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目的:观察膝骨关节炎(KOA)大鼠心肺功能的变化及新风胶囊(XFC)对其影响,探讨可能的分子机制。方法:按随机数字表将40只大鼠随机分为正常对照(NC)组,模型对照(MC)组,氨基葡萄糖对照(GS)组,XFC组,每组10只。除NC组外,采用关节腔注射木瓜蛋白酶和L-半胱氨酸方法复制成KOA大鼠模型,造模后第14天开始给药。NC组及MC组均给予生理盐水,剂量为0.01 g/kg,其余2组分别给予GS、XFC混悬液,剂量分别为0.098 g/kg、0.375 g/kg。采用超声诊断仪检测大鼠心功能,动物肺功能仪检测大鼠肺功能,ELISA法检测BTLA、HVEM、IL-17、IL-4、TGF-β1;流式细胞术检测大鼠外周血CD4+CD25+Treg、CD4+CD25+Foxp3+Treg的表达。结果:与NC组比较,MC组体质量、E、E/A、FVC、FEV1、FEF25、FEF50、FEF75、MMF、PEF,BTLA、HVEM、IL-4、CD4+CD25+Treg、CD4+CD25+Foxp3+Treg明显降低,TGF-β1、IL-17明显升高(P<0.01或P<0.05)。与MC组比较,各组大鼠体质量,心功能参数E峰、E/A,肺功能参数FEV1、FEF50、FEF75、PEF,BTLA、HVEM、IL-4、CD4+CD25+Treg、CD4+CD25+Foxp3+Treg明显升高,关节软骨Mankin评分、心系数、肺系数、TGF-β1、IL-17明显降低(P<0.01或P<0.05);与GS组相比,XFC组体质量、Treg升高最为明显(P<0.01或P<0.05)。结论:新风胶囊改善KOA大鼠关节软骨Mankin评分,改善其心肺功能,其机制可能是促进BTLA-HVEM负调共刺激信号作用,诱导Treg免疫耐受,上调IL-4表达,下调IL-17、TGF-β1表达,抑制异常免疫炎症反应。
OBJECTIVE: To observe the changes of cardiopulmonary function in KOA rats and the effect of Xinfeng capsule (XFC) on it, and to explore the possible molecular mechanisms. Methods: Forty rats were randomly divided into normal control (NC) group, model control (MC) group, glucosamine control (GS) group and XFC group according to random number table. Except NC group, rats were injected with papain and L-cysteine intra-articularly into KOA rat model. The rats in NC group and MC group were given normal saline at a dosage of 0.01 g / kg. The other two groups were given GS and XFC suspensions at dosages of 0.098 g / kg and 0.375 g / kg, respectively. The heart function of rat was detected by ultrasonic diagnostic apparatus, lung function of rats was tested by pulmonary function test, BTLA, HVEM, IL-17, IL-4 and TGF-β1 were measured by ELISA. Flow cytometry was used to detect the percentage of CD4 + CD25 + Treg, CD4 + CD25 + Foxp3 + Treg. RESULTS: Compared with NC group, the body mass, E, E / A, FVC, FEV1, FEF25, FEF50, FEF75, MMF, PEF, BTLA, HVEM, IL-4, CD4 + CD25 + Treg, CD4 + CD25 + Foxp3 + Treg was significantly decreased, while TGF-β1 and IL-17 were significantly increased (P <0.01 or P <0.05). Compared with the MC group, body weight, E function, E / A, FEV1, FEF50, FEF75, PEF, BTLA, HVEM, IL-4, CD4 + CD25 + Treg, CD4 + CD25 (P <0.01 or P <0.05). Compared with the GS group, the body mass, Treg, Treg and Foxp3 + Treg in the XFC group were significantly higher than those in the XFC group The most obvious increase (P <0.01 or P <0.05). Conclusion: Xinfeng Capsule can improve the Mankin score of articular cartilage of KOA rats and improve its cardiopulmonary function. The mechanism may be that it can promote BTAR-HVEM negative co-stimulatory signal, induce Treg immune tolerance, up-regulate IL-4 expression, TGF-β1 expression, inhibition of abnormal immune inflammation.