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目的研究结缔组织生长因子(CTGF)及其抗体对人胃成纤维细胞增殖、移行、合成细胞外基质(ECM)的影响。方法 (1)应用四唑盐(MTT)比色法检测不同浓度的CTGF(5、20、50、100 ng/ml)及其抗体(0.1、0.5、1μg/ml)对人胃成纤维细胞增殖的影响。(2)应用羟脯氨酸法检测不同浓度的CTGF及其抗体对人胃成纤维细胞分泌胶原蛋白的影响。(3)应用Transwell小室法检测不同浓度的CTGF及其抗体对人胃成纤维细胞迁移能力的影响。结果5～10 ng/ml CTGF可剂量依赖性地促进入胃成纤维细胞增殖;20～100 ng/ml CTGF可剂量依赖性地促进人胃成纤维细胞分泌胶原蛋白;20～100 ng/ml CTGF可剂量依赖性地促进人胃成纤维细胞迁移。另一方面,0.1～1μg/ml CTGF抗体可剂量依赖性地抑制人胃成纤维细胞增殖;0.5、1μg/ml CTGF抗体可剂量依赖性地抑制人胃成纤维细胞分泌胶原蛋白;0.5、1μg/ml ETGF抗体可剂量依赖性地抑制人胃成纤维细胞迁移。以上各实验组与相应对照组比较均有显著性差异(P<0.05)。结论一定浓度范围内的CTGF能剂量依赖性地促进人胃成纤维细胞增殖、分泌胶原蛋白及迁移,这表明CTGF参与了胃硬癌间质纤维化进程,在间质纤维化进程的各环节发挥重要作用。而一定浓度范围内的CTGF抗体可剂量依赖性地抑制这些效应,这为胃硬癌的临床治疗提供了新的思路。
Objective To study the effects of connective tissue growth factor (CTGF) and its antibody on proliferation, migration and synthesis of extracellular matrix (ECM) in human gastric fibroblasts. Methods (1) Tetramethylpyrazine (MTT) colorimetric assay was used to detect the proliferation of human gastric fibroblasts with different concentrations of CTGF (5,20,50,100 ng / ml) and their antibodies (0.1,0.5,1 μg / ml) Impact. (2) The effects of different concentrations of CTGF and its antibody on the secretion of collagen by human gastric fibroblasts were detected by hydroxyproline method. (3) Transwell chamber method was used to detect the effect of different concentrations of CTGF and its antibody on the migration of human gastric fibroblasts. Results CTGF 5 ~ 10 ng / ml promoted the proliferation of gastric fibroblast in a dose - dependent manner. CTGF 20 ~ 100 ng / ml promoted the secretion of collagen in human gastric fibroblast in a dose - dependent manner. CTGF 20 ~ 100 ng / ml It can promote the migration of human gastric fibroblasts in a dose-dependent manner. On the other hand, 0.1 ~ 1μg / ml CTGF could inhibit the proliferation of human gastric fibroblasts in a dose-dependent manner. 0.5,1μg / ml CTGF could inhibit the secretion of collagen in human gastric fibroblasts in a dose- ml ETGF antibody can inhibit the migration of human gastric fibroblast in a dose-dependent manner. The above experimental groups and the corresponding control group were significantly different (P <0.05). Conclusion CTGF in a certain concentration range can promote the proliferation, secretion of collagen and migration of human gastric fibroblast in a dose-dependent manner, which indicates that CTGF is involved in the process of interstitial fibrosis in gastric cancer and plays a role in the process of interstitial fibrosis Important role. However, a certain concentration range of CTGF antibody dose-dependently inhibit these effects, which provides a new idea for the clinical treatment of gastric cancer.