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缪成贵, 熊友谊, 秦梅颂, 等. 芍药苷抑制佐剂性关节炎大鼠mTOR信号通路的实验研究[J]. koko体育app 学报（医学版）, 2018, 49(4): 535-539.
引用本文: 缪成贵, 熊深厚友谊, 秦梅颂, 等. 芍药苷阻止佐剂性膝盖炎大鼠mTOR警报环路的实验室探析[J]. 河北读书学报（生物学版）, 2018, 49(4): 535-539.
MIAO Cheng-gui, XIONG You-yi, QIN Mei-song, et al. Experimental Study on Paeoniflorin Inhibiting mTOR Signaling Pathway in Adjuvant Arthritis Rats[J]. Journal of Sichuan University (Medical Sciences), 2018, 49(4): 535-539.
Citation: MIAO Cheng-gui, XIONG You-yi, QIN Mei-song, et al. Experimental Study on Paeoniflorin Inhibiting mTOR Signaling Pathway in Adjuvant Arthritis Rats[J]. Journal of Sichuan University (Medical Sciences), 2018, 49(4): 535-539.

芍药苷抑制佐剂性关节炎大鼠mTOR信号通路的实验研究

Experimental Study on Paeoniflorin Inhibiting mTOR Signaling Pathway in Adjuvant Arthritis Rats

  • 摘要: 目的 研究芍药苷（paeoniflorin，PF）对佐剂性关节炎（adjuvant arthritis，AA）大鼠关节滑膜成纤维样滑膜细胞（fibroblast-like synoviocytes，FLS）内mTOR信号的影响。 方法 将大鼠分为正常组、AA组、AA+PF 100 μg/mL组、AA+PF 200 μg/mL组、AA+PF 400 μg/mL组和AA+PBS组，除正常组外，其余各组大鼠均采用完全弗氏佐剂足跖注射法制备AA大鼠，AA+PF 100 μg/mL组、AA+PF 200 μg/mL组、AA+PF 400 μg/mL组大鼠造模后向尾静脉注射剂量为0.1 mL/200g体重PF，研究3个剂量PF对AA大鼠关节炎评分的影响。造模后第28天股动脉放血处死正常组和AA组大鼠，分离各组大鼠关节滑膜组织，培养FLS，将培养出的FLS分为正常组、AA组、AA+PF 1 μg/mL、AA+PF 2 μg/mL、AA+PF 4 μg/mL组，通过Real time qPCR检测PF加药对AA大鼠FLS内mTOR表达的影响，Western blot检测PF对AA大鼠FLS磷酸化mTOR（p-mTOR）蛋白表达的影响，ELISA检测PF对AA大鼠FLS细胞因子白细胞介素（IL）-1、IL-6表达的影响，real time qPCR检测PF加药对AA大鼠FLS内MMP3表达的影响，向FLS转染mTOR过表达载体48 h后再次检测IL-1、IL-6和MMP3的表达。 结果 PF能够降低AA大鼠关节炎评分。PF加药后48 h，与AA组FLS相比，AA大鼠FLS在PF为1、2、4 μg/mL时mTOR表达均降低。与AA组FLS相比，AA大鼠FLS在PF为2 μg/mL时p-mTOR蛋白表达量也降低。PF加药后48 h，与AA组FLS相比，AA大鼠FLS在PF为1、2、4 μg/mL时IL-1、IL-6、MMP3表达均降低。向AA+PF 2 μg/mL组FLS转染mTOR过表达载体48 h，与转染前相比，IL-1、IL-6、MMP3表达均升高。 结论 PF可以改善AA大鼠病理，其机制可能与AA大鼠FLS内mTOR信号的抑制有关。  
    Abstract: Objective To study the effect of paeoniflorin (PF) on mTOR signal in synovial fibroblast-like synoviocytes (FLS) in rats with adjuvant arthritis. Methods AA model rats were prepared by complete Freund's adjuvant injection in foot-plantar, the PF was injected to rats in AA + PF 100 μg/mL group, AA + PF 200 μg/mL group and AA + PF 400 μg/mL group by the tail vein injection at the dose of 0.1 mL/200 g body mass, and the effects of three doses of PF on arthritis scores in AA rats were studied. The modeling rats and control rats were sacrificed at 28 d after modeling, then the synovium was separeated from rat articular, the FLS were cultured. The effect of PF on the expression of mTOR and MMP3 in AA FLS was detected by the real time qPCR. The effect on the cytokine IL-1, IL-6 was detected by ELISA, and the Western blot was used to investigate the role of PF in the mTOR phosphorylation. Furthermore, FLS were transfected with mTOR vectors, and the effect of mTOR overexpression on the PF roles was detected by real time qPCR and ELISA. Results The tail vein injection of PF can significantly reduce the AA rat arthritis score. Compared with AA group, the expression of mTOR in AA+PF 1 μg/mL, AA+PF 2 μg/mL, AA+PF 4 μg/mL was significantly decreased at 48 h after dosing. Compared with AA group, the relative expression of p-mTOR protein in PF 2 μg/mL group was also decreased. Compared with AA group at 48 h after dosing, the levels of IL-1, IL-6 and MMP3 in AA+PF 1 μg/mL, AA+PF 2 μg/mL, AA+PF 4 μg/mL were significantly decreased, respectively. Compared with PF 2 μg/mL group, the relative expression of IL-1, IL-6 and MMP3 in PF 2 μg/mL+mTOR vectors was increased. Conclusion PF can significantly inhibit the pathology of AA rats, and its mechanism may be related to the inhibition of mTOR signal in FLS of AA rats.  
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