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目的:探讨桂枝芍药知母汤(GT)对尿酸钠致痛风性关节炎(GA)模型大鼠关节滑膜组织中炎性信号表达的影响。方法:180只雄性SD大鼠随机分配到3个实验,分别为关节滑膜免疫组化实验,酶联免疫吸附测定(ELISA)实验,蛋白质免疫印迹(Western blot)实验。各实验取大鼠60只,按体重随机分为6组,每组10只,分别为模型组,正常组,GT高、中、低剂量组(4,8,16 g·kg~(-1)),秋水仙碱阳性药组(3×10-4g·kg~(-1))。实验组均ig给药,正常组、模型组给予等容积的蒸馏水,每天1次,连续给药7 d。第5天ig前,大鼠足踝关节注射尿酸钠悬液诱导GA。取大鼠关节滑膜组织,免疫组化检测Nod样受体蛋白3(NLRP3)炎性体的表达,Image-Pro Plus6.0图像分析系统测定平均积分吸光度(IA),Western blot检测凋亡相关斑点样蛋白(ASC),半胱氨酸天冬氨酸酶~(-1)(Caspase~(-1))信号衔接蛋白表达,ELISA测定炎性因子白细胞介素~(-1)β(IL~(-1)β),白细胞介素-6(IL-6),肿瘤坏死因子-α(TNF-α),核因子-κB(NF-κB)表达水平。结果:造模72 h后,与正常组比较,模型组大鼠关节滑膜组织中NLRP3,ASC,Caspase~(-1),L~(-1)β,IL-6,TNF-α,NF-κB表达明显升高(P<0.05),Caspase~(-1)2表达明显降低(P<0.05);与模型组比较,GT高、中剂量组NLRP3,ASC,GT各剂量组Caspase~(-1)表达水平均显著降低(P<0.05),Caspase~(-1)2表达明显升高(P<0.05),GT各组IL~(-1)β,IL-6,TNF-α,NF-κB表达均明显降低(P<0.05)。结论:GT治疗GA的作用机制可能与降低NLRP3,ASC,Caspase~(-1)表达,抑制IL~(-1)β分化成熟及NF-κB活化,降低NLRP3炎性体信号通路炎性因子表达有关。
Objective: To investigate the effect of Guizhishaoyaozhimu Tang (GT) on inflammatory signal expression in synovial tissue of rats with urate induced gouty arthritis (GA). Methods: One hundred and eighty male Sprague-Dawley rats were randomly assigned to three experiments: synovial tissue immunohistochemistry, enzyme-linked immunosorbent assay (ELISA) and Western blot. Sixty rats in each experiment were randomly divided into 6 groups according to body weight, with 10 rats in each group. They were model group, normal group, GT high, middle and low dose groups (4, 8, 16 g · kg ~ (-1) )), Colchicine positive group (3 × 10-4g · kg -1). Experimental group were ig administration, normal group, model group given equal volume of distilled water, 1 day, continuous administration of 7 d. On the fifth day before ig, rats were injected with sodium urate suspension into the foot and ankle to induce GA. The synovial tissues of rats were taken out and the expression of Nod-like receptor 3 (NLRP3) inflammasome was detected by immunohistochemistry. The mean integral absorbance (IA) was measured by Image-Pro Plus 6.0 image analysis system and the apoptosis was detected by Western blot (ASC) and Caspase ~ (-1) signal transduction proteins were detected by enzyme-linked immunosorbent assay (ELISA). The levels of interleukin- Β1, IL-6, TNF-α and NF-κB were detected by ELISA. Results: Compared with the normal group, the expression of NLRP3, ASC, Caspase -1, L -1, IL-6, TNF-α, NF (P <0.05), and the expression of Caspase 1 (-1) was significantly lower in model group than in model group (P <0.05) (P <0.05). The expression of Caspase 1 (-1) in GT group was significantly higher than that in GT group (P <0.05) NF-κB expression were significantly lower (P <0.05). CONCLUSION: The mechanism of action of GT in treating GA may decrease the expression of NLRP3, ASC and Caspase -1, inhibit the differentiation and maturation of IL-1 and the activation of NF-κB, and decrease the expression of inflammatory factors in NLRP3 inflammasome signaling pathway related.