黄芩苷对脂多糖诱导的大鼠滑膜RSC-364细胞自噬的抑制作用

doi:10.13481/j.1671-587x.20200311

吉林大学学报(医学版) ›› 2020, Vol. 46 ›› Issue (03): 498-503.doi: 10.13481/j.1671-587x.20200311

• 基础研究 • 上一篇

黄芩苷对脂多糖诱导的大鼠滑膜RSC-364细胞自噬的抑制作用

王映映¹, 孙利娟¹, 范紫微², 古虹², 游先梅³, 关天昊⁴, 张成义², 陈曦¹

1. 北华大学医学院病原学教研室, 吉林吉林 132013;
2. 北华大学药学院药理教研室, 吉林吉林 132013;
3. 北华大学医学院医学影像学教研室, 吉林吉林 132013;
4. 北华大学医学技术学院医学检验技术教研室, 吉林吉林 132013

收稿日期:2019-09-12 发布日期:2020-06-11
通讯作者: 张成义,教授,硕士研究生导师(0432-64608281,E-mail:zhchyjl@163.com);陈曦,教授,硕士研究生导师(0432-64608072,E-mail:846617750@qq.com) E-mail:zhchyjl@163.com;846617750@qq.com
作者简介:王映映(1991-),女,河南省周口市人,在读医学硕士,主要从事类风湿关节炎机制方面的研究。
基金资助:
吉林省科技厅科技攻关项目资助课题（20190304048YY）；吉林省吉林市科技局科技计划项目资助课题（201820847）；吉林省中医药管理局中医药科技项目资助课题（2018111）；北华大学研究生创新计划项目资助课题（2019068）

Inhibitory effect of baicalin on autophagy of synovial RSC-364 cells of rats induced by lipopolysaccharide

WANG Yingying¹, SUN Lijuan¹, FAN Ziwei², GU Hong², YOU Xianmei³, GUAN Tianhao⁴, ZHANG Chengyi², CHEN Xi¹

1. Department of Pathogenic Biology, College of Medical Sciences, Beihua University, Jilin 132013, China;
2. Department of Pharmacology, College of Pharmacy, Beihua University, Jilin 132013, China;
3. Department of Medical Imageology, College of Medical Sciences, Beihua University, Jilin 132013, China;
4. Department of Medical Laboratory Technology, College of Medical Technology, Beihua University, Jilin 132013, China

Received:2019-09-12 Published:2020-06-11

摘要/Abstract

摘要： 目的：探讨黄芩苷通过磷脂酰肌醇3-激酶/蛋白激酶B/雷帕霉素（PI3k/Akt/mTOR）信号通路对脂多糖（LPS）诱导的大鼠滑膜RSC-364细胞自噬的影响，并阐明其作用机制。方法：将对数生长期大鼠滑膜RSC-364细胞分为对照组、模型组、地塞米松（DXMS）组、10 μmol·L^-1黄芩苷组、20 μmol·L^-1黄芩苷组和40 μmol·L^-1黄芩苷组。对照组RSC-364细胞只加入培养基，其他各组RSC-364细胞均采用1 mg·L^-1LPS体外刺激12 h制作炎症细胞模型。采用MTT法检测各组RSC-364细胞存活率，RT-PCR法检测各组RSC-364细胞中PI3k、Akt、mTOR、Beclin1和微管相关蛋白-轻链3-Ⅱ（LC3-Ⅱ） mRNA表达水平，Western blotting法检测各组RSC-364细胞中Beclin1、Atg5、Atg7、Atg12、LC3-Ⅱ和P62蛋白表达水平。结果：与对照组比较，模型组RSC-364细胞存活率明显升高（P<0.01），模型组RSC-364细胞中PI3k、Akt、mTOR mRNA和P62蛋白表达水平明显降低（P<0.05或P<0.01），Atg5、Atg7、Atg12、LC3-Ⅱ、Beclin1 mRNA及蛋白表达水平明显升高（P<0.01）；与模型组比较，不同浓度黄芩苷组RSC-364细胞存活率明显降低（P<0.05或P<0.01），RSC-364细胞中PI3k、Akt、mTOR mRNA和P62蛋白表达水平明显升高（P<0.05或P<0.01），Atg5、Atg7、Atg12、LC3-Ⅱ、Beclin1 mRNA和蛋白表达水平降低（P<0.05或P<0.01）。结论：黄芩苷可能通过激活PI3k/Akt/mTOR信号通路抑制LPS诱导的RSC-364细胞自噬。

