中伦安神汤含药血清通过抑制 Janus 激酶/信号转导和转录激活因子信号通路诱导胶原诱导性关节炎大鼠成纤维样滑膜细胞凋亡的作用。
Efficacy of Zhonglun'a-decoction-containing serum on fibroblast-like synoviocyte apoptosis in rats with collagen-induced arthritis via inhibiting Janus kinase/signal transducer and activator of transcription signaling pathway.
机构信息
College of Traditional Chinese Medicine, Inner Mongolia Medical University, Hohhot 010000, China.
Rheumatology Department, Hohhot Hospital of Mongolian Medicine and Traditional Chinese Medicine, Hohhot 010010, China.
出版信息
J Tradit Chin Med. 2019 Apr;39(2):181-190.
OBJECTIVE
To investigate the efficacy of Zhonglun'a-decoction-containing serum (ZHONGL-CS) on the in vitro apoptosis of fibroblast-like synoviocytes (FLS) from rats with collagen-induced arthritis (CIA) by investigating the Janus kinase/signal transducer and activator of transcription (JAK/STAT) signaling pathway.
METHODS
A CIA rat model was established using bovine type Ⅱ collagen. FLS were isolated, cultured and identified. A cell counting kit-8 was used to detect cell activity. The half maximal inhibitory concentration (IC50) was calculated. Experimental subjects were divided into control, CIA, ZHONGL-CS, JAK2 inhibitor AG490, and ZHONGLCS with AG490 groups. The in vitro cell cycle and apoptosis rate were detected in FLS by flow cytometry. Bcl-2-associated X protein (Bax), B-cell lymphoma 2 (Bcl-2), caspase-3, cyclin D1, phosphorylated JAK2, STAT1, and STAT3 protein expressions in FLS were examined by Western blotting. JAK2, STAT1 and STAT3 mRNA levels were examined by quantitative real-time polymerase chain reaction.
RESULTS
Compared with the CIA group, FLS proliferation was inhibited, the FLS G0/G1 cell cycle was arrested, and the rate of FLS apoptosis was increased in the ZHONGL-CS group. In the ZHONGLCS group, the protein levels of Bcl-2 and cyclin D1 were reduced compared with the CIA group and the levels of Bax and caspase-3 in FLS were increased. In the ZHONGL-CS group, the expressions of JAK2, STAT1, and STAT3 mRNA and the levels of phosphorylated JAK2, STAT1, and STAT3 proteins were reduced.
CONCLUSION
ZHONGL-CS may induce FLS apoptosis in CIA rats. Activation of the JAK/STAT signaling pathway was inhibited in FLS in vitro.
目的
通过研究中伦汤（ZHONGL-CS）含药血清对胶原诱导关节炎（CIA）大鼠成纤维样滑膜细胞（FLS）体外凋亡的影响，探讨 Janus 激酶/信号转导和转录激活因子（JAK/STAT）信号通路。
方法
采用牛Ⅱ型胶原建立 CIA 大鼠模型。分离、培养和鉴定 FLS。用细胞计数试剂盒-8 检测细胞活性，计算半数抑制浓度（IC50）。将实验对象分为对照组、CIA 组、ZHONGL-CS 组、JAK2 抑制剂 AG490 组和 ZHONGLCS+AG490 组。通过流式细胞术检测 FLS 体外细胞周期和凋亡率。Western blot 检测 FLS 中 B 细胞淋巴瘤 2 相关 X 蛋白（Bax）、B 细胞淋巴瘤 2（Bcl-2）、半胱氨酸天冬氨酸蛋白酶-3（caspase-3）、细胞周期蛋白 D1、磷酸化 JAK2、STAT1 和 STAT3 蛋白的表达。用实时定量聚合酶链反应检测 JAK2、STAT1 和 STAT3mRNA 的水平。
结果
与 CIA 组相比，ZHONGL-CS 组可抑制 FLS 增殖，阻滞 FLS G0/G1 细胞周期，增加 FLS 凋亡率。与 CIA 组相比，ZHONGLCS 组 FLS 中 Bcl-2 和细胞周期蛋白 D1 蛋白水平降低，Bax 和 caspase-3 蛋白水平升高。ZHONGL-CS 组 JAK2、STAT1 和 STAT3mRNA 的表达及磷酸化 JAK2、STAT1 和 STAT3 蛋白水平降低。
结论
ZHONGL-CS 可能通过抑制 JAK/STAT 信号通路的激活，诱导 CIA 大鼠 FLS 凋亡。
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