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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13d开始给药。NC组、MC组给予生理盐水灌胃,每天1次;LEF组按5mg/kg的剂量灌胃,每天1次;TP组按50μg/kg的剂量,每天1次。连续给药30d。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、Atg12mRNA,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、Atg12 mRNA降低,脾脏Atg5 mRNA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,Atg12mRNA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Atg7 mRNA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。
Objective To observe the effect of triptolide (TP) on the expression of atg / autophagy marker protein (LC3-Ⅱ), Beclin1 and serum cytokines in thymus of the synovial membrane of adjuvant arthritis (AA) rats . Methods The rats were randomly divided into 4 groups: normal control (NC) group, model control (MC) group, leflunomide (LEF) group and triptolide (TP) Copy to AA rat model. After the first post-inflammatory dose 13d. The rats in NC group and MC group were given normal saline intragastrically once a day. LEF group was given intragastrically at a dose of 5mg / kg once a day, while in TP group at 50μg / kg once a day. Continuous administration 30d. The joint and pathological changes of the rats were observed. The expressions of BAFF, IL-1, TNF-α, IL-15 and IL-10 were detected by ELISA The expressions of Atg5, Atg7 and Atg12 mRNA in synovium, spleen and thymus of rats were detected by RT-PCR. The expression of LC3-Ⅱ and Beclin1 protein in synovium, spleen and thymus were detected by Western blot. Results After treatment, the degree of paw swelling (E) and arthritis index in TP group and LEP group were lower than those in MC group. Compared with NC group, serum BAFF, IL-1, TNF-α, IL-15 and IL-10 were increased, Atg5 and Atg12 mRNA in the MC group were decreased, Atg7 mRNA in the spleen, Atg7, Atg12mRNA increased, synovium, spleen, thymus LC3-Ⅱ, Beclin1 decreased (P <0.05 or 0.01). Compared with MC group, the expression of Atg7 and Atg12 mRNA in synovial membrane of TP group decreased, the expression of Atg5, Atg7, Atg12 mRNA and the expression of Atg5 and Atg7 in spleen, and Atg12 mRNA increased in synovial membrane of TP group, while the expression of LC3-Ⅱ and Beclin 1 in synovium, spleen and thymus (P <0.05). Compared with LEF group, the levels of TNF-α, BAFF and E, IL-15 in TP group decreased (P <0.05 or 0.01). The expression of Atg7 mRNA and Atg5, Atg7 mRNA in the synovial membrane of TP group decreased, the expression of Atg5 mRNA, Atg5, Atg7 and Atg12 mRNA in the thymus increased. Conclusion TP can improve synovial inflammation by regulating autophagy of synovial membrane, thymus and spleen.