虎杖苷对成纤维细胞生物学特性的影响

Effects of polydatin on the biological features of cultured fibroblasts

背景:虎杖多年来一直是烧伤、创伤创面愈合治疗方剂中的一味主药,因成分复杂,具体药理作用难以进一步研究,对于虎杖苷促愈合作用目前未见文献报道。目的:分析不同浓度虎杖苷对成纤维细胞生物学特性的影响。方法:取烧伤后行瘢痕切除植皮4例患者剩余小中厚皮片,原代培养人成纤维细胞。用含有10-6,10-5,10-4,10-3,10-2mol/L不同浓度虎杖苷培养液作用第2代人成纤维细胞,未加虎杖苷的培养液作为对照。MTT法检测细胞增殖情况;流式细胞仪检测细胞周期及凋亡;ELISA法检测上清中纤维结合蛋白、Ⅰ型胶原蛋白、Ⅰ型胶原蛋白的表达情况。结果与结论:10-5、10-4mol/L组促进成纤维细胞增殖最明显,10-2mol/L组吸光度值显著下降,细胞生长受抑制。10-3mol/L组G1期细胞大幅度下降,细胞有S期阻滞现象。10-2mol/L组有明确的促凋亡作用。10-2mol/L组上清中Ⅰ、Ⅰ型胶原蛋白较其他各组显著增高(P<0.05);纤维结合蛋白较对照组显著增高(P<0.05),较其他各组有所下降(P<0.05)。说明低浓度虎杖苷有促进成纤维细胞增殖、保护细胞免于凋亡及促进纤维结合蛋白的表达与合成分泌的作用,促进成纤维细胞增