6-O-羧甲基壳聚糖对人增生性瘢痕成纤维细胞TGF-β1表达的影响(1)
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[摘要]目的：观察不同浓度6-O-羧甲基壳聚糖(6-O-CMC)对体外培养的人增生性瘢痕成纤维细胞(hypertrophic scar fibrob lasts，HSFBs )转化生长因子-β1 (transforming growth factor-beta1，TGF-β1) 蛋白及mRNA表达的影响，在分子水平上探讨其抑制增生性瘢痕生长的作用机制。 方法：体外培养人HSFBs，选取第3～5代细胞进行下一步实验。实验分组采用含不同浓度6-O-CMC(0，100，200，400μg/ml)的DMEM培养液分别进行培养。培养24h后，显微镜下观察各组HSFBs形态及生长情况。收集细胞培养上清，运用双抗体夹心ELISA法检测各组TGF-β1蛋白表达水平。运用荧光定量RT-PCR技术检测各组TGF-β1 mRNA表达水平。 结果：与空白组相比，三个药物剂量组HSFBs TGF-β1蛋白及mRNA的表达均减少(P<0.05)，药物浓度越高表达量越低(P<0.05)，且TGF-β1 蛋白及mRNA表达的变化具有一致性。 结论：6-O-CMC有抑制体外培养的人HSFBs TGF-β1表达的作用，初步探讨了6-O-CMC发挥抑制增生性瘢痕生长的作用机制，为羧甲基壳聚糖类药物的开发利用提供理论依据。
[关键词]6-O-羧甲基壳聚糖；增生性瘢痕成纤维细胞；TGF-β1；ELISA；荧光定量RT-PCR
[中图分类号]R619+.6[文献标识码]A[文章编号]1008-6455(2011)02-0240-03
Effects of 6-O-carboxymethyl-chitosan on the expression of TGF-β1 in human hypertrophic scar fibroblasts
WU Dan1,KUANG Rui-xia2,WANG Zhi-guo2,CHEN Lu2,ZHOU Ming1,HUANG Hong-jie1
(1.Medical College of Qingdao University,Qingdao 266021,Shandong,China； 2.Department of Plastic Surgery,The Affiliated Hospital of Medical College Qingdao University,Qingdao 266013,Shandong, China)
Abstract:ObjectiveTo observe the effects of 6-O-carboxymethyl-chitosan (6-O-CMC) with different concentrations on the expression of transforming growth factor-beta1 (TGF-β1) in human hypertrophic scar fibroblasts (HSFBs) in vitro.MethodsHSFBs were cultured in vitro,and the 3～5 generation cells were chose to the next experiment. The DMEM medium with different concentrations of 6-O-CMC (100,200,400ug/ml) were used in the experimental groups, and the control group without 6-O-CMC. After 24 hours, cell morphology and growth state of HSFBs were observed under the microscope. The expression of TGF-β1 protein was detected by ELISA. The expression of TGF-β1 mRNA was was detected by Real-time RT-PCR. ResultsCompared with the control group, the expressions of HSFBs TGF-β1 protein and mRNA in the other groups all reduced obviously (P<0.05). Concentration and expression were in inverse proportion(P<0.05). The expression of TGF-β1 protein and mRNA are on the same direction ......
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