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万磊, 刘健, 黄传兵等. 雷公藤甲素调节佐剂关节炎大鼠滑膜、脾脏、胸腺组织细胞自噬的实验研究[J]. koko体育app 学报(医学版), 2017, 48(4): 520-525.
引用本文: 万磊, 刘健, 黄传兵等. 雷公藤甲素可以调节佐剂髋骨关节炎大鼠滑膜、脾脏、胸腺团体细胞核自噬的调查理论研究[J]. 重庆高校学报(医学检验版), 2017, 48(4): 520-525.
WAN Lei, LIU Jian, HUANG Chuanbing. et al. Effects of Triptolide on the Autophagy in Synovial, Spleen and Thymus of Rats with Adjuvant Arthritis[J]. Journal of Sichuan University (Medical Sciences), 2017, 48(4): 520-525.
Citation: WAN Lei, LIU Jian, HUANG Chuanbing. et al. Effects of Triptolide on the Autophagy in Synovial, Spleen and Thymus of Rats with Adjuvant Arthritis[J]. Journal of Sichuan University (Medical Sciences), 2017, 48(4): 520-525.

雷公藤甲素调节佐剂关节炎大鼠滑膜、脾脏、胸腺组织细胞自噬的实验研究

Effects of Triptolide on the Autophagy in Synovial, Spleen and Thymus of Rats with Adjuvant Arthritis

  • 摘要: 目的观察雷公藤甲素(TP)对佐剂关节炎(AA)大鼠滑膜脾脏胸腺自噬相关基因(Atg)/自噬标记蛋白(LC3-Ⅱ)、Beclin1表达和血清细胞因子水平的影响。方法将大鼠随机分为4组:正常对照 (NC) 组,模型对照 (MC) 组,来氟米特(LEF)组,雷公藤甲素(TP)组,每组12只,后3组复制成AA大鼠模型。致炎后第13 d开始给药。NC组、MC组给予生理盐水灌胃,每天1 次;LEF组按5 mg/kg的剂量灌胃,每天1次;TP组按50 μg/kg的剂量,每天1次。连续给药30 d。观察大鼠关节及其病理形态学变化,ELISA法检测血清细胞因子B淋巴细胞刺激因子(BAFF)、白介素(IL)-1、肿瘤坏死因子(TNF)-α、IL- 15、IL-10表达,RT-PCR法检测大鼠滑膜、脾脏、胸腺组织 Atg5 Atg7 Atg12 mRNA,Western blot法检测大鼠滑膜、脾脏、胸腺组织LC3-Ⅱ、Beclin1蛋白表达。结果 治疗后,TP组和LEP组大鼠足跖肿胀度(E)和关节炎指数较MC组降低。与NC组比较,MC组大鼠血清BAFF、IL-1、TNF-α升高,IL-15、IL-10降低,滑膜 Atg5 Atg12mRNA降低,脾脏 Atg5mRNA降低,脾脏 Atg7 Atg12mRNA及胸腺 Atg12mRNA升高,滑膜、脾脏、胸腺组织LC3-Ⅱ、Beclin1下降(P<0.05或0.01)。与MC组比较,TP组滑膜 Atg7 Atg12 mRNA降低,脾脏 Atg5 Atg7 Atg12 mRNA及胸腺中 Atg5 Atg7 mRNA降低, Atg12 mRNA升高;滑膜、脾脏、胸腺组织LC3-Ⅱ、Beclin 1升高(P<0.05)。与LEF组比较,TP组TNF-α、BAFF降低,E、IL-15升高(P<0.05或0.01)。TP组滑膜 Atg7mRNA及胸腺 Atg5 Atg7 mRNA表达降低,胸腺 Atg12mRNA及脾脏 Atg5 Atg7 Atg12mRNA表达升高。结论TP通过调节滑膜、胸腺、脾脏组织细胞自噬,改善关节滑膜炎症反应。  
    Abstract: Objective To determine the effect of triptolide (TP) on the expression of ATG /LC3-Ⅱ Beclin1 in synovial, spleen, and thymusof rats with adjuvant arthritis (AA). Methods Rats were divided for four groups: normal control (NC), model control (MC), leflunomide (LEF) treatment, and triptolide (TP)treatment, with 12 rats in each group.The AA model was established through Freund’s complete adjuvant (0.1 mL each) injection into the right foot plantar skin to introduce inflammation and 10 days of tail root injection of 0.05 mL Freund’s complete adjuvant for immunity strengthening. Drug administration started 13 days after induction of inflammation. Rats in the NC and MC groups were given normal saline (1 mL/100 g) once a day for 30 days, compared with 5 mg/kg of oral LEF for the rats in the LEF group and 50 μg/kg of oral TP for the rats in the TP group. Paw swelling (E), joint arthritis index(AI) and joint pathological changes of the rats were recorded. The serum expressions of cytokines B lymphocyte stimulating factor (BAFF), interleukin (IL)-1,tumor necrosis factor (TNF) alpha,IL-15, and IL-10were detected by ELISA. The expressions of Atg5,Atg7, and Atg12 mRNA in synovial, spleen, and thymus of the rats were detected by RT-PCR.The expressions of LC3-Ⅱ and Beclin1in synovial,spleen,and thymus of the rats were detected by Western blot assay. Results The AA model rats had lower serum BAFF, IL-1, TNF alpha, IL-15, and IL-10; lower Atg5and Atg12 mRNA in synovial; lower Atg5 mRNA,Atg7, and Atg12 mRNA in spleen; higher Atg12 mRNA in thymus; and lower LC3-Ⅱ and Beclin1 in synovial, spleen and thymus(P<0.05 or 0.01). TP treatment led to reduced paw swelling and arthritis index; declined Atg7 and Atg12 mRNA in synovial; declined Atg5, Atg7 mRNA and Atg12 mRNA in spleen; decreased Atg5 and Atg7mRNA in thymus; increased Atg12mRNA in thymus; and increased LC3-Ⅱ and Beclin1 in synovial, spleen and thymus (P<0.05 or 0.01). Compared with rats treated with LEF, TP treated rats had lower TNF-α and BAFF and higher E and IL-15 (P<0.05 or 0.01); as well as decreased expressions of Atg7 mRNA (synovial) and Atg5, Atg7 mRNA (thymus), and increased expressions of Atg12 mRNA (thymus) and Atg5,Atg7,Atg12 mRNA (spleen). Conclusion TP regulates autophagy in synovial, thymus and spleen of AA rats, and improves theirjointinflammatory response.   
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