复方土茯苓颗粒对大鼠滑膜细胞炎症因子及mi-RNA的影响  

Effect of Compound Rhizoma Smilacis Glabrae Granules on Inflammatory Factors and mi-RNA in Synovial Cells of Rats

作　　者：朱明敏[1,2] 李静[1,2] 张欢欢[2] 刘颖琬[1] 孙维峰[1] 

机构地区：[1]广州军区广州总医院,广东广州510010 [2]广州中医药大学,广东广州510006

出　　处：《广州中医药大学学报》2014年第4期578-581,586,678,共6页Journal of Guangzhou University of Traditional Chinese Medicine

基　　金：国家自然科学基金资助项目(编号:81072915);广东省自然科学基金资助项目(编号:s2012010009032);广东省科技计划重大专项(编号:2012A080201012)

摘　　要：【目的】观察复方土茯苓颗粒含药血清对大鼠滑膜细胞痛风模型的影响以及与白细胞介素-1β/白细胞介素-6(IL-1β/IL-6)、小片段RNA miR-146a的关系。【方法】从SD大鼠身上摘取膝关节滑膜组织进行原代培养并传代,将滑膜细胞分成空白对照组,空白血清组,模型组,中药血清低、中、高剂量组6组;分别制备复方土茯苓颗粒中药血清和尿酸盐晶体混悬液,并进行细胞活性试验后,按组进行相应的干预,然后收集细胞培养液采用酶联免疫吸附反应(ELISA)法检测IL-1β/IL-6水平;提取滑膜细胞总RNA并进行逆转录—聚合酶链反应(RT-PCR)检测miR-146a表达,并分析它们的关系。【结果】模型组滑膜细胞培养液中IL-1β/IL-6的含量显著升高(P<0.05),滑膜细胞miR-146a表达显著下调(P<0.05);中药血清各剂量组IL-1β/IL-6的含量显著降低(P<0.05),miR-146a则显著上调(P<0.05);IL-1β/IL-6与miR-146a表达呈反比关系。【结论】复方土茯苓颗粒含药血清可能是通过上调大鼠滑膜细胞miR-146a的表达,抑制IL-1β/IL-6,从而缓解炎症反应。Objective To study the effect of serum containing Compound Rhizoma Smilacis Glabrae Granules （CRSGG） on interleukin 1β （IL-1β） , IL-6 and miR-146a in synovial cells of rats, and to explore the relationship among IL-1β, IL-6 and microRNA of miR-146a. Methods The knee synovial tissues were isolated from SD rats for primary culture and subculture. And then the cultured synovial cells were divided into blank control group, blank serum control group, model serum control group, and low-, middle- and high-dose CRSGG serum groups. Serum containing CRSGG and urate crystal suspension were prepared for the coeulture with the synovial cells. After culture with the corresponding serum or suspension, IL-1β and IL-6 levels in the supernatant were detected by enzyme-linked immunospecifie assay （ELISA） , and miR-146a expression in the total RNA of synovial cells was detected by reverse transcription polymerase chain reaction （RT-PCR） . Results In model serum control group, contents of IL-1β and IL-6 were significantly higher, and expression level of miR-146a was significantly lower than the blank serum control group （P〈0.05） . In CRSGG serum groups, contents of IL-1β and IL-6 were significantly lower, and expression level of miR-146a was significantly higher than the model serum control group （P〈0.05） . An inverse relationship was presented between IL-1β/IL-6 and miR-146a. Conclusion CRSGG has an effect on relieving inflammatory reaction through up-regulating miR- 146a expression of synovial cells and by decreasing IL-1β/IL-6.

关 键 词：复方土茯苓颗粒 药理学 痛风 中药疗法 细胞培养 基因表达调控 滑膜细胞 病理学 疾病模型 动物 大鼠 

分 类 号：R285.5[医药卫生—中药学]