microRNA-141与HMGB1通路在不同程度心肾综合征中的表达及临床意义

Expression and clinical significance of microRNA-141 and HMGB1 pathway in different degrees of cardiorenal syndrome

目的研究心肾综合征(cardiorenal syndrome, CRS)患者外周血中microRNA-141(miR-141)与重组人高迁移率族蛋白B1(HMGB1)通路的表达,探讨其与CRS发生、发展之间的关系。方法收集2017年1月至2018年12月入住东营市胜利油田中心医院重症医学科的CRS患者80例及同期体检中心40例健康志愿者外周血标本,按照2010年改善全球肾脏病预后组织/急性透析质量倡议(KDIGO/ADQI)共识将患者分为急性心肾综合征(CRS Ⅰ型)和慢性心肾综合征(CRS Ⅱ型)各40例。检测血清B型钠尿肽前体(NT-proBNP)、超敏C反应蛋白(hs-CRP)、血肌酐(SCr)、左室射血分数(LVEF)、二尖瓣口舒张早期峰值血流速度(E峰)、舒张晚期峰值血流速度(A峰),计算E/A比值。运用RT-PCR法及Western blot法检测外周血标本中miR-141、HMGB1、 Beclin-1及自噬标志物(LC3)蛋白的表达情况并进行对比。结果 CRS I型患者常规指标NT-proBNP、hs-CRP、SCr、LVEF、E/A比值均高于CRSⅡ型患者,差异有统计学意义(P<0.05);RT-PCR结果显示,miR-141表达量在正常组、CRSⅠ组及CRSⅡ组分别为(1.64±0.17)、(0.58±0.12)及(1.21±0.28),两个研究组表达均低于正常组,且CRS Ⅰ组表达最低(P<0.05);HMGB1-mRNA在正常组、CRSⅠ组及CRSⅡ组中表达水平分别为(0.81±0.31)、(2.23±0.35)及(1.61±0.42),两个研究组表达均高于正常组,且CRSⅠ组表达最高(P<0.05);Western blot结果显示, HMGB1、Beclin-1和LC3在CRSⅠ型中的表达量分别为(1.69±0.21)、(1.89±0.35)、(1.94±0.28),均高于CRSⅡ型(1.31±0.29)、(1.39±0.29)、(1.34±0.31),差异有统计学意义(P<0.05),且两组表达均高于正常组(0.61±0.23)、(0.64±0.12)、(0.51±0.22),差异均有统计学意义(P<0.05)。结论 miR-141在CRSⅠ型患者血清中表达下降,与miR-141通过影响HMGB1的水平从而影响自噬水平,进而导致CRS的发生、发展有关,故miR-141可能为CRS潜在的治疗靶点。

Objective To study the expression of microRNA - 141 ( miR - 141 ) and recombinant human high mobility group protein Bl ( HMGB1) pathway in peripheral blood of patients with cardiorenal syndrome ( CRS), and to explore the relationship between miR - 141 and the occurrence and development of CRS. Methods The peripheral blood samples of 80 patients with CRS admitted to ICU in our hospital from January 2017 to December 2018 and 40 healthy volunteers in the physical examination center at the same time were collected. According to the 2010 KDIGO/ADQI consensus, the patients were divided into acute cardiorenal syndrome (CRS I ) and chronic cardiorenal syndrome ( CRS II )? Serum B - type natriuretic peptide precursor ( NT - proBNP), high sensitive C 一 reactive protein (hs - CRP), serum creatinine ( SCr), left ventricular ejection fraction ( LVEF), mitral orifice diastolic peak flow velocity (E peak) and late diastolic peak flow velocity ( A peak) were measured, and the ratio of Em A was calculated. The expression of miR - 141, HMGB1, Beclin 一 1 and LC3 in peripheral blood samples were detected by RT 一 PCR and WB. Results The ratio of NT 一 proBNP, hs 一 CRP, SCr, LVEF, E/A ratio of CRS I patients were higher than those of CRS U patients and the difference was statistically significant ( P <0. 05). The results of RT 一 PCR showed that the expression of miR - 141 in normal group, CRS I group and CRS U group were (1.64 ± 0. 17 ),( 0. 58 ± 0. 12 ) and (1.21 ± 0. 28), respectively, the expression of miR - 141 in both CRS groups was lower than that in normal group, and the expression of miR - 141 in CRS I group was the lowest ( P V 0. 05 ). The expression levels of HMGB1 - mRNA in normal group, CRS I group and CRS U group were (0. 81 ± 0. 31),(2. 23 ±0. 35) and (1.61 ±0. 42), respectively, the expression levels in both CRS groups were higher than those in normal group, and the expression level in CRS I group was the highest (P <0.05). The results of Western blot showed that the expression of HMGB1, Beclin - 1 and LC3 in CRS I type were (1.69 ±0.21 ),( 1.89 ±0. 35 ) and ( 1.94 ±0. 28), respectively, which was higher than those in CRS II type (1.31 ±0. 29),(1.39 ±0. 29),( 1.34 ±0.31)(P <0.05), and the expressions in both CRS groups were significantly higher than those in normal group (0. 61 ±0. 23 ),(0. 64 ± 0. 12),(0. 51 ± 0. 22), the difference was statistically significant (P < 0. 05 ). Conclusion The expression of miR - 141 in serum of CRS I patients is decreased, which is related to the fact that miR - 141 affects the level of autophagy by affecting the level of HMGB1, which leads to the occurrence and development of CRS, so miR - 141 may be a potential therapeutic target for CRS.