高糖对大鼠腹膜间皮细胞分泌细胞因子的影响及苦参碱的干预作用

摘要

目的：研究高糖刺激下大鼠腹膜间皮细胞分泌肿瘤坏死因子α（TNF‐α）和转化生长因子β1（TGF‐β1）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＝15，A组）；低糖苦参碱组（2．5％葡萄糖，n＝15，B组）；4．25％葡萄糖组（n＝15，C组）；高糖苦参碱组（4．25％葡萄糖，n＝15，D组）；对照组（n＝15，E组，仅加不含血清的DM EM‐F12 BM SC培养基），分别于第24、48、72小时收集培养上清液，采用ELISA法测定各组培养上清液中TNF‐α和TGF‐β1浓度的变化，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E组和B组腹膜间皮细胞小，呈卵圆形或圆形，生长较为密集。A、C、D组腹膜间皮细胞大、呈梭形或多边形，分布较为稀疏；在A组，培养上清液中TNF‐α和TGF‐β1水平较E组无明显统计学变化（P＞0．05）；而在C组，培养上清液中TNF‐α和TGF‐β1水平较E组增加明显，差异有统计学意义（P＜0．05）。在B组和D组中，PMCs分泌TNF‐α和TGF‐β1水平B组较A组减少，差异无统计学意义（P＞0．05）；D组较C组明显下降，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TNF‐α和TGF‐β1，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 tumor necrosis factor‐α(TNF‐α)and transforming growth factor‐β1 (TGF‐β1 ) overexpression in rat peritoneal mesothelial cells(PMCs) ,and to ex‐plore the possible mechanism in the peritoneal fibrosis .Methods ology The rat PMCs were incubated in vitro ,and then assigned in‐to 5 groups on the basis of the added drug concentrations :2 .5% glucose group (n=15 ,A);Glucose + 2 .5% matrine group (n=15 ,B);4 .25% glucose group (n=15 ,C);Glucose + 4 .25% matrine group (n=15 ,D);Blank control group (n=15 ,control group , added the DMEM/F12 medium) .The levels of TNF‐αand TGF‐β1 in the supernatants were measured by ELISA method at the dif‐ferent time (24h ,48h ,72h) ,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 TNF‐αand TGF‐β1 in the A group was lightly higher than that of the control group ,but the level in the C group was significantly higher than that of the control group (P<0 .05) .As compared with the A group ,The level of TNF‐α and TGF‐β1 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 TNF‐αand TGF‐β1 induced by high glucose ,and then protect the PMCs .