买朱尼对兔子关节炎性软骨细胞凋亡的调控作用

doi:10.3969/j.issn.1671-7171.2019.02.003

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摘要目的探讨买朱尼对兔子关节炎性软骨细胞凋亡的调控作用。方法兔子软骨细胞添加白介素1β(IL-1β)模拟关节炎性软骨细胞,用制备的低、中和高剂量买朱尼及对照含药血清进行干预。MTT法检测细胞活力;Annexin V-FITC/PI双染色流式细胞术检测细胞凋亡;实时定量PCR和蛋白质印迹法检测Fas/FasL信号通路中死亡受体(Fas)、死亡配体(FasL)、含半胱氨酸的天冬氨酸蛋白水解酶8(caspase-8)、含半胱氨酸的天冬氨酸蛋白水解酶3(caspase-3)、Fas相关的死亡结构域白介素-1β-转换酶抑制蛋白(I-FLICE)和DNA修复酶(PARP)相关mRNA和蛋白表达水平。结果与模型组相比,买朱尼各剂量组显著减少IL-1β诱导的细胞活力下降(P<0.001),显著减少软骨细胞的早期凋亡(P<0.05),显著减少Fas、FasL、caspase-8和caspase-3的mRNA和蛋白表达(P<0.001),增加I-FLICE和PARP的mRNA和蛋白表达(P<0.001)。结论买朱尼·阿扎拉克通过抑制Fas/FasL信号通路减少IL-1β诱导的兔子软骨细胞凋亡。

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关键词 ：骨关节炎/胚胎学, 软骨细胞, 细胞凋亡, 药用制剂

Abstract：Objective To explore the regulation effect and mechanism of Maizhuni on the apoptosis of rabbit chondrocytes of arthritis. Methods Apoptosis of arthritic chondrocytes was induced by IL-1beta. Normal serum or low, medium and high dose of Maizhuni serum were added to culture medium to regulate apoptosis of arthritic chondrocytes. Cell viability was measured by MTT assay. Cell apoptosis was detected by flow cytometry using the Annexin V-FITC/PI double staining. The expression level of Fas, FasL, caspase-8, caspase-3, I-FLICE, PARP was analyzed by real time quantitative PCR and western blotting. Results Compared to normal serum mimic group, all doses of maizhuni-treated groups significantly increased the viability of chondrocytes (P<0.001) and significantly reduced early apoptosis of chondrocytes (P<0.05). Meanwhile, maizhuni significantly decreased the expression levels (mRNA and protein) of Fas, FasL, caspase-8 and caspase-3, while increased the expression of I-FLICE and PARP (P<0.001). Conclusion Maizhuni inhibits the apoptosis of rabbit chondrocytes induced by IL-1 beta through mechanism of regulating Fas/FasL signal pathway.

Key words： Osteoarthritis/EM Chondrocytes Apoptosis Pharmaceutical Preparations

收稿日期: 2018-02-05

PACS:

R684.3

基金资助:新疆维吾尔自治区自治区自然基金项目(编号:2015211C148)

通讯作者: E-mail:lvgangxj@163.com

引用本文:

苗德胜, 王筠, 程国杰, 吕刚, 杨春, 杨新军. 买朱尼对兔子关节炎性软骨细胞凋亡的调控作用[J]. 医学临床研究, 2019, 36(2): 217-223.
MIAO De-sheng, WANG Yun, CHENG Guo-jie , et al. Regulatory Effect of Maizhuni on Rabbit Arthritic Chondrocyte Apoptosis. JOURNAL OF CLINICAL RESEARCH, 2019, 36(2): 217-223.

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http://journal07.magtech.org.cn/yxlcyj/CN/10.3969/j.issn.1671-7171.2019.02.003 或 http://journal07.magtech.org.cn/yxlcyj/CN/Y2019/V36/I2/217