健脾滋肾法联合西药治疗系统性红斑狼疮对患者血浆microRNA的影响

Effects of spleen invigorating and kidney moistening therapy combined with western medicine on plasma microRNA level in patients with systemic lupus erythematosus

目的:基于microRNA基因芯片技术,客观评价健脾滋肾法结合西药对系统性红斑狼疮患者microRNA的调控,探讨其可能的分子机制。方法:5例符合纳入标准的SLE患者作为研究对象,予四君子汤合知柏地黄丸+泼尼松片+硫酸羟氯喹治疗9个月,观察指标:SLEDAI、系统性红斑狼疮症状分级量化表、补体、ds-DNA、24h尿蛋白、血常规、肝功能、肾功能。健康志愿者5名作为对照。抽取健康对照组及治疗组治疗前后的空腹静脉血5mL,提取血浆总RNA,Agilent human microRNA微阵列芯片试验分析microRNA表达情况,对差异microRNA进行生物学分析。结果:5例患者SLEDAI评分、系统性红斑狼疮症状分级量化评分治疗后显著下降(P<0.05);治疗前与健康者相比,得到显著差异的microRNA 58个,其中上调10个,下调48个,治疗后与治疗前相比,上述58个microRNA中有1个microRNA上调,为mi R-22-3p,3个microRNA下调,分别为mi R-1246、mi R-4644、mi R-6739-5p;对差异microRNA进行生物学分析,得出与SLE密切相关的信号通路,依次是泛素蛋白酶系统、TGF-β信号通路、Notch信号通路、p53信号通路。结论:健脾滋肾法联合西药治疗SLE安全有效;其分子机制可能是通过影响相关microRNA的表达调控某些与细胞凋亡、细胞因子、物质代谢相关的信号通路,达到调节免疫、改善体质的作用。

Objective： To objectively evaluate the regulaUon oi spleen invigorating ana Klaney moistening merapy combined with western medicine on the plasma microRNA level in patients with systemic lupus erythematosus （SLE） based on microarray technology. Methods： Five SLE patients who were in accordance with the inclusion criteria were treated respectively with Sijunzi Decoction with Zhibai Dihuang Pills combined with Prednisone Tablet and Hydroxychloroquine Sulphate for 9 months. The efficacy indicators included SLEDAI, SLE syndrome grading scale, complement, ds-DNA, 24-hour urinary protein, blood routine, liver function and renal function. Five healthy volunteers were selected as controls. Five mL of fasting venous blood was collected from the healthy control group and the treatment group before and after treatment to extract the total RNA for microRNA gene chip detection; the Agilent human microRNA microarray was used to analyze the expression of miRNA, and the biological analysis of the differential microRNA was carried out. SPSS 17.0 software was used for the statistical analysis. Results： After treatment, 5 patients＇ SLEDAI scores and the SLE syndrome grading scale were significantly reduced （P〈0.05）. Compared with the healthy volunteers before treatment, 58 microRNAs with significant differences were obtained in the treatment group, including 10 up-regulated ones and 48 down-regulated ones; compared before and after treatment, one of the up-regulated miRNA in those 58 microRNAs was miR-22-3p, while 3 down-regulated miRNAs are miR-1246, miR-4644 and miR-6739-5p respectively; the target gene prediction for differential microRNAs and the enrichment analysis concluded 4 signaling pathways closely associated with SLE, namely, ubiquitin-proteasome system, TGF- a signaling pathway, Notch signaling pathway and p53 signaling pathway. Conclusion： Spleen invigorating and kidney moistening therapy combined with western medicine is safe and effective in the treatment of SLE. The molecular mechanism of the spleen invigorating and kidney moistening therapy combined with western medicine in the SLE treatment may be by regulating the expression of related microRNA to regulate some signal pathways related to cell apoptosis, cytokine and substance metabolism, so as to achieve the function of regulating immunity and improving the constitution.