rate significantly increased(P<0.01),the mRNA and protein expressions of Bax and Caspase-9 significantly increased(P<0.05,P<0.01),while the mRNA and protein expression of Bcl-2 significantly decreased(P<0.01),and Caspase-3 mRNA and protein expression in methotrexate group and Quhan Zhufeng Mixture medium-and high-dosage groups significantly increased(P<0.01).Compared with the blank serum group,the mRNA and protein expression of NDRG2 significantly increased in Quhan Zhufeng Mixture all dosage groups(P<0.05,P<0.01),the mRNA and protein expressions of JAK2 and STAT3 were significantly reduced in AG490 group and Quhan Zhufeng Mixture medium-and high-dosage groups(P<0.05,P<0.01),and the expressions of p-JAK2 and p-STAT3 proteins were significantly reduced(P<0.01).Conclusion Quhan Zhufeng Mixture can regulate the proliferation and apoptosis of RA-FLS by regulating the activity of NDRG2/JAK2/STAT3 signaling pathway,playing a role in treating rheumatoid arthritis.
作者
苏小军
朱文菊
郭影
王欢
何倩
张志明
田雪梅
沈海丽
马骏
包强
SU Xiaojun;ZHU Wenju;GUO Ying;WANG Huan;HE Qian;ZHANG Zhiming;TIAN Xuemei;SHEN Haili;MA Jun;BAO Qiang(Gansu Provincial Hospital of TCM,Lanzhou 730050,China;Gansu University of Chinese Medicine,Lanzhou 730030,China;The Second Hospital Lanzhou University,Lanzhou 730030,China)
出处
《中国中医药信息杂志》
2025年第7期119-126,共8页
Chinese Journal of Information on Traditional Chinese Medicine
基金
国家自然科学基金(82260880)
甘肃省自然科学基金(22JR5RA626)
甘肃省中医药管理局重点科研项目(GZKZ-2021-3)。
