木犀草素通过调节细胞凋亡和炎症发挥对内毒素性所致急性肾损伤的保护作用

Protective effects of luteolin on endotoxic-induced acute kidney injury by modulating apoptosis and inflammation

目的探讨木犀草素对急性肾损伤的保护作用。方法通过脂多糖(LPS构建小鼠内毒素性所致的急性肾损伤模型,使用木犀草素对肾损伤小鼠进行治疗;给小鼠尾静脉注射miR-30c-5p antagomir以敲低miR-30c-5p的表达。动物分组如下:正常对照组、LPS组、木犀草素治疗组(LPS+Lu组)、木犀草素治疗的阴性对照组(LPS+DMSO组)、miR-30c-5p表达敲低组(antagomir 30c-5p组)、miR-30c-5p表达敲低对照组(antagomir NC组)。采用自动化学分析仪测定小鼠血清肌酐和尿素氮水平,通过HE染色观察肾组织病理损伤。采用ELISA试剂盒检测小鼠血清胱抑素C(Cys-C)、肿瘤坏死因子(TNF)-α、白细胞介素(IL)-6、IL-4水平,通过TUNEL染色和Western blot检测细胞凋亡。结果与正常对照组相比,LPS组小鼠急性肾损伤病理情况严重,血清肌酐、尿素氮和Cys-C、促炎因子IL-6、TNF-α、促凋亡因子B淋巴细胞瘤-2基因(Bcl-2)相关X蛋白(Bax)的表达水平显著升高(P<0.05),而抗炎因子IL-4、抗凋亡因子Bcl-2、miR-30c-5p水平显著下降(P<0.05)。与LPS组相比,LPS+Lu组肾损伤病理情况减轻,血清肌酐、尿素氮和Cys-C、IL-6、TNF-α、Bax的表达水平显著降低,而IL-4、Bcl-2、miR-30c-5p水平显著上升(P<0.05)。与antagomir NC组小鼠相比,antagomir 30c-5p组小鼠促炎因子水平及细胞凋亡率显著上升(P<0.05)。结论木犀草素通过上调miR-30c-5p表达抑制炎症和细胞凋亡从而保护急性肾损伤小鼠。

Objective To explore the protective effect of luteolin on acute kidney injury.Methods Mice with endotoxin-induced acute kidney injury model were constructed by lipopolysaccharide(LPS).Kidney-injured mice were treated with luteolin and given tail vein injection of miR-30c-5p antagomir to knock down the expression of miR-30c-5p.Animals were grouped as follows:normal control group,LPS group,lignocaine treated group(LPS+Lu group),negative control group for lignocaine treatment(LPS+DMSO group),miR-30c-5p expression knockdown group(antagomir 30c-5p group)and miR-30c-5p expression knockdown control group(antagomir NC group).Serum creatinine(SCr)and blood urea nitrogen(BUN)were measured in mice by taking automated chemical analyzer,and renal histopathological damage was observed by HE staining.Serum Cystatin C(Cys-C),inflammatory factors tumor necrosis factor(TNF)-α,interleukin(IL)-6 and IL-4 levels were measured in mice using ELISA kits,and apoptosis was detected by TUNEL staining and Western blot.Results Compared with the normal control group,acute kidney injury pathology was more severe in the LPS group,with significantly higher expression levels of SCr,BUN and serum Cys-C,pro-inflammatory factor IL-6,TNF-α,and pro-apoptotic factor Bax protein expression level,and expression levels of anti-inflammatory factor IL-4,anti-apoptotic factor Bcl-2,and miR-30c-5p expression levels were significantly decreased(P<0.05).Compared with the LPS group,the pathology of kidney injury was reduced in the LPS+Lu group,and the expression levels of SCr,BUN and Cys-C,IL-6,TNF-α,and Bax were significantly reduced,while the levels of IL-4,Bcl-2,and miR-30c-5p were significantly increased(P<0.05).Compared with mice in the antagomir NC group,antagomir 30c-5p group mice owned more severe inflammation and apoptosis(P<0.05).Conclusion Luteolin protected mice with acute kidney injury by upregulating miR-30c-5p expression to inhibit inflammation and apoptosis.