荜茇酰胺通过miR-132-3p调控FOXO1参与脓毒症肾损伤研究

目的：探究荜茇酰胺对脓毒症肾损伤的作用及其可能机制。方法：使用盲肠结扎穿刺法(CLP)构建小鼠脓毒症模型。使用苏木素-伊红染色(HE)观察小鼠肾组织形态。使用Luminex LX100系统检测小鼠血清中肾损伤标志物的表达情况。使用酶联免疫反应(ELISA)检测小鼠肾组织中炎症因子水平。使用原位缺口末端标记法(TUNEL)检测小鼠肾组织细胞凋亡水平。使用脂多糖(LPS)构建脓毒症细胞模型。使用细胞计数试剂盒8(CCK8)法检测细胞活性。使用流式细胞术检测细胞凋亡水平。使用qRT-PCR检测miR-132-3p表达量。使用蛋白免疫印迹法(WB)检测FOXO1和凋亡相关蛋白的表达情况。使用双荧光素酶实验检测miR-132-3p与FOXO1靶向结合关系。结果：小鼠体内实验中,与假手术组相比,脓毒症模型组小鼠外周血中肾损伤标志物表达上升,肾损伤显著,肾组织中炎症因子表达显著上升,肾组织细胞凋亡水平上升,miR-132-3p表达显著上升,FOXO1蛋白表达下降；与脓毒症模型组比,荜茇酰胺给药组小鼠外周血中肾损伤标志物表达下降,肾损伤有所缓解,肾组织中炎症因子表达显著下降,肾组织细胞凋亡水平下降,miR-132-3p表达显著下降,FOXO1蛋白表达上升。在体外细胞模型中,荜茇酰胺能显著提高LPS刺激下肾小管细胞活性,抑制凋亡水平,并且过表达miR-132-3p会抑制荜茇酰胺对LPS刺激下细胞的影响。通过双荧光素酶实验在体外细胞中证实了miR-132-3p对FOXO1的靶向调控作用,且过表达miR-132-3p时FOXO1表达显著下调,抑制miR-132-3p表达时FOXO1表达显著上调。结论：荜茇酰胺可能通过miR-132-3p调控FOXO1的表达从而缓解脓毒症肾损伤。

Piperlongumine regulates FOXO1 through miR-132-3p to participate in septic kidney injury

Objective To evaluate the effect of piperlongumine on renal injury in sepsis and its possible mechanism. Methods: Mouse sepsis model was established by cecal ligation and puncture (CLP). Hematoxylin-eosin staining (HE) was used to observe the morphology of mouse kidney tissue. Luminex LX100 system was used to detect the expression of kidney injury markers in mouse serum. The levels of inflammatory factors in mouse kidney tissue were detected by enzyme-linked immune reaction (ELISA). TUNEL was used to detect apoptosis in mouse renal tissue. A cell model of sepsis was constructed using LPS. Cell activity was detected by CCK8 assay. The apoptosis level was detected by flow cytometry. The expression of miR-132-3p was detected by qRT-PCR. The expressions of FOXO1 and apoptosis-related proteins were detected by WB. Dual luciferase assay was used to detect the targeting binding relationship between miR-132-3p and FOXO1.