基于NLRP3炎性小体通路探讨电针对急性痛风性关节炎大鼠组织蛋白酶B的影响  

Cathepsin-B involved in effect of electroacupuncture by inhibiting the activation of NLRP3 inflammasome in rats with acute gouty arthritis

作　　者：谯童茜 唐成林[1] 邱丽 赵丹丹[1] 吴梦佳[1] 安荟羽 马翔 万小凤 QIAO Tong-xi;TANG Cheng-lin;QIU Li;ZHAO Dan-dan;WU Meng-jia;AN Hui-yu;MA Xiang;WAN Xiao-feng(College of Traditional Chinese Medicine,Chongqing Medical University,Chongqing 400016,China)

机构地区：[1]重庆医科大学中医药学院,重庆400016

出　　处：《针刺研究》2021年第4期295-300,共6页Acupuncture Research

基　　金：国家自然科学基金项目(No.81273870)。

摘　　要：目的:观察电针对大鼠膝关节滑膜中组织蛋白酶B(Cathepsin-B)的影响,探讨电针治疗急性痛风性关节炎的机制。方法:60只Wistar大鼠随机分为空白组、模型组、药物组和电针组,每组15只。采用左膝关节腔注射0.2 mL尿酸钠晶体混悬液法建立急性痛风性关节炎大鼠模型。药物组予秋水仙碱灌胃治疗(0.3 mg·kg^(-1)·d^(-1)),电针组予左侧"三阴交""足三里"电针治疗,10 min/次,1次/d,共1周。用Coderre步态分级标准对各组大鼠进行步态评分;HE染色观察大鼠左膝关节滑膜组织的形态学变化;Western blot法检测大鼠左膝关节滑膜Cathepsin-B的表达;实时荧光定量PCR法检测大鼠左膝关节滑膜组织中NOD样受体蛋白3(NLRP3)、凋亡相关斑点样蛋白(ASC)和半胱氨酸天冬氨酸蛋白酶1(Caspase-1)、白细胞介素-1β(IL-1β)、IL-18 mRNA的表达。结果:与空白组比较,模型组大鼠左膝关节滑膜有明显炎性浸润,步态评分以及Cathepsin-B蛋白和NLRP3、ASC、Caspase-1、IL-1β、IL-18 mRNA表达水平显著增高(P<0.01)。与模型组比较,药物组炎性浸润程度较轻,电针组稍次之;药物组和电针组步态评分以及Cathepsin-B蛋白和NLRP3、 ASC、Caspase-1、IL-1β、IL-18 mRNA表达水平均显著降低(P<0.01,P<0.05)。与药物组比较,电针组Caspase-1、IL-18 mRNA表达水平增高(P<0.05)。结论:电针可能是通过降低膝关节滑膜中Cathepsin-B的活性来抑制NLRP3炎性小体的激活,从而治疗急性痛风性关节炎。Objective To observe the effect of electroacupuncture(EA) on Cathepsin-B in the synovium of the knee joint of acute gouty arthritis(AGA) rats, so as to explore the mechanism of EA in the treatment of AGA. Methods A total of 60 male Wistar rats were randomly divided into normal control, model, medication and EA groups, with 15 rats in each group. Rat model of AGA was established by injection of 0.2 mL sodium urate crystal suspension into the left knee joint cavity. The rats in the medication group were treated with colchicine by gavage(0.3 mg·kg^(-1)·d^(-1)), and the rats in the EA group were treated with EA at the left "Sanyinjiao"(SP6) and "Zusanli"(ST36) for 10 min each time, once a day for a week. The Coderre gait grading standard was used to score the gait of rats. The pathological morphology of synovial tissue of the left knee joint was observed by H.E. staining. The expression levels of Cathepsin-B protein and Nod-like receptor pyrin domain 3(NLRP3), apoptosis-associated speck-like protein containing CARD(ASC),Caspase-1, interleukin-1β(IL-1β) and IL-18 mRNAs were detected by Western blot and real-time fluorescence quantitative PCR, respectively. Results Compared with the normal control group, the degree of synovitis infiltration in the model group was more serious. And the gait score, the protein expression level of Cathepsin-B and the mRNA expression levels of NLRP3,ASC,Caspase-1, IL-1β,IL-18 were significantly increased(P<0.01).After the interventions, the degree of inflammatory infiltration was mild, The gait score, the protein expression level of Cathepsin-B and the mRNA expression levels of NLRP3 and ASC,Caspase-1,IL-1β,IL-18 were significantly decreased in both medication and EA groups in contrast to the model group(P<0.01, P<0.05). Compared with medication group, the mRNA expression levels of Caspase-1 and IL-18 in the EA group were increased(P<0.05). Conclusion EA may inhibit the activation of NLRP3 inflammasome by reducing the activity of Cathepsin-B in the synovium of the knee joint, so as t

关 键 词：急性痛风性关节炎 电针 NOD样受体蛋白3 组织蛋白酶B 

分 类 号：R245.97[医药卫生—针灸推拿学]