秦艽、威灵仙及其配伍对类风湿关节炎成纤维样滑膜细胞的抑制、促凋亡作用及对相关炎症因子的影响

目的研究秦艽、威灵仙及其配伍对类风湿关节炎成纤维样滑膜细胞(RA FLSs)的抑制、促凋亡作用及对相关炎症因子的影响。方法将购得的RA FLSs复苏后,通过光学显微鉴定和Vimentin免疫荧光鉴定并传代培养,将生长密度为80%～90%的细胞随机分为空白组、阳性药(甲氨蝶呤)组、秦艽组、威灵仙组、秦艽-威灵仙药对组5组。将中药水煎液制成质量浓度为8000μg/m L的含药培养基,将阳性药制成质量浓度为400μg/m L的含药培养基,各组分别给予相应含药培养基进行干预,以CCK-8法检测各组RA FLSs的增殖活性,以Annexin V-FITC法检测各组RA FLSs的凋亡情况,以ELISA法检测各组RA FLSs培养上清液中类风湿因子(RF)、C-反应蛋白(CRP)的含量。结果光学显微鉴定结果显示购得的RA FLSs形态符合文献报道,免疫荧光鉴定结果显示购得的RA FLSs为人源RA FLSs,且纯度大于99%; CCK-8法检测结果显示,各药物组对RA FLSs增殖均具有抑制作用,其中威灵仙组对RA FLSs的增殖抑制作用最强(P<0.01);Annexin V-FITC法检...

Effects of Gentianae macrophyllae Radix and Clematidis chinensis Osbeck along with their compatibility on the inhibition,apoptosis-promoting effects of rheumatoid arthritis fibroblast-like synoviocytes and their effect on related inflammatory factors

Objective To study the inhibitory and promoting apoptosis effect of Gentianae macrophyllae Radix and Clematidis chinensis Osbeck along with their compatibility on rheumatoid arthritis fibroblast-like synoviocytes(RA FLSs)and their effect on related inflammatory factors.Methods The purchased RA FLSs after recovery were identified by optical micro-scopy and Vimentin immunofluorescence technique and were subcultured,cells with a growth density of 80%to 90%were randomly divided into 5 groups,including blank group,positive medicine(methotrexate)group,Gentianae macrophyllae Radix group,Clematidis chinensis Osbeck group and Gentianae macrophyllae Radix and Clematidis chinensis Osbeck couplet medicinals group.The water decoction of Chinese materia medica was made into medicated medium with a mass concentration of 8000μg/m L,the positive medicine was made into400μg/m L medium,and each group was given the corresponding medicated medium for intervention.The proliferation activity of RA FLSs in each group was detected by CCK-8 method,the apoptosis of RA FLSs in each group was detected by Annexin V-FITC method,and the content of rheumatoid factor(RF)and C-reactive protein(CRP)in the culture supernatant of RA FLSs in each group was detected by ELISA method.Results Optical microscopy showed that the morphology of RA FLSs purchased was in accordance with the literature reports.Immunofluorescence qualitative results showed that RA FLSs purchased was human RA FLSs,and the purity was more than 99%.CCK-8 method showed that the proliferation of RA FLSs was inhibited in each medicine group,and the inhibition of RA FLSs was the strongest in Clematidis chinensis Osbeck group(P<0.01);the results of Annexin V-FITC method showed that the apoptosis rate of RA FLSs in each medicine group significantly increased compared with that of the blank group,and the difference was statistically significant(P<0.01),among them the apoptosis rate of RA FLSs in the positive medicine group was significantly higher than that in the other three groups,and the difference was statistically significant(P<0.05 or P<0.01).The results of ELISA showed that the content of RF and CRP in the culture supernatant of RA FLSs in each medication group all significantly decreased compared with that in the blank group,and the difference was statistically significant(P<0.01),among them,the content of RF in Clematidis chinensis Osbeck group was statistically lower than that in the other three groups,and the difference was statistically significant(P<0.01).Conclusion Single Gentianae macrophyllae Radix,single Clematidis chinensis Osbeck and Gentianae macrophyllae Radix and Clematidis chinensis Osbeck couplet medicinals can inhibit the proliferation activity of RA FLSs,promote apoptosis of RA FLSs and inhibit the expression of RF and CRP in the culture supernatant of RA FLSs.Among them,at the same mass concentration,single Clematidis chinensis Osbeck had the strongest inhibitory effect on cell proliferation and secretion of RF.