关键词: 黄芩苷, RSC-364细胞, 风湿性关节炎滑膜成纤维细胞, 自噬

Abstract: Objective: To investigate the effect of baicalin on the autophagy of the synovial RSC-364 cells of the rats induced by lipopolysaccharide (LPS)through PI3k/Akt/mTOR signal pathway, and to clarify its mechanism. Methods: The rat synovial RSC-364 cells in logarithmic growth phase were divided into control group, model group, dexamethasone(DXMS) group, 10μmol·L^-1baicalin group, 20μmol·L^-1 baicalin group and 40μmol·L^-1 baicalin group. The RSC-364 cells in control group were only supplemented with culture medium, and the RSC-364 cells in the other groups were stimulated with 1 mg·L^-1LPS for 12 h to make the inflammatory cell models. The survival rates of RSC-364 cells were detected by MTT assay, and the expression levels of PI3k, Akt, mTOR, Beclin1, and LC3-Ⅱ mRNA in the RSC-364 cells in various groups were detected by RT-PCR method;Western blotting method was used to detect the expression levels of Beclin1, Atg5, Atg7, Atg12, microtubule-associated protein-light chain 3-Ⅱ(LC3-Ⅱ), and P62 proteins in the RSC-364 cells in various groups. Results: Compared with control group, the survival rate of RSC-364 cells in model group was significantly increased (P<0.01), the expression levels of PI3k, Akt, mTOR mRNA and P62 protein in the RSC-364 cells in model group were significantly decreased (P<0.05 or P<0.01), and the expression levels of Atg5, Atg7, Atg12, LC3-Ⅱ, Beclin1 mRNA and proteins were significantly increased (P<0.01); compared with model group, the survival rates of RSC-364 cells in different concentrations of baicalin groups were significantly decreased (P<0.05 or P<0.01), the expression levels of PI3k, Akt, mTOR mRNA and P62 protein in the RSC-364 cells in different concentrations of baicalin groups were significantly increased (P<0.05 or P<0.01), and the expression levels of Atg5, Atg7, Atg12, LC3-Ⅱ and Beclin1 mRNA and proteins were decreased (P<0.05 or P<0.01). Conclusion: Baicalin may inhibit the LPS-induced autophagy of the RSC-364 cells by activating the PI3k/Akt/mTOR signaling pathway.

Key words: baicalin, RSC-364 cells, rheumatoid arthritis synovial fibroblasts, autophagy

中图分类号:

R329.28

王映映, 孙利娟, 范紫微, 古虹, 游先梅, 关天昊, 张成义, 陈曦. 黄芩苷对脂多糖诱导的大鼠滑膜RSC-364细胞自噬的抑制作用[J]. 吉林大学学报(医学版), 2020, 46(03): 498-503.

WANG Yingying, SUN Lijuan, FAN Ziwei, GU Hong, YOU Xianmei, GUAN Tianhao, ZHANG Chengyi, CHEN Xi. Inhibitory effect of baicalin on autophagy of synovial RSC-364 cells of rats induced by lipopolysaccharide[J]. Journal of Jilin University(Medicine Edition), 2020, 46(03): 498-503.

