摘要
目的探讨丁香脂素(Syr)在Ⅱ型胶原诱导关节炎(CIA)大鼠中的作用及相关机制。方法从70只雄性Wistar大鼠中随机选取12只不做任何处理作为Normal组,另58只大鼠均经皮下注射Ⅱ型胶原制作CIA模型,将其中造模成功且关节炎指数(AI)接近的48只大鼠随机分为Control组、Syr20组、Syr50组和Syr100组,每组12只。造模成功后,Syr20组、Syr50组和Syr100组大鼠分别接受Syr 20、50、100 mg·kg^(-1)·d^(-1)连续灌胃36 d,Normal组、Control组大鼠接受相同体积的生理盐水连续灌胃36 d。在Syr或生理盐水给药前1 d和给药后第6、12、18、24、30、36 d对5组大鼠分别进行AI评定。采用酶联免疫吸附试验(ELISA)检测5组大鼠血清谷胱甘肽(GSH)、过氧化氢酶(CAT)、超氧化物歧化酶(SOD)、白细胞介素(IL)-6、IL-1β、肿瘤坏死因子(TNF)-α水平。采用逆转录聚合酶链反应(RT-PCR)检测5组大鼠关节组织促炎因子IL-6、IL-1β、TNF-αmRNA表达水平。采用蛋白质免疫印迹法(Western blot)检测5组大鼠关节组织Toll样受体4(TLR4)、核转录因子(NF)-κB p65、基质金属蛋白酶(MMP)-3水平。结果Control组大鼠生理盐水给药后第6、12、18、24、30、36 d的AI均高于Normal组(P<0.01)。Syr20组、Syr50组及Syr100组大鼠Syr给药后第6、12、18、24、30、36 d的AI均低于Control组(P<0.05或P<0.01)。Syr50组大鼠Syr给药后第12、30 d及Syr100组大鼠Syr给药后第6、12、30 d的AI均低于Syr20组(P<0.05或P<0.01)。Control组大鼠血清GSH、CAT、SOD水平均低于Normal组,IL-6、IL-1β、TNF-α水平均高于Normal组(P<0.01)。Syr20组、Syr50组及Syr100组大鼠血清GSH、CAT、SOD水平均高于Control组,Syr50组及Syr100组大鼠血清IL-6、IL-1β、TNF-α水平均低于Control组(P<0.05或P<0.01)。Control组大鼠关节组织IL-6、IL-1β、TNF-αmRNA表达水平均高于Normal组(P<0.01)。Syr20组大鼠关节组织TNF-αmRNA及Syr50组、Syr100组大鼠关节组织IL-6、IL-1β、TNF-αmRNA表达水平均低于Control组(P<0.05或P<0.01)。Control组大鼠关节组织TLR4、NF-κB p65、MMP-3水平均高于Normal组,Syr20组、Syr50组、Syr100组大鼠上述指标均低于Control组(P<0.05或P<0.01)。结论Syr可能通过抑制TLR4/NF-κB信号通路缓解CIA大鼠的关节炎症状。
Objective To explore the effect of syringaresinin(Syr)on rats of typeⅡcollagen-induced arthritis(CIA)and related mechanism.Methods From 70 male Wistar rats,12 rats were randomly selected as Normal group without any treatment,and the other 58 rats were subcutaneously injected with typeⅡcollagen to make CIA model.Forty-eight rats with making model successfully and similar arthritis index(AI)were randomly divided into Control group,Syr20 group,Syr50 group and Syr100 group,with 12 rats in each group.After making model successfully,rats in Syr20 group,Syr50 group and Syr100 group received Syr 20,50 and 100 mg·kg^(-1)·d^(-1)respectively by intragastric administration for 36 d continuously.Rats in Normal group and Control group received same volume of saline by intragastric administration for 36 d continuously.AI was assessed on 1 d before administration of Syr or saline and on the 6 th,12 th,18 th,24 th,30 th and 36 th day after administrationof Syr or saline respectively.Levels of serum glutathione(GSH),catalase(CAT),superoxide dismutase(SOD),interleukin(IL)-6,IL-1βand tumor necrosis factor(TNF)-αin rats of the 5 groups were detected by enzyme-linked immunosorbent assay(ELISA).The expression levels of pro-inflammatory factors IL-6,IL-1βand TNF-αmRNA in joint tissues of rats of 5 groups were detected by reverse transcriptase polymerase chain reaction(RT-PCR).Levels of toll-like receptor 4(TLR4),nuclear transcription factor(NF)-κB p65 and matrix metalloproteinase(MMP)-3 were detected by Western blotting.Results AI of rats in Control group was higher than that in Normal group on the 6 th,12 th,18 th,24 th,30 th and 36 th day after administration of saline(P<0.01).AI of rats in Syr20 group,Syr50 group and Syr100 group on the 6 th,12 th,18 th,24 th,30 th,36 d after administration of Syr were lower than those in Control group(P<0.05 or P<0.01).AI of rats in Syr50 group on the 12 th and 30 th day after administration of Syr and in Syr100 group on the 6 th,12 th and 30 th day after administration of Syr were lower than those in Syr20 group(P<0.05 or P<0.01).Levels of serum GSH,CAT and SOD in Control group were lower than those in Normal group,levels of IL-6,IL-1βand TNF-αwere higher than those in Normal group(P<0.01).Levels of serum GSH,CAT and SOD in Syr20 group,Syr50 group and Syr100 group were higher than those in Control group,levels of serum IL-6,IL-1βand TNF-αin Syr50 group and Syr100 group were lower than those in Control group(P<0.05 or P<0.01).Expression levels of IL-6,IL-1βand TNF-αmRNA in joint tissues of rats in Control group were higher than those in Normal group(P<0.01).Expression levels of TNF-αmRNA in joint tissues of rats in Syr20 group and IL-6,IL-1β,TNF-αmRNA in joint tissues of rats in Syr50 group and Syr100 group were lower than those in Control group(P<0.05 or P<0.01).Levels of TLR4,NF-κB p65 and MMP-3 in joint tissues of rats in Control group were higher than those in Normal group,above indexes of rats in Syr20 group,Syr50 group and Syr100 group were lower than those in Control group(P<0.05 or P<0.01).Conclusion Syr may relieve the symptoms of arthritis in rats of CIA by inhibiting TLR4/NF-κB signaling.
作者
潘明明
冯燕
李青梅
Pan Mingming;Feng Yan;Li Qingmei(Department of Osteoporosis,Xi’an Red Cross Hospital,Xi’an 710068,China)
出处
《临床内科杂志》
CAS
2022年第10期696-700,共5页
Journal of Clinical Internal Medicine
基金
陕西省重点研发项目(S2018-YYF-YBSF-0219)
