[1]陈李泽成,李金凤,乔锋△.调气汤对大鼠类风湿性关节炎的治疗作用和机制研究[J].中国中医骨伤科杂志,2023,31(02):6-11.
　CHEN-LI Zecheng LI Jinfeng QIAO Feng.Study on the Efficacy and Mechanism of Tiaoqi Decoction on the Treatment of Rheumatoid Arthritis in Rats[J].Chinese Journal of Traditional Medical Traumatology & Orthopedics,2023,31(02):6-11.
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调气汤对大鼠类风湿性关节炎的治疗作用和机制研究()
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《中国中医骨伤科杂志》[ISSN:1005-0205/CN:42-1340/R]

卷:
第31卷
期数:
2023年02期
页码:
6-11
栏目:
实验研究
出版日期:
2023-02-15

文章信息/Info

Title:
Study on the Efficacy and Mechanism of Tiaoqi Decoction on the Treatment of Rheumatoid Arthritis in Rats
文章编号:
1005-0205(2023)02-0006-06
作者:
陈李泽成1; 李金凤1; 乔锋1△
1陕西中医药大学第一临床医学院(陕西 咸阳,712046)
Author(s):
CHEN-LI Zecheng1 LI Jinfeng1 QIAO Feng1△
1The First Clinical Medical College of Shaanxi University of Traditional Chinese Medicine, Xianyang 712046, Shaanxi China.
关键词:
调气汤;  类风湿性关节炎;  成纤维滑膜细胞;  自噬;  肿瘤坏死因子
Keywords:
Tiaoqi decoction;  rheumatoid arthritis;  fibroblast synoviocytes;  autophagy;  TNF-α
分类号:
R684.3
文献标志码:
A
摘要:
目的:研究调气汤对类风湿性关节炎的治疗作用和机制。方法:体外培养胶原诱导型关节炎大鼠成纤维样滑膜(CIA-FLS)细胞,将细胞分为对照组、TNF-α组、调气汤40,50和60 μg/mL组,TNF-α(10 ng/mL)处理细胞30 min,然后用调气汤处理细胞48 h。检测细胞活力、细胞凋亡和细胞中Atg5、Beclin-1、LC3-Ⅰ/Ⅱ、PI3K、p-AKT、AKT、p-mTOR的蛋白表达,以及细胞上清中TNF-α、IL-6、TGF-β、IL-17、IFN-γ和IL-10含量。SPF级SD大鼠(雄性,30只)随机分为对照组、模型组、低剂量组(25 mg/kg调气汤的佐剂诱导型关节炎(AIA)大鼠)、中剂量组(50 mg/kg调气汤的AIA大鼠)、高剂量组(100 mg/kg调气汤的AIA大鼠),建立胶原诱导型关节炎(CIA)大鼠模型。采用苏木精-伊红染色观察AIA大鼠的组织病理变化,采用ELISA法测量大鼠血清中TNF-α、IL-6、IL-1β含量。结果:与TNF-α组比较,调气汤各剂量组细胞活力下降、细胞凋亡增加,且细胞中Atg5、Beclin-1、LC3-Ⅰ/Ⅱ、PI3K、p-mTOR蛋白表达下调,同时细胞上清中TNF-α、IL-6、TGF-β、IL-17、IFN-γ含量减少,IL-10含量增加,差异有统计学意义(P<0.05)。与模型组比较,调气汤低剂量组的组织病理图像中观察到轻度骨侵蚀; 调气汤中剂量和高剂量组的组织病理图像显示为正常的关节腔,没有骨侵蚀和滑膜增生; 血清上清中TNF-α、IL-6和IL-1β含量均明显下降,差异有统计学意义(P<0.05)。结论:调气汤通过下调PI3K/AKT/mTOR信号通路抑制CIA-FLS细胞增殖和自噬,并促进细胞凋亡和发挥抗炎活性,对类风湿性关节炎具有潜在的治疗活性。
Abstract:
Objective:To investigate the effect and mechanism of Tiaoqi decoction on the treatment of rheumatoid arthritis. Methods:Collagen-induced arthritis fibroblast-like synovial(CIA-FLS)cells of collagen-induced arthritis rats were cultured in vitro. The cells were divided into control group, TNF-α group, Tiaoqi decoction 40 μg/mL, 50 μg/mL and 60 μg/mL groups. TNF-α(10 ng/mL)was used to treat the cells for 30 min, and then cells were treated with Tiaoqi decoction for 48 h. Cell viability, apoptosis, protein expression of Atg5, Beclin-1, LC3-Ⅰ/Ⅱ, PI3K, p-AKT, AKT, p-mTOR in cells was detected. The content of TNF-α, IL-6, TGF-β, IL-17, IFN-γ and IL-10 in the cell supernatant was measured. 30 male SPF SD rats were randomly divided into control group, model group, low-dose group(25 mg/kg Tiaoqi decoction adjuvant induced arthritis(AIA)rats), middle-dose group(50 mg/kg Tiaoqi decoction AIA rats), high-dose group(100 mg/kg Tiaoqi decoction AIA rats). A collagen-induced arthritis(CIA)rat model was established. Hematoxylin-eosin staining was used to observe the pathological changes of AIA rats, and ELISA was used to measure the contents of TNF-α, IL-6, and IL-1β in rat serum. Results:Compared with TNF-α group, the cell viability was decreased and the apoptosis was increased in Ttiaoqi decoction group. The protein expression of Atg5, Beclin-1, LC3-Ⅰ/Ⅱ, PI3K and p-mTOR was down-regulated by Tiaoqi decoction. The content of TNF-α, IL-6, TGF-β, IL-17 and IFN-γ was decreased and the content of IL-10 was increased in the cell supernatant of Tiaoqi decoction group. Compared with the model group, mild bone erosion was observed in the pathological images of the Tiaoqi decoction low-dose group, the histological study of the Tiaoqi decoction medium-dose and high-dose groups showed normal joint cavity without bone erosion and slippage, the levels of TNF-α, IL-6 and IL-1β in the serum supernatant were all significantly decreased, the difference was statistically significant(P<0.05). Conclusion:Tiaoqi decoction can inhibit the proliferation and autophagy of CIA-FLS cells by down-regulating PI3K/AKT/mTOR signaling pathway, and promote cell apoptosis and exert anti-inflammatory activity, which has potential therapeutic activity on rheumatoid arthritis.

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备注/Memo

备注/Memo:
△通信作者 E-mail:qiaofeng7502@163.com
更新日期/Last Update: 2023-02-10