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目的观察exendin-4对大鼠肾小球系膜细胞单核细胞趋化蛋白-1(MCP-1)、纤维连接蛋白(FN)表达的影响。方法试验分组:对照组、肿瘤坏死因子-α(TNF-α)组(10 ng/ml)、TNF-α(10 ng/ml)+E1组(1 nmol/L exendin-4)、TNF-α(10 ng/ml)+E5组(5 nmol/L exendin-4)、TNF-α(10 ng/ml)+E10组(10 nmol/L exendin-4)。收集24 h细胞,提取RNA,采用实时荧光定量检测细胞内MCP-1 mRNA的表达;分别在24、48 h收集各组细胞培养上清液,采用ELISA检测培养细胞上清中MCP-1、FN的浓度。结果孵育24 h时,TNF-α组MCP-1 mRNA和细胞培养上清MCP-1、FN表达较对照组明显增加(P<0.01);与TNF-α组比较,TNF-α+E5组、TNF-α+E10组细胞内MCP-1 mRNA和培养上清内MCP-1、FN含量均明显降低;孵育48 h时,TNF-α组细胞培养上清MCP-1、FN表达较对照组明显增加(P<0.01);与TNF-α组比较,TNF-α+E1组、TNF-α+E5组、TNF-α+E10组细胞培养上清内MCP-1、FN含量均明显降低;并且随着时间的延长和exendin-4浓度的增加,培养上清内MCP-1、FN含量降低更加明显。结论 Exendin-4可明显抑制TNF-α诱导的系膜细胞MCP-1 mRNA的表达,并以剂量和时间依赖的方式抑制TNF-α诱导的系膜细胞培养上清内MCP-1、FN的含量,提示exendin-4可能会对肾脏具有一定保护作用。
Objective To investigate the effect of exendin-4 on the expression of monocyte chemoattractant protein-1 (MCP-1) and fibronectin (FN) in rat mesangial cells. Methods The experimental groups were divided into control group, TNF-α group (10 ng / ml), TNF-α (10 ng / ml) group and E1 group (1 nmol / L exendin-4) (10 ng / ml) + E5 group (5 nmol / L exendin-4), TNF-α (10 ng / ml) + E10 group (10 nmol / L exendin-4). The cells were collected for 24 h, RNA was extracted and the expression of MCP-1 mRNA was detected by real-time fluorescence quantitative detection. The supernatant of each group was collected at 24,48 h. The expression of MCP-1, FN concentration. Results Compared with TNF-α group, the expression of MCP-1 mRNA and MCP-1, FN in TNF-α group and TNF-α group were significantly increased compared with control group (P <0.01) The levels of MCP-1 mRNA and MCP-1 and FN in the TNF-α + E10 group were significantly lower than those in the control group at 48 h after incubation (P <0.01). Compared with TNF-α group, the contents of MCP-1 and FN in TNF-α + E1 group, TNF-α + E5 group and TNF-α + E10 group were significantly decreased; With the prolongation of time and the increase of exendin-4 concentration, the decrease of MCP-1 and FN in culture supernatant was more obvious. Conclusions Exendin-4 can significantly inhibit the expression of MCP-1 mRNA in mesangial cells induced by TNF-α and inhibit the content of MCP-1 and FN in the supernatant of mesangial cells induced by TNF-α in a dose-and time-dependent manner , Suggesting exendin-4 may have some protective effect on the kidneys.