高糖对大鼠腹膜间皮细胞分泌FOXP3mRNA的影响及苦参碱的干预作用

Effect of Matrine on High Glucose-induced FOXP3mRNA Overexpression in Rat Peritoneal Mesothelial Cells

目的:研究高糖刺激下大鼠腹膜间皮细胞分泌叉头状螺旋转录因子(FOXP3mRNA)及苦参碱的干预作用,探讨腹膜纤维化发生机制。方法:通过体外培养大鼠腹膜间皮细胞(PMCs),依据加入药物浓度分成5组:2.5%葡萄糖组(n=10,A);2.5%葡萄糖+苦参碱组(n=10,B);4.25%葡萄糖组(n=10,C);4.25%葡萄糖+苦参碱组(n=10,D);空白对照组(仅加不含血清的DMEM/F12培养基),分别于第24 h、48 h、72 h收集培养上清液,采用RT-PCR检测FOXP3mRNA的表达,同时观察各组大鼠腹膜间皮细胞光镜下的表现。结果:光镜下,可见对照组和B组腹膜间皮细胞小,呈圆形、梭形或不规则形,生长密集。A、C、D组腹膜间皮细胞大、呈梭形、分布较稀疏;在含2.5%高糖A组,FOXP3mRNA的表达相对水平较对照组增加,差异有统计学意义(P<0.05);而在4.25%高糖C组,FOXP3mRNA的表达较对照组增加明显,差异有统计学意义(P<0.01)。在含苦参碱溶液的B组和D组中,PMCs表达FOXP3mRNA的相对水平B组较A组减少,但差异无统计学意义(P>0.05);D组较C组下降明显,差异有统计学意义(P<0.05)。结论:苦参碱能拮抗高糖致大鼠腹膜间皮细胞分泌FOXP3mRNA,从而保护腹膜间皮细胞。

Objective： This study was designed to investigate the effect of matrine on high glucose-induced Forkhead box protein 3（ FOXP3mRNA） overexpression in rat peritoneal mesothelial cells（ PMCs）,and to explore the possible mechanism in the peritoneal fibrosis.Methods： The rat PMCs were incubated in vitro,and then assigned into 5 groups on the basis of the added drug concentrations： 2.5% glucose group（ n = 10,A）; Glucose ＋ 2.5% matrine group（ n = 10,B）; 4.25% glucose group（ n = 10,C）; Glucose ＋ 4.25% matrine group（ n = 10,D）; Blank control group（ added the DMEM / F12 medium）.The levels of FOXP3 mRNA in the supernatants were measured by RT-PCR method at the different time（ 24 h、48 h、72 h）,and observed the changes of PMCs under the microscope.Results： Compared with the groups of A,C,and D,the PMCs in the control group and B group were small,round,spindle,irregular or intensive.The expression of FOXP3mRNA in the A group was lightly higher than that of the control group（ P〈0.05）,but the level in the C group was significantly higher than that of the control group（ P〈0.01）.As compared with the A group,The level of FOXP3 mRNA expressed in the B group were decreased（ P〈0.05）,but the lever in the D group was significantly lower than that of the C group（ P〈0.05）.Conclusion： Matrine might be helpful to resist the overexpression of FOXP3 mRNA induced by high glucose,and then protect the PMCs.