补肾利湿法中药复方含药血清对痛风性关节炎大鼠成纤维样滑膜细胞NALP6信号转导通路调控作用机制的研究  

Regulatory effect of tonifying kidney and disinhibiting dampness therapy on fibroblast-like synoviocytes of gouty arthritis rats through NALP6 signal transduction pathway

作　　者：隋方宇[1] 韩洁茹[1] 解颖[1] 张淼[1] 姜徳友 SUI Fangyu;HAN Jieru;XIE Ying;ZHANG Miao;JIANG Deyou(Basic Medicine College,Heilongjiang University of Chinese Medicine,Harbin,Heilongjiang 150040,China)

机构地区：[1]黑龙江中医药大学基础医学院,黑龙江哈尔滨150040

出　　处：《上海中医药杂志》2022年第9期64-69,共6页Shanghai Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金项目(81173170);黑龙江省教育科学“十三五”规划重点课题(GJB1320343);黑龙江中医药大学科研基金面上项目(2019MS32)

摘　　要：目的从细胞水平探讨补肾利湿法中药复方含药血清对痛风性关节炎(GA)大鼠成纤维样滑膜细胞中嗜中性白细胞碱性磷酸酶6(NALP6)信号转导通路的调控作用。方法在大鼠成纤维样滑膜细胞培养基中加入尿酸钠微晶体刺激建立痛风性关节炎模型,将筛选出的最适合大鼠滑膜细胞生存的含药血清加入培养基中进行细胞实验。大鼠成纤维样滑膜细胞共分为空白组(正常滑膜细胞)、模型组(滑膜细胞+尿酸钠)和治疗组(滑膜细胞+尿酸钠+含药血清)3组,各组进行相应的干预。采用RT-PCR法检测滑膜细胞中NALP6 mRNA表达,Western blot法检测滑膜细胞中NALP6蛋白、核因子κB(NF-κB)蛋白、核因子κB抑制蛋白(IκB)表达,比色法检测滑膜细胞中半胱氨酸天冬氨酸蛋白水解酶-1(Caspase-1)活性,ELISA法检测各组滑膜细胞中白介素-1β(IL-1β)含量的变化。结果与空白组比较,模型组中NALP6 mRNA的表达上调,NALP6、NF-κB蛋白表达上调,IκB蛋白表达下调,Caspase-1活性提高,IL-1β含量升高,差异均有统计学意义(P<0.05)。与模型组比较,治疗组大鼠成纤维样滑膜细胞中NALP6蛋白和mRNA表达显著抑制,NF-κB蛋白表达抑制,Caspase-1的活性抑制,IL-1β的含量降低,IκB蛋白表达升高,差异均有统计学意义(P<0.05)。结论补肾利湿法中药复方含药血清抑制“NALP6-Caspase-1-IL-1β”这一炎症信号转导通路,可能是其防治痛风性关节炎的重要机制之一。Objective To explore the regulatory effect of serum containing traditional Chinese medicine compound with the function of tonifying thekidney and disinhibiting dampness on neutrophilic alkaline phosphatase 6(NALP6)signal transduction pathway in fibroblast-like synoviocytes ofgouty arthritis(GA)rats.Methods The gouty arthritis model was established by adding sodium urate microcrystalline into the rat fibroblast synovialcell culture medium.The drug containing serum which is most suitable for the survival of rat synovial cells was added into the culture medium for cellexperiment.The cells were divided into the blank group(normal synoviocytes),the model group(synoviocytes+sodium urate)and the treatmentgroup(synoviocytes+sodium urate+drug-containing serum)and each group carried on the corresponding intervention.The expression of NALP6 mRNA in synovial cells was detected by RT-PCR,the expression of NALP6,NF-κB,IκB by Western blot,the Caspase-1 activity by ColorimetricMethod and IL-1βcontent in synovial cells of each group by ELISA.Results In the model group,compared with the blank group,the expressions ofNALP6 mRNA,NALP6 and NF-κB proteins were up-regulated,while the expression of IκB protein was down-regulated.Caspase-1 activity and IL-1βcontent were increased(all P<0.05).In the treatment group,compared with the model group,the serum containing TCMcompound could significantly inhibit the expressions of NALP6 mRNA,NALP6 and NF-κB proteins,inhibit the activity of Caspase-1,reduce the content of IL-1β,and stimulate the expression of IκBprotein in fibroblast synovial cells(all P<0.05).Conclusion Blocking the“NALP6-Caspase-1-IL-1β”inflammatory signal pathwaymay be one of the most important mechanisms for preventing and treating the gouty arthritis through tonifying the kidney and disinhibiting dampness therapy.

关 键 词：痛风性关节炎 补肾利湿法 嗜中性白细胞碱性磷酸酶6 成纤维样滑膜细胞 中药研究 

分 类 号：R285.5[医药卫生—中药学]