阿托伐他汀对强直性脊柱炎患者外周血单个核细胞体外干预后Toll样受体2、Toll样受体4的表达变化研究

目的：观察天然免疫分子Toll样受体2（TLR-2）、Toll样受体4（TLR-4）在强直性脊柱炎（ankylosing spondyli-tis,AS）患者外周血单个核细胞（peripheral blood mononuclear cells,PBMC）的mRNA水平表达,研究其与AS发病的相关性,并探讨阿托伐他汀是否对AS患者TLR-2、TLR-4的表达有影响。方法：研究对象包括AS活动期患者30例和健康志愿者30例,分别抽取外周静脉血分离单个核细胞,提取细胞mRNA。采用半定量的RT-PCR方法,检测AS患者和健康志愿者外周血单个核细胞中TLR-2、TLR-4的mRNA,通过灰度分析与β-actin相比得到目的基因相对表达水平。分离12例AS活动期患者静脉血外周血单个核细胞体外培养,并给予阿托伐他汀（终浓度为0.01 mmol/L）处理,以自身非处理细胞为对照,24 h后收获细胞,提取mRNA,采用RT-PCR方法,观察阿托伐他汀对细胞TLR-2、TLR-4的mRNA表达的影响。所有数据采用配对t检验。结果：AS患者TLR-2 mRNA的相对表达水平为（1.59±1.43）,较健康志愿者（0.06±0.039）高（P〈0.05）;AS患者TLR-4 mRNA的相对表达水平为（0.60±0.73）,较健康志愿者（0.06±0.039）高（P〈0.05）;阿托伐他汀干预组TLR-2和TLR-4的mRNA表达均低于不干预对照组（P〈0.05）。结论：AS患者PBMC的TLR-2、TLR-4的表达均较健康志愿者高,提示TLR-2和TLR-4可能参与了AS的发病。阿托伐他汀可下调AS患者PBMC的TLR-2和TLR-4的表达,可能是AS新的有价值的治疗靶点之一。

Expression of Toll-like receptor 2 and Toll-like receptor 4 after intervention by Atorvastatin work on peripheral blood mononuclear cells of ankylosing spondylitis patients

Objective: To study the expression and regulation on TLR-2 and TLR-4 after intervention by Atorvastatin working on the peripheral blood mononuclear cells(PBMC) of ankylosing spondylitis patients.Methods: The study was designed to enroll 30 cases of AS and 30 cases of healthy controls.The expression levels of TLR-2 and TLR-4 in PBMC were determined by RT-PCR technical through comparing with β-actin,used t-test to compare the means.In vitro effect of Atorvastatin(0.01 mmol/L) working on PBMC of ankylosing spondylitis patients,12 cases were entrolled in.PBMC from one patient were divided into two groups.One was treated with Atorvastatin(0.01 mmol/L),another was control.The different expression levels of TLR-2 and TLR-4 with or without treated by Atorvastatin were also determined by RT-PCR method.Paired-t test was used to calculate the results.Results: The TLR-2 mRNA expression level of AS patients was higher(1.59±1.43) than control(0.06±0.039)(P<0.05).The TLR-4 mRNA expression level of AS patients was higher(0.60±0.73) than control(0.06±0.039)(P<0.05).Atorvastatin down regulated TLR-2 mRNA and TLR-4 mRNA in PBMC of AS patients in vitro test.After analysed by paired-t test,the result showed that it was more remarkable that Atorvastatin reduced the expression level of TLR-2 mRNA and TLR-4 mRNA in AS patients than control(P<0.05) respectively.Conclusion: The mRNA expression levels of TLR-2 and TLR-4 in AS patients are higher than that in controls,which may play an important role in immune reaction of AS.Atorvastatin has an effect to reduce the expression of TLR-2 mRNA and TLR-4 mRNA in PBMC of AS patients.It is possible that Atorvastatin is a potential and useful medicine of AS in future.