几丁糖对高糖腹透液刺激大鼠腹膜间皮细胞TGF-β1表达的影响  

Effect of chitosan on expression of transforming growth factor-β1 in peritoneum mesothelial cells stimulated by high-glucose peritoneal dialysate

作　　者：徐家云[1] 樊小军[2] 常洁[1] 马沙[1] 高翔[1] 田祥银[1] 董少卿[1] 王青苗[1] 

机构地区：[1]河南科技大学第一附属医院肾内科,河南洛阳471003 [2]西安交通大学医学院电镜室,西安710061

出　　处：《第三军医大学学报》2010年第13期1446-1448,共3页Journal of Third Military Medical University

摘　　要：目的探讨高糖腹透液(4.25%)对大鼠二代腹膜间皮细胞TGF-β1表达的影响及几丁糖(chitosan)的干预作用。方法取第2代鼠腹膜间皮细胞(rat peritoneal mesothelial cells,RPMC)(经细胞形态和免疫组化鉴定),随机分为4组(n=4),A组即对照组:DMEM/F12培养基培养;B组:DMEM/F12培养基中加入4.25%葡萄糖透析液;C组:DMEM/F12培养基中加入4.25%葡萄糖透析液+几丁糖1.2mg/L;D组:DMEM/F12培养基中加入4.25%葡萄糖透析液+几丁糖0.4mg/L。培养24h收集细胞提取RNA,48h收集细胞提取蛋白,RT-PCR检测腹膜间皮细胞TGF-β1mRNA的表达。Western印迹检测TGF-β1的蛋白表达。结果①RPMC经高糖腹透液(4.25%)刺激后,TGF-β1mRNA和蛋白的表达均上升,与0h比较,TGF-β1mRNA和蛋白的表达分别于24、48h时达到高峰,A、B两组比较差异有统计学意义(P<0.05);②24h时,几丁糖使TGF-β1mRNA的表达下降明显(P<0.05),B组分别为A组的2.51倍、C组的2.13倍、D组的1.68倍,D组为C组的1.26倍且抑制程度与几丁糖浓度呈正相关(r=0.948,P<0.01)。48h时,几丁糖使TGF-β1的蛋白水平下降明显,B组分别为A组的1.85倍、C组的1.52倍、D组的1.24倍,D组为C组的1.23倍。C、D两组分别与B组比较差异有统计学意义(P<0.05),且抑制程度与几丁糖浓度呈正相关(r=0.831,P<0.05)。结论①高糖腹透液可刺激RPMCTGF-β1mRNA和蛋白表达上调;②几丁糖可抑制腹膜纤维化的中枢性因子TGF-β1表达的活性。Objective To study the effect of high-glucose peritoneal dialysate （4.25%） on expre-ssion of transforming growth factor-β1（TGF-β1） in peritoneum mesothelial cells （PMC） and the interfering role of chitosan.Methods PMC （second generation）,identified by their morphology and immunohistochemistry,were divided into group A （control group）,group B,group C and group D,which were cultured with DMEM/F12,DMEM/F12 ＋4.25% glucose peritoneal dialysate,DMEM/F12 ＋ 4.25% glucose peritoneal dialysate ＋1.2 mg/L chitosan,and DMEM/F12＋4.25% glucose peritoneal dialysate ＋ 0.4 mg/L chitosan,respectively.RNA and protein were extracted from PMC,respectively,24 and 48 h after culture.Expression of TGF-β1 mRNA and protein in PMC was detected by RT-PCR and Western blot analysis,respectively.Results The expression level of TGF-β1 mRNA and protein in PMC was higher after than before 4.25% glucose stimulation,and reached its peak at 24 and 48 h of culture,in groups A and B,respectively （P0.05）.Chitosan significantly up-regulated the expression of TGF-β1 mRNA and protein in PMC while it significantly down-regulated the expression of TGF-β1 mRNA at 24 h of culture.The expression of TGF-β1 mRNA was 2.51-fold,2.13-fold,1.68-fold higher in group B than in groups A,C,and D （P0.05）.Chitosan significantly decreased the expression of TGF-β1 protein at 48 h of culture.The expression of TGF-β1 protein was 1.85-fold,1.52-fold,1.24-fold lower in group B than in groups A,C,and D （P0.05）.Chitosan inhibited the expression of TGF-β1 protein in a concentration-dependent manner.Conclusion High glucose peritoneal dialysate （4.25%） can up-regulate the expression of TGF-β1 mRNA and protein in PMC.Chitosan can inhibit the expression activity of TGF-β1 during peritoneal fibrosis.

关 键 词：高糖腹透液 腹膜间皮细胞 转化生长因子-Β1 几丁糖 

分 类 号：R459.5[医药卫生—治疗学] R329.5[医药卫生—临床医学]