辣椒素对CIA大鼠关节滑膜组织抗炎作用及机制  

Study on the anti-inflammatory mechanism of capsaicin on CIA rats synovia

作　　者：李雨 王杰 陈铭勰 艾丽霞 张耀文 何子晗 吴遵秋 李蓉 LI Yu;WANG Jie;CHEN Mingxie;AI Lixia;ZHANG Yaowen;HE Zihan;WU Zunqiu;LI Rong(School of Basic Medical Science,Guizhou Medical University,Guiyang 550025,Guizhou,China)

机构地区：[1]贵州医科大学基础医学院,贵州贵阳550025

出　　处：《贵州医科大学学报》2023年第5期508-514,共7页Journal of Guizhou Medical University

基　　金：国家级大学生创新创业项目(202010660018);贵州省大学生创新创业项目(202110660037);国家自然科学基金项目(82060776);贵州省科技计划项目(黔科合基础[2020]1Y388)。

摘　　要：目的探究辣椒素(CAP)对胶原诱导性关节炎(CIA)大鼠关节滑膜组织抗炎作用机制。方法48只SD大鼠均分为空白组(Nor组)、模型组(Mod组)、雷公藤组(GTW组)及CAP高(3 mg/kg)、中(2 mg/kg)、低剂量(1 mg/kg)组,后5组大鼠采用牛Ⅱ型胶原和不完全弗氏佐剂(IFA)建立CIA大鼠模型,Nor组及Mod组大鼠灌胃生理盐水、另4组大鼠灌胃相应药物3周,测定大鼠足肿胀度及关节炎指数(AI)评分;取大鼠右踝滑膜组织HE染色观察组织学变化,采用酶联免疫吸附(ELISA)法检测大鼠血清中IL-17含量,实时荧光定量PCR(RT-qPCR)技术检测大鼠滑膜组织中丝裂原活化蛋白激酶激酶3(MKK3)、P38丝裂原活化蛋白激酶(P38MAPK)、基质金属蛋白酶13(MMP-13)、半胱氨酸蛋白酶-3(Caspase-3)mRNA表达,蛋白免疫印迹(Western blot)法检测磷酸化丝裂原活化蛋白激酶激酶3(p-MKK3)、磷酸化P38丝裂原活化蛋白激酶(p-P38MAPK)、MMP-13和Caspase-3蛋白表达水平。结果与Nor组比较,Mod组大鼠足肿胀度以及AI评分升高(P<0.001);与Mod组比较,GTW组和CAP各剂量组大鼠足肿度及AI评分降低(P<0.01);GTW组和CAP高、低剂量组大鼠踝关节组织炎性细胞浸润、成纤维滑膜细胞增生均减少;GTW组和CAP高剂量组血清IL-17降低(P<0.01),CAP中剂量组血清IL-17降低(P<0.05);GTW组和CAP高剂量组大鼠滑膜关节组织中MKK3、P38MAPK和MMP-13 mRNA降低(P<0.05),Caspase-3 mRNA升高(P<0.01);GTW组和CAP高剂量组滑膜关节组织中p-MKK3、p-P38MAPK和MMP-13蛋白表达降低(P<0.01),Caspase-3蛋白表达升高(P<0.01)。结论CAP对CIA模型大鼠关节滑膜组织的抗炎作用机制可能与抑制P38MAPK信号通路活化、下调促炎因子IL-17和上调促凋亡因子Caspase-3的表达有关。Objective To investigate the anti-inflammatory mechanism of capsaicin(CAP)on collagen induced arthritis(CIA)rats synovia.Methods A total of 48 SD rats were evenly divided into blank group(Nor group),model group(Mod group),tripterygium wilfordii group(GTW group),and CAP high dose group(3 mg/kg),medium dose group(2 mg/kg),and low dose group(1 mg/kg).The last 5 groups of rats were used to establish a CIA rat model using bovine type II collagen and incomplete Freund's adjuvant(IFA).The Nor group and Mod groups were given intragastric administration of normal saline,while the other 4 groups were given drugs for 3 weeks,detecting the degree of foot swelling and arthritis index(AI)score of all groups.HE staining was performed to detect morphological changes of rat's right ankle joint.In addition,serum IL-17 content was detected by enzyme linked immunosorbent assay(ELISA).Real time fluorescent quantitative PCR(RT-qPCR)was adopted to detect mRNA expression of MKK3,P38MAPK,MMP-13,Caspase-3 in rat's synovia;Western blot was used to detect protein expression levels of p-MKK3,p-P38MAPK,MMP-13,and Caspase-3.Results Compared with Nor group,the swelling degree of rat's foot and AI index score of Mod group were increased(P<0.001).Compared with the Mod group,the swelling degree and AI score were decreased in GTW group and CAP groups of varied dosages(P<0.01).Inflammatory cell infiltration and fibroblast synovial cell proliferation were decreased in the GTW group,CAP highdose group,and CAP lowdose group.The levels of serum IL-17 in the GTW group and CAP highdose group were decreased(P<0.01),and that in the CAP mediumdose group was decreased(P<0.05).The expressions of MKK3,P38MAPK,and MMP-13 mRNA in rat's synovia were decreased in the GTW group and CAP highdose group(P<0.05),while the expressions of Caspase-3 mRNA were increased(P<0.01).The protein expressions of p-MKK3,p-P38MAPK,and MMP-13 were decreased in the GTW group and CAP highdose group(P<0.01),while the protein expressions of Caspase-3 were increased(P<0.01).Conclusion The

关 键 词：类风湿性关节炎 辣椒素 P38MAPK信号通路 IL-17 半胱氨酸蛋白酶-3 大鼠 

分 类 号：R34[医药卫生—基础医学]