分析处于不同进展阶段的狼疮肾炎(LN)患者循环miRNA的表达变化并探讨其可能存在的临床意义。

本研究共94份血清标本，包括58例LN患者和36名健康体检者(作为对照组)。挑选出其中性别、年龄匹配的12份血清标本用于循环miRNA表达谱筛选，包括早期LN组4份(CKD 1～3期)，晚期LN组4份(CKD分期4～5期)及对照组4份作为芯片组，使用miRNA PCR芯片检测血清miRNA变化；剩余50例LN患者(40例早期LN患者、10例晚期LN患者)及32名对照者血清样本作为验证组，用于验证miRNA表达谱筛选结果；以Nanodrop 2000检测血清总RNA抽提浓度；选择miR-130b-3p和miR-1233-3p作为验证对象；进行Spearman相关性分析。

芯片组结果显示，与对照组相比，早期LN组中7个表达上升的miRNA差异具有统计学意义，分别是：miR-1233-3p (P＝0.019)、miR-130b-3p (P＝0.021)、miR-18a-3p (P＝0.021)、miR-628-3p (P＝0.023)、miR-1260b (P＝0.030)、miR-1539 (P＝0.041)和miR-378e (P＝0.047)；晚期LN组分别与对照组及早期LN组比较，未发现表达上调大于2倍或者上调差异具有统计学意义的miRNA。验证组结果显示：晚期LN组患者分别与对照组、早期LN组比较，血清总RNA浓度下降(U＝4.5，P<0.001；U＝18.0，P<0.001)；早期LN组与对照组比较，血清中miR-130b-3p表达升高[IQR 16.2(8.7，42.7)与9.6(4.8，17.4), U＝405.5，P＝0.008]；晚期LN组分别与对照组及早期LN组比较，MiR-130b-3p和miR-1233-3p表达均下降(U＝69.0、P＝0.008，U＝46.0、P<0.001；U＝80.0、P＝0.019，U＝70.0、P＝0.002)；相关性分析显示循环miR-130b-3p的相对表达量与24 h尿蛋白(r＝0.404，P＝0.010)、肾脏慢性活动指数(r＝0.389，P＝0.013)、甘油三酯(r＝0.376，P＝0.017)呈正相关。

严重肾功能衰竭患者可能存在循环miRNA广泛表达下调，循环miRNA-130b-3p在早期LN患者中表达升高并与肾脏损伤和血脂调节异常相关，在LN发生发展中可能起作用。

To analyze the changes of circulating miRNAs expression and explore their possible significance in patients with lupus nephritis (LN) at different stages.

There were a total of 94 serum samples from 58 LN patients and 36 healthy controls. 12 serum samples selected that were comparable in gender and age were used to analyze the expression profile of circulating miRNAs with PCR array, including serum samples (n=4 in each group) from healthy control, early stage LN patients (CKD 1-3 stage), and late stage LN patients (CKD 4-5 stage); and the other 50 LN patients were used as the validation group, including 40 cases of early LN patients, and 10 late LN patients. The Nanodrop 2000 was were determined by the Spearman rank correlation coefficient.

The used for detecting RNA concentration. Two miRNAs, miR-130b-3p and miR-1233-3p were selected as to be validated. Correlations expressions of seven miRNAs, including miR-1233-3p (P=0.019), miR-130b-3p (P=0.021), miR-18a-3p (P=0.021), miR-628-3p (P=0.023), miR-1260b (P=0.030), miR-1539 (P=0.041), and miR-378e (P=0.047) were increased in early stage LN patients compared with the healthy controls. On the other hand, the expression levels of most circulating miRNAs were found decreased in late stage LN patients, and no circulating miRNAs were found up-regulated more than 2 times or increased significantly compared to those in the healthy controls or early LN patients; the tatol serum RNA was reduced in late LN patients of validation group compared to that in healthy controls (P<0.001, U=4.5) or early LN patients (P<0.001, U=18.0); the level of miR-130b-3p was significantly higher in early stage LN patients compared to healthy controls [IQR 16.2 (8.7, 42.7) vs 9.6 (4.8, 17.4), P=0.008, U=405.5)], and decreased significantly in late stage LN group compared to healthy controls (U=69.0, P=0.008) or early LN patients (U=46.0, P<0.001). The miR-1233-3p also significantly down-regulated in late stage LN group compared to healthy controls (U=80.0, P=0.019) or early LN patients (U=70.0, P=0.002). The expression of serum miR-130b-3p was significantly and positively correlated with 24-hour proteinuria (r=0.404, P=0.010), chronicity index (r=0.389, P=0.013), and serum triglyceride level (r=0.376, P=0.017).

The circulating miRNA levels were reduced in patients with severe chronic renal failure. MiR-130b-3p was up-regulated in early stage LN patients, and associated with renal injury and abnormal regulation of serum lipids, playing a possible role in the development and progression of LN.