摘要
目的:研究清热通络方通过P2X7R/NALP3/IL-1β炎症信号通路干预痛风性关节炎(GA)大鼠的作用机制。方法:60只雄性SD大鼠随机分为空白对照组,模型组,秋水仙碱组,清热通络方低、中、高剂量组(1.63、8.14、16.3 g·kg^(-1)·d~(-1))。除空白对照组、模型组外,各组大鼠预灌胃给药4 d。除空白对照组外,第4天灌胃后复制GA模型,造模后各时间段(4、6、12、24、48、72 h)检测大鼠踝关节肿胀率和热痛阈。给药7 d后取材,HE染色观察踝关节滑膜病理情况,生化法检测血清ATP水平,ELISA法测血清白细胞介素(IL)-8、IL-37、IL-1β水平,免疫组化法测滑膜组织P2X7R、NF-κB(P50、P65)、NALP3、IL-1β蛋白表达,RT-qPCR检测PBMCs P2X7R、IL-1β mRNA表达,Western Blot检测PBMCs P2X7R、IL-1β蛋白表达。结果:造模后4 h,各组大鼠踝关节肿胀率升高,24 h达高峰后下降;热痛阈下降,24 h达最低后上升。造模72 h后,与模型组比较,秋水仙碱组和清热通络方高剂量组肿胀率和热痛阈显著改善(P<0.05)。秋水仙碱及清热通络方各剂量组踝关节滑膜细胞炎症均有所改善;秋水仙碱组和清热通络方高剂量组血清ATP、IL-8、IL-37、IL-1β水平,PBMCs中P2X7R、IL-1β蛋白及mRNA表达,滑膜组织P2X7R、NALP3、NF-κB(P50、P65)、IL-1β蛋白表达均显著降低(P<0.05)。结论:清热通络方可能通过抑制GA大鼠P2X7R/NALP3/IL-1β信号通路缓解炎症,发挥治疗作用。
Objective:To investigate the role of Qingre Tongluo Formula in the treatment of rats with gouty arthritis(GA)through influencing P2X7R/NALP3/IL-1βinflammation signal pathway.Methods:Sixty male SD rats were randomly divided into a blank control group,a model group,a colchicine group,and a low-,medium-and high-dose group(1.63,8.14,16.3 g·kg^(-1)·d^(-1))of Qingre Tongluo Formula.Except for the blank and model groups,the rats in all groups were administered by pre-gavage for 4 d.The GA model was replicated on day 4 after gavage in all groups except the blank group,and the swelling rate and thermal pain domain of the ankle joint were examined at different time points(4,6,12,24,48,72 h)after modeling.The rats were continued to be administered until day 7,and HE staining was performed to observe the ankle synovial pathology.Serum ATP levels were measured by biochemical method;levels of serum IL-8,IL-37 and IL-1βwere measured by ELISA;the expression of P2X7R,NF-κB(P50,P65),NALP3 and IL-1βprotein in synovial tissues was detected by immunohistochemistry;RT-qPCR was performed to detect P2X7R,IL-1βmRNA expression in PBMCs;the expression of P2X7R,IL-1βprotein in PBMCs was measured by Western Blot.Results:The swelling rate of the ankle joint in each modeling group increased at 4 h,peaked at 24 h,and then began to decrease;the thermal pain domain decreased at 4 h,peaked at 24 h,and then tended to increase.After 72 h of modeling,the swelling rate and thermal pain domain of the rats in the colchicine group and the high-dose group of Qingre Tongluo Formula were significantly improved compared with those in the model group(P<0.05).The inflammation of ankle synovial cells in the colchicine group and each dose groups of Qingre Tongluo Formula improved;the levels of serum ATP,IL-8,IL-37,IL-1βlevels,P2X7R and IL-1βprotein and mRNA in PBMCs,the positive expression rates of P2X7R,NALP3,NF-κB(P50,P65),IL-1βprotein in synovial tissue were significantly decreased in the colchicine group and the high-dose group of Qingre Tongluo Formula(P<0.05).Conclusion:Qingre Tongluo Formula may exert therapeutic effects by alleviating inflammation through inhibiting the P2X7R/NALP3/IL-1βsignal pathway in rats with GA model.
作者
沈嘉艳
杨琦
张晓宇
谢招虎
李兆福
SHEN Jia-yan;YANG Qi;ZHANG Xiao-yu;XIE Zhao-hu;LI Zhao-fu(Yunnan University of Chinese Medicine,Kunming 650500,China;Dechang Hospital of Traditional Chinese Medicine,Liangshan 615500,China)
出处
《中华中医药杂志》
CAS
CSCD
北大核心
2023年第5期2061-2066,共6页
China Journal of Traditional Chinese Medicine and Pharmacy
基金
国家自然科学基金项目(No.81760868,No.82260925)
国家中医临床研究基地,云南省中医(风湿病)临床医学研究中心(No.202102AA310006)
云南省高层次科技人才及创新团队选拔专项-团队专项(No.202305AS350007)
云南省高校中医风湿免疫病重点实验室(No.2018YGZ01)
云南省科技计划重点项目(No.2016FA037)。
