血必净注射液在内皮祖细胞修复脂多糖诱导的肾内皮细胞损伤中的作用及机制

Role and mechanism of Xuebijing injection in the repair of lipopolysaccharide-induced renal endothelial cell injury by endothelial progenitor cells

目的探究血必净注射液在内皮祖细胞修复脂多糖诱导的肾内皮细胞损伤中的作用及机制。方法提取并鉴定大鼠内皮祖细胞和肾脏微血管内皮细胞(RMEC),收集内皮祖细胞的培养上清作为条件培养基(CM)。取肾脏微血管内皮细胞,分为6组,空白组正常培养,模型组给予脂多糖(LPS)培养,CM+LPS组、CM+LPS+血必净12.5 mg/mL组、CM+LPS+血必净25 mg/mL组、CM+LPS+血必净50 mg/mL组给予相应干预。MTT法检测各组细胞增殖情况,Transwell小室法检测细胞迁移能力,小管形成实验检测体外血管新生能力,流式细胞术检测细胞凋亡情况,qRT-PCR法检测血管内皮生长因子(VEGF)和碱性成纤维生长因子(bFGF)表达情况,Western blot法检测NF-κB表达情况。结果模型组细胞增殖检测OD值及VEGF、bFGF相对表达量均明显低于空白组,细胞迁移数、小管形成数均明显少于空白组,细胞凋亡率及NF-κB相对表达量明显高于空白组,差异均有统计学意义(P均<0.05);CM+LPS组及血必净各组细胞增殖检测OD值及VEGF、bFGF相对表达量均明显高于模型组,细胞迁移数、小管形成数均明显多于模型组,细胞凋亡率及NF-κB相对表达量明显低于模型组,且血必净各组上述指标改善情况均明显优于CM+LPS组(P均<0.05)。结论血必净注射液可通过抑制NF-κB信号通路而促进内皮祖细胞修复脂多糖诱导的肾内皮细胞损伤。

Objective It is to investigate the role and mechanism Xuebijing injection in the repair of lipopolysaccharide-induced renal endothelial cell injury by endothelial progenitor cells.Methods The rat endothelial progenitor cells and renal microvascular endothelial cells(RMEC)were extracted and identified,and the culture supernatant of the endothelial progenitor cells was collected as the conditioned medium(CM).The renal microvascular endothelial cells were divided into 6 groups.The blank group was cultured normally,and the model group was cultured with lipopolysaccharide(LPS),CM+LPS group,CM+LPS+Xuebijing 12.5 mg/mL group,CM+LPS+Xuebijing 25 mg/mL group,CM+LPS+Xuebijing 50 mg/mL group were given corresponding intervention.The cell proliferation in each group was detected by MTT method,the cell migration ability was detected by Transwell chamber method,the angiogenesis ability in vitro was detected by tubule formation test,the cell apoptosis was detected by flow cytometry,the expression of vascular endothelial growth factor(VEGF)and basic fibroblast growth factor(bFGF)was detected by qRT-PCR method,the expression of NF-κB was detected by Western blot.Results The cell proliferation detection OD value and the relative expression of VEGF and bFGF in the model group were significantly lower than those in the blank group,the number of cell migration and tubule formation were significantly less than those in the blank group,the apoptosis rate and the relative expression of NF-κB were significantly higher than those in the blank group,the differences were statistically significant(all P<0.05);the cell proliferation detection OD value and the relative expression of VEGF and bFGF in the CM+LPS group and each Xuebijing group were significantly higher than those in the model group,the number of cell migration and tubule formation were significantly more than those in the model group,the apoptosis rate and the relative expression of NF-κB were significantly lower than those in the model group,and the improvement of the above indicators in each group of Xuebijing was significantly better than that of the CM+LPS group,the differences were all significant(all P<0.05).Conclusion Xuebijing injection can promote endothelial progenitor cells to repair lipopolysaccharide-induced renal endothelial cell injury by inhibiting NF-κB signaling pathway.