参考文献

[1] LEE W, KU S K, BAE J S. Anti-inflammatory effects of Baicalin, Baicalein, and Wogonin in vitro and in vivo[J].Inflammation,2015,38(1):110-125.
[2] WANG CC,SONG Y,WANG X,et al. Baicalin ameliorates collagen-induced arthritis through the suppression of Janus kinase 1(JAK1)/signal transducer and activator of transcription 3(STAT3) signaling in mice[J]. Med Sci Monit,2018,24(1):9213-9222.
[3] DING Y, DOU J, TENG Z,et al. Antiviral activity of baicalin against influenza A (H1N1/H3N₂) virus in cell culture and in mice and its inhibition of neuraminidase[J]. Arch Virol,2014,159(12):3269-3278.
[4] GAO C,ZHOU Y,LI H,et al. Antitumor effects of baicalin on ovarian cancer cells through induction of cell apoptosis and inhibition of cell migration in vitro[J]. Mol Med Rep,2017,16(6):8729-8434.
[5] WANG H Z,WANG H H,HUANG S S,et al. Inhibitory effect of baicalin on collagen-induced arthritis in rats through the nuclear factor-B pathway[J]. J Pharmacol Exp Ther,2014,350(2):435-443.
[6] YANG X,YANG J,ZOU H J.Baicalin inhibits IL-17-mediated joint inflammation in murine adjuvant-induced arthritis[J]. Clin Dev Immunol,2013,2013(1):268065.
[7] VALERIA M,SERENA R,ANTONELLA C,et al. Autophagy induces protein carbamylation in fibroblast-like synoviocytes from patients with rheumatoid arthritis[J]. Rheumatology(Oxford),2018,57(11):2032-2041.
[8] KATO M,OSPELT C,GAY R E,et al. Dual role of autophagy in stress-induced cell death in rheumatoid arthritis synovial fibroblasts[J]. Arthritis Rheumatol,2014,66(1):40-48.
[9] ZHU L,WANG H Z,WU Y, et al. The autophagy level is increased in the synovial tissues of patients with active rheumatoid arthritis and is correlated with disease severity[J]. Mediators Inflamm,2017,2017(6):7623145.
[10] 冯楠楠,王冰洁,朱启航,等. 玉米赤霉烯酮通过PI₃K/Akt/mTOR通路诱导大鼠睾丸支持细胞自噬及对细胞周期分布的影响[J]. 南京农业大学学报,2018,41(4):708-714.
[11] VOMERO M,MANGANELLI V,BARBATI C,et al. Reduction of autophagy and increase in apoptosis correlates with a favorable Clinical outcome in patients with Rheumatoid arthritis treated with anti-TNF drugs[J]. Arthritis Res Ther,2019,21(1):39.
[12] PEETERS J G C,De GRAEFF N,LOTZ M,et al. Increased autophagy contributes to the inflammatory phenotype of juvenile idiopathic arthritis synovial fluid T cells[J]. Rheumatology(Oxford),2017,56(10):1694-1699.
[13] TONG W W,ZHANG C,HONG T,et al. Silibinin alleviates inflammation and induces apoptosis in human rheumatoid arthritis fibroblast-like synoviocytes and has a therapeutic effect on arthritis in rats[J]. Sci Rep,2018,8(1):3241.
[14] MARINO G,NISO-SANTANO M,BAEHRECKE E H,et al. Self-consumption:the interplay of autophagy and apoptosis[J]. Nat Rev Mol Cell Biol,2014,15(2):81-94.
[15] XU K,CAI Y S,LU S M,et al. Autophagy induction contributes to the resistance to methotrexate treatment in rheumatoid arthritis fibroblast-like synovial cells through high mobility group box chromosomal protein 1[J]. Arthritis Res Ther,2015,17(1):374.
[16] RAYCHAUDHURI S K,RAYCHAUDHURI S P. mTOR signaling cascade in psoriatic disease:double kinase mTOR inhibitor a novel therapeutic target[J].Indian J Dermatol,2014,59(1):67-70.
[17] TANG J,LU X J, CHEN F F. Effects of perfluorooctanoic acid on the associated genes expression of autophagy signaling pathway of carassius auratus lymphocytes in vitro[J].Front Physiol,2018,9:1748.
[18] KIM E K,KWON J E,LEE S Y,et al. IL-17-mediated mitochondrial dysfunction impairs apoptosis in rheumatoid arthritis synovial fibroblasts through activation of autophagy[J]. Cell Death Dis,2017, 8(1):e2565.
[19] 洪永桃.自噬在类风湿关节炎滑膜成纤维细胞中的作用及甲氨蝶呤对自噬的影响[D].南充:川北医学院,2015.

黄芩苷对脂多糖诱导的大鼠滑膜RSC-364细胞自噬的抑制作用

Inhibitory effect of baicalin on autophagy of synovial RSC-364 cells of rats induced by lipopolysaccharide

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