摘要
目的探讨通痹七物方对痛风性关节炎(Gouty Arthritis,GA)大鼠的防治作用及对NOD样受体蛋白3(NOD-like receptor protein 3,NLRP3)炎症小体、线粒体自噬蛋白的影响。方法将36只大鼠随机分为空白对照组,模型组,秋水仙碱组,通痹七物方低、中、高剂量组,每组6只。空白对照组、模型组给予同体积生理盐水,秋水仙碱组给予秋水仙碱0.3 mg·kg^-1水溶液,通痹七物方低、中、高剂量组分别给予通痹七物方浓缩配方颗粒800、1600、3200 mg·kg^-1水溶液。上述各组连续灌胃给药7 d。第5天给药后,除空白对照组外,其余各组大鼠右膝关节腔注射0.1 mL单钠尿酸盐(Monosodium urate,MSU)结晶溶液,进行模型制备。以上各组在模型制备24 h、48 h后进行步态评分,实验结束观察病理组织形态,检测血清肿瘤坏死因子α(TNF-α)、白细胞介素6(IL-6)的含量,检测NLRP3、半胱氨酸蛋白酶1(Caspase-1)、白细胞介素1β(IL^-1β)、自噬微管相关蛋白轻链3B(Autophagy microtubule associated protein light chain 3B,LC3B)、P62、抗增殖蛋白2(PHB2)蛋白的表达。结果与空白对照组比,模型组的步态评分明显上升,滑膜病理提示细胞增生、间质高度水肿伴大量中性粒细胞浸润,血清TNF-α、IL-6水平升高。滑膜NLRP3、Caspase-1、IL^-1β、LC3B、P62、PHB2蛋白表达量不同程度增加,差异有统计学意义(P<0.01,P<0.05)。与模型组相比,秋水仙碱组,通痹七物方低、中、高剂量组上述指标均有不同程度的改善,差异有统计学意义(P<0.01,P<0.05)。结论通痹七物方能够防治痛风性关节炎,其作用机制可能与调控线粒体自噬,抑制NLRP3炎症小体激活有关。
Objective To explore the preventive and therapeutic effects of Tongbi Qiwu formula on NLRP3 inflammasome and protein of mitochondrial autophagy in gouty arthritis(GA)rats.Methods A total of 36 SD rats were randomly divided into blank control group(K group),model group(M group),colchicine group(group Q),Tongbi Qiwu formula low dose group(T1 group),and Tongbi Qiwu formula medium dose group(T2 group),Tongbi Qiwu formula high dose group(T3 group),6 rats in each group.The K group and the M group were given the same volume of saline;the Q group was given colchicine 0.3 mg·kg^-1 aqueous solution;T1 group,T2 group,and T3 group were given concentrated granules of Tongbi Qiwu formula 800 mg·kg^-1,1600 mg·kg^-1,and 3200 mg·kg^-1 aqueous solution.The above groups were administered orally for 7 days.After the 5 thday of administration,except for the K group,rats in each group were injected with 0.1 mL of Monosodium urate(MSU)solution in the right knee joint cavity for model preparation.The gait scores of rats were obtained 24 h and 48 h after model preparation.At the end of the experiment,the pathology of rats’synovial membrane was observed,serum levels of TNF-αand IL-6 were detected,and the expression levels of NLRP3,Caspase-1,IL^-1β,LC3 B,P62 and PHB2 protein were detected by Western Blot.Results Compared with the K group,the gait score of the M group was significantly increased.Synovial pathology showed cell proliferation,interstitial highly edema with a large number of neutrophils infiltration,and increased serum TNF-αand IL-6 levels,and the expression of NLRP3,Caspase-1,IL^-1β,LC3 B,P62,PHB2 proteins in the synovial membrane was increased,all with statistical significance(P<0.01,P<0.05).Compared with the M group,the above-mentioned indicators of the Q group,T1 group,T2 group,and T3 group have different degrees of improvement,and with statistical significance(P<0.01 or P<0.05).Conclusion Tongbi Qiwu formula can treat gouty arthritis,and its mechanism may be related to regulating mitochondrial autophagy and inhibiting NLRP3 inflammasome activation.
作者
范为民
FAN Weimin(Traditional Chinese Medicine Department,Yijishan Hospital Affiliated to Wannan Medical College,Wuhu 241001 Anhui,China;School of Traditional Chinese Medicine,Southern Medical University,Guangzhou 510515 Guangdong,China)
出处
《中药新药与临床药理》
CAS
CSCD
北大核心
2020年第7期769-774,共6页
Traditional Chinese Drug Research and Clinical Pharmacology
基金
中国博士后科学基金项目(2019M652984)
广东省中医药局科研项目(20201222)
弋矶山医院引进人才项目(YR201809)。
