Effect of Danggui Niantongtang on Autophagy and Apoptosis of Rheumatoid Arthritis Fibroblast-like Synoviocyte

Qi-jin LU; Jia-yu LI; Yi-si CAI; Hong-xin LIN; Dan-dan FAN; Li-gang JIE; Li-xia YUAN

doi:10.13422/j.cnki.syfjx.20220236

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Effect of Danggui Niantongtang on Autophagy and Apoptosis of Rheumatoid Arthritis Fibroblast-like Synoviocyte

更新时间：2021-12-14

- Effect of Danggui Niantongtang on Autophagy and Apoptosis of Rheumatoid Arthritis Fibroblast-like Synoviocyte
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 28, Issue 2, Pages: 19-26(2022)
- 作者机构：
  
  1.南方医科大学中医药学院，广州 510515
  2.南方医科大学检验与生物技术学院，广州 510515
  3.南方医科大学珠江医院，广州 510260
  4.南方医科大学第五附属医院，广州 510920
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20220236
  CLC： R2-0;R33;R289;R593.22
- Published：20 January 2022，
  
  Published Online：23 November 2021，
  
  Received：10 August 2021，
- 稿件说明：
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陆麒瑾,李佳钰,蔡义思等.当归拈痛汤对类风湿关节炎成纤维样滑膜细胞自噬与凋亡的影响[J].中国实验方剂学杂志,2022,28(02):19-26.

LU Qi-jin,LI Jia-yu,CAI Yi-si,et al.Effect of Danggui Niantongtang on Autophagy and Apoptosis of Rheumatoid Arthritis Fibroblast-like Synoviocyte[J].Chinese Journal of Experimental Traditional Medical Formulae,2022,28(02):19-26.
陆麒瑾,李佳钰,蔡义思等.当归拈痛汤对类风湿关节炎成纤维样滑膜细胞自噬与凋亡的影响[J].中国实验方剂学杂志,2022,28(02):19-26. DOI： 10.13422/j.cnki.syfjx.20220236.

LU Qi-jin,LI Jia-yu,CAI Yi-si,et al.Effect of Danggui Niantongtang on Autophagy and Apoptosis of Rheumatoid Arthritis Fibroblast-like Synoviocyte[J].Chinese Journal of Experimental Traditional Medical Formulae,2022,28(02):19-26. DOI： 10.13422/j.cnki.syfjx.20220236.

摘要

目的

研究当归拈痛汤对类风湿关节炎（RA）成纤维样滑膜细胞自噬与凋亡的影响，进一步探讨当归拈痛汤防治RA的相关机制。

方法

收集临床RA患者的滑膜组织，分离出成纤维样滑膜细胞（FLS）并进行培养，将RA-FLS细胞分为4组，分别为空白血清组、当归拈痛汤含药血清低、中、高剂量组。通过划痕实验，transwell侵袭实验检测当归拈痛汤对RA-FLS细胞迁移、侵袭能力的影响；Hoechst 33342法检测RA-FLS细胞凋亡水平；单丹磺酰尸胺（MDC）染色检测细胞自噬水平；实时荧光定量聚合酶链式反应（Real-time PCR）和蛋白免疫印迹法（Western blot）检测B细胞淋巴瘤-2（Bcl-2），Bcl-2相关X蛋白（Bax），微管相关蛋白1轻链3（LC3），自噬关键分子酵母Atg6同系物1（Beclin1）的mRNA和蛋白表达水平。

结果

与正常血清组比较，当归拈痛汤含药血清低、中、高剂量组均能减缓划痕愈合（

0.01），并且能显著减少侵袭到下室的RA-FLS细胞的数目（

0.01）；Hoechst33342结果显示当归拈痛汤含药血清低、中、高可以不同程度促进RA-FLS细胞凋亡（

0.01）；MDC染色后，低、中、高当归拈痛汤含药血清组中自噬小体明显减少；与空白含药血清组比较，当归拈痛汤含药血清低、中、高剂量干预后，Bcl-2，LC3，Beclin1的mRNA和蛋白的表达水平显著下降（

0.01），Bax的mRNA和蛋白的表达水平显著升高（

0.01）。

结论

当归拈痛汤对RA成纤维样滑膜细胞增殖有明显抑制作用，其作用机制可能与抑制自噬、促进凋亡有关。

Abstract

Objective

To explore the effect of Danggui Niantongtang （DGNTT） on cell apoptosis and autophagy in rheumatoid arthritis （RA） fibroblast-like synoviocytes （FLS）.

Method

RA-FLS were isolated and cultured from the synovial tissue of RA patients. The cells were treated with 10% blank serum （blank control group）， 10% sera containing low， medium and high doses of DGNTT. Wound healing assa and cell invasion test were applied to observe the effect of RA-FLS invasion technique. The apoptosis and autophagy level of RA-FLS cells was detected by Hoechst33342 method and monodansylcadaverine （MDC） staining. The mRNA and protein expression levels of B cell lymphoma-2 （Bcl-2）， Bcl-2 associated X protein （Bax）， microtubule-associated protein 1 light chain 3 （LC3）， autophagy key molecular yeast Atg6 homolog 1 （Beclin1） were detected by Real-time fluorescence quantitative polymerase chain reaction（Real-time PCR）and Western blot.

Result

Compared with the blank control group，each dose of serum could slow down the wound healing and significantly Reduce the number of RA-FLS cells invading the lower chamber（

0.01），the mRNA and protein expression levels of Bcl-2，LC3，Beclin1 were significantly decreased（

0.01）， and Bax were significantly increased（

0.01）. Hoechst33342 results showed that low， medium and high doses DGNTT could promote RA-FLS cell apoptosis. After MDC staining，autophagosome in low， medium and high doses DGNTT decreased significantly（

0.01）.

Conclusion

Danggui Niantongtang can effectively inhibit the proliferation of fibroblast-like synoviocytes. Its mechanism may be related to promote apoptosis and inhibit autophagy of fibroblast-like synoviocytes.

关键词

当归拈痛汤类风湿关节炎成纤维样滑膜细胞自噬凋亡

Keywords

Danggui Niantongtangrheumatoid arthritisfibroblast-like synoviocytesautophagyapoptosis

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