论文部分内容阅读
目的:研究京尼平苷酸(GA)对佐剂性关节炎(AA)大鼠抗炎作用,探讨GA对AA大鼠滑膜细胞增殖影响的可能凋亡机制。方法:Wistar大鼠随机分为6组,分别为正常组,模型组,200,100,50 mg.kg-1GA组和0.75 mg.kg-1甲氨蝶呤(MTX)组,右后足趾皮内注射弗氏完全佐剂后第19天给药,治疗4周,测定足肿胀指标和血清肿瘤坏死因子-α(TNF-α),白介素-1β(IL-1β)水平。培养AA大鼠滑膜细胞给予GA(1,2,4μmol.L-1)和4μmol.L-1MTX处理,MTT法检测细胞增殖,Ho-echst/PI双染观察细胞凋亡,流式细胞仪检测凋亡细胞率,RT-PCR法检测Bcl-2和Bax基因mRNA表达水平。结果:200,100mg.kg-1GA可以明显降低足肿胀度和关节炎指数(P<0.05或P<0.01),肿胀抑制率分别为25.4%,21.37%,并抑制血清中TNF-α,IL-1β水平(P<0.05),TNF-α抑制率为34.61%,28%,IL-1β抑制率为29.05%,21.65%。GA(1~4μmol.L-1)处理AA大鼠滑膜细胞5 d,明显抑制细胞增殖。流式细胞仪检测显示1,2,4μmol.L-1GA组凋亡率分别为15.8%,24.3%,40.7%(P<0.01)。RT-PCR结果显示,不同浓度GA明显抑制Bcl-2 mRNA表达,而促进Bax mRNA表达(P<0.05或P<0.01)。结论:京尼平苷酸可抑制AA大鼠继发性炎症,降低血清TNF-α,IL-1β水平,体外可抑制AA大鼠滑膜细胞增殖及诱导细胞凋亡,作用机制与其下调Bcl-2和上调Bax基因mRNA表达有关。
Objective: To study the anti-inflammatory effect of geniposidic acid (GA) on adjuvant arthritis (AA) rats and to explore the possible apoptosis mechanism of GA on the proliferation of synoviocytes in AA rats. Methods: Wistar rats were randomly divided into 6 groups: normal group, model group, 200,100,50 mg.kg-1GA group and 0.75 mg.kg-1 methotrexate (MTX) group. On the 19th day after Freund’s complete adjuvant injection, the rats were treated for 4 weeks. The indexes of paw edema and serum levels of tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) were measured. Cultured AA rat synoviocytes were treated with GA (1, 2, 4μmol.L-1) and 4μmol.L-1MTX, MTT assay was used to detect cell proliferation, Ho-echst / PI double staining was used to observe apoptosis, flow cytometry The apoptotic rate was detected by RT-PCR and the mRNA expression of Bcl-2 and Bax was detected by RT-PCR. Results: 200,100mg.kg-1GA significantly reduced the degree of paw swelling and arthritis (P <0.05 or P <0.01), and the inhibition rates of swelling were 25.4% and 21.37%, respectively. The levels of TNF-α and IL- (P <0.05). The inhibition rate of TNF-α was 34.61%, 28%. The inhibition rate of IL-1β was 29.05% and 21.65% respectively. GA (1 ~ 4μmol.L-1) treated AA rat synovial cells for 5 days, significantly inhibited cell proliferation. Flow cytometry showed that the apoptotic rates of 1, 2, 4μmol.L-1GA group were 15.8%, 24.3% and 40.7%, respectively (P <0.01). RT-PCR results showed that GA at different concentrations significantly inhibited the expression of Bcl-2 mRNA and promoted the expression of Bax mRNA (P <0.05 or P <0.01). CONCLUSION: Geniposidic acid can inhibit the secondary inflammation of AA rats, decrease the level of serum TNF-α and IL-1β, inhibit the proliferation and induce the apoptosis of synoviocytes in AA rats in vitro, and its mechanism may be related to the down-regulation of Bcl- 2 and up-regulate Bax gene mRNA expression.