论文部分内容阅读
目的:探讨3种中药提取物对体外培养人真皮成纤维细胞增殖及胶原合成的影响。方法:采用组织块法分离培养人真皮成纤维细胞,进行波形蛋白免疫组化鉴定。MTT法测定3种中药提取物作用后成纤维细胞的增殖能力,消化法检测培养液中羟脯氨酸的含量变化。结果:培养的细胞为典型的成纤维细胞形态,波形蛋白免疫组化呈阳性;冻存前后的人真皮成纤维细胞均具有较强的增殖能力;MTT检测结果显示培养72 h后,0.312 5~5.000 0 mg·ml~(-1)芦荟凝胶提取物、0.156 3~2.500 0 mg·ml~(-1)白芨提取物、0.075 0 mg·ml~(-1)丹参多酚酸盐,具有明显促进人成纤维细胞增殖作用(P<0.05,P<0.01),且培养上清液中羟脯氨酸含量高于空白对照组(P<0.01,P<0.05)。丹参多酚酸盐对成纤维细胞增殖的影响与时间和浓度关系密切,随着培养时间延长,高浓度药物组对成纤维细胞由促进变为抑制作用。结论:芦荟、白芨、丹参提取物可通过促进人真皮成纤维细胞增殖以及羟脯氨酸等细胞外基质的分泌加速创面愈合。
Objective: To investigate the effects of three kinds of traditional Chinese medicine extracts on the proliferation and collagen synthesis of cultured human dermal fibroblasts in vitro. Methods: Human dermal fibroblasts were isolated and cultured by tissue block method and identified by vimentin immunohistochemistry. MTT assay was used to determine the proliferation of fibroblasts after three kinds of traditional Chinese medicine extracts. The content of hydroxyproline in culture solution was determined by digestion method. RESULTS: The cultured cells were typical fibroblast morphology with immunohistochemistry of vimentin. Human dermal fibroblasts before and after cryopreservation had strong proliferative ability. The results of MTT assay showed that after cultured for 72 h, 5.000 0 mg · ml -1 aloe vera gel extract, 0.156 3 ~ 2.500 0 mg · ml -1 whitefly extract, 0.075 0 mg · ml -1 salvianolic acid salt, with (P <0.05, P <0.01), and the content of hydroxyproline in the culture supernatant was higher than that of the blank control group (P <0.01, P <0.05). The effect of Salviae miltiorrhizae polyphenols on the proliferation of fibroblasts was closely related to the time and concentration. With the prolongation of culture time, the high concentration of drug group could inhibit the fibroblasts from promoting. Conclusion: Aloe, Paeonia lactiflora and Salvia miltiorrhiza extract can accelerate wound healing by promoting the proliferation of human dermal fibroblasts and secretion of extracellular matrix such as hydroxyproline.