LncRNA NEAT1与miR-495-3p对重症肺部感染的诊断价值及其对T淋巴细胞凋亡的影响

Diagnostic value of LncRNA NEAT1 and miR-495-3p in severe lung infection and its effect on T lymphocyte apoptosis

目的探讨长链非编码RNA NEAT1(LncRNA NEAT1)与微小RNA-495-3p(miR-495-3p)对重症肺部感染的诊断价值及其对T淋巴细胞凋亡的影响。方法收集95例肺部感染患者为研究对象,分为重症感染组(50例)与非重症感染组(45例),选取40例健康志愿者为对照组。实时荧光定量聚合酶链反应(qRT-PCR)检测NEAT1与miR-495-3p的表达量;采用受试者工作特征(ROC)分析血清NEAT1、miR-495-3p对重症感染的诊断价值。采用免疫磁珠分选外周血T淋巴细胞,分别将si-NC、si-NEAT1转染至T淋巴细胞;流式细胞术检测细胞凋亡率;蛋白免疫印迹法(Western blot)检测B淋巴细胞瘤-2(Bcl-2)、B淋巴细胞瘤-2相关蛋白(Bax)表达量。结果与对照组相比,非重症感染组、重症感染组患者血清和T淋巴细胞中NEAT1的表达水平显著升高(P<0.05),miR-495-3p的表达水平显著降低(P<0.05),非重症感染组与重症感染组间差异亦有统计学意义(P<0.05);ROC曲线分析显示NEAT1在重症感染诊断时敏感度为82.00%,特异度为84.44%,AUC面积为0.816;miR-495-3p在重症感染诊断时敏感度为78.00%,特异度为82.22%,AUC面积为0.820;与对照组相比,非重症感染组与重症感染组T淋巴细胞凋亡率显著升高(P<0.05),Bcl-2蛋白水平显著降低(P<0.05),Bax蛋白水平显著升高(P<0.05),非重症感染组与重症感染组间差异亦有统计学意义(P<0.05);与si-NC组相比,si-NEAT1组T淋巴细胞凋亡率显著降低(P<0.05),Bcl-2蛋白水平显著升高(P<0.05),Bax蛋白水平显著降低(P<0.05),miR-495-3p的表达水平显著升高(P<0.05)。结论重症肺部感染患者血清中NEAT1的表达水平升高,miR-495-3p的表达水平降低,二者对重症肺部感染具有一定诊断价值,沉默NEAT1可能通过上调miR-495-3p表达从而抑制重症肺部感染患者T淋巴细胞凋亡。

Objective To investigate the diagnostic value of LncRNA NEAT1 and miR-495-3p in severe pulmonary infection and its effect on T lymphocyte apoptosis.Methods Ninety-five patients with pulmonary infection were collected as research objects,and were divided into severely infected group(50 cases)and non-severely infected group(45 cases),and 40 healthy volunteers were selected as the control group.qRT-PCR was used to detect the expression of NEAT1 and miR-495-3p.ROC was used to analyze the diagnostic value of serum NEAT1 and miR-495-3p in severe infection.T lymphocytes in peripheral blood were sorted by immunomagnetic beads,and si-NC and si-NEAT1 were transfected into T lymphocytes.Flow cytometry was used to detect the apoptotic rate.Western blot was used to detect the expression of Bcl-2 and Bax.Results Compared with the control group,the expression level of NEAT1 in serum and T lymphocytes of patients with non-severely infected group and severely infected group was significantly increased(P<0.05),and the expression level of miR-495-3p was significantly reduced(P<0.05).The difference between the non-severely infected group and the severely infected group was statistically significant(P<0.05).ROC analysis showed that the sensitivity of NEAT1 was 82.00%,the specificity was 84.44%,and the AUC area was 0.816.The sensitivity of miR-495-3p was 78.00%,the specificity was 82.22%,and the AUC area was 0.820.Compared with the control group,the apoptosis rate of T lymphocytes in non-severely infected group and severely infected group was significantly increased(P<0.05),the protein level of Bcl-2 was significantly reduced(P<0.05),and the protein level of Bax was significantly increased(P<0.05).The difference between the non-severely infected group and the severely infected group was statistically significant(P<0.05).Compared with the si-NC group,the apoptosis rate of T lymphocytes in the si-NEAT1 group was significantly reduced(P<0.05),the protein level of Bcl-2 was significantly increased(P<0.05),the protein level of Bax was significantly reduced(P<0.05),and miR-495-3p expression level was significantly increased(P<0.05).Conclusion The expression level of NEAT1 in the serum of patients with severe lung infection is increased,and the expression level of miR-495-3p is decreased,which has certain diagnostic value for severe lung infection,and silent BEAT1 may inhibit T lymphocyte apoptosis in patients with severe lung infection by upregulating the expression of miR-495-3p.