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纸质出版:2010
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徐家云,樊小军,常洁,马沙,高翔,田祥银,董少卿,王青苗.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[J].陆军军医大学学报(原第三军医大学学报),2010,32(13):1446-1448.
Xu Jiayun,Fan Xiaojun,Chang Jie,Ma Sha,Gao Xiang,Tian Xiangyin,Dong Shaoqing,Wang Qingmiao.Effect of chitosan on expression of transforming growth factor-β1 in peritoneum mesothelial cells stimulated by high-glucose peritoneal dialysate[J].J Amry Med Univ (J Third Mil Med Univ),2010,32(13):1446-1448.
徐家云,樊小军,常洁,马沙,高翔,田祥银,董少卿,王青苗.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[J].陆军军医大学学报(原第三军医大学学报),2010,32(13):1446-1448. DOI:
Xu Jiayun,Fan Xiaojun,Chang Jie,Ma Sha,Gao Xiang,Tian Xiangyin,Dong Shaoqing,Wang Qingmiao.Effect of chitosan on expression of transforming growth factor-β1 in peritoneum mesothelial cells stimulated by high-glucose peritoneal dialysate[J].J Amry Med Univ (J Third Mil Med Univ),2010,32(13):1446-1448. DOI:
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1.2 mg/L;D组:DMEM/F12培养基中加入4.25%葡萄糖透析液+几丁糖0.4 mg/L。培养24 h收集细胞提取RNA,48 h收集细胞提取蛋白,RT-PCR检测腹膜间皮细胞TGF-β1 mRNA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-β1 mRNA和蛋白的表达均上升,与0 h比较,TGF-β1 mRNA和蛋白的表达分别于24、48 h时达到高峰, A、B两组比较差异有统计学意义(P0.05);②24 h时,几丁糖使TGF-β1 mRNA的表达下降明显(P0.05),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
P0.01)。48 h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(P0.05),且抑制程度与几丁糖浓度呈正相关(r=0.831
P0.05)。 结论 ① 高糖腹透液可刺激RPMC TGF-β1 mRNA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 expression 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.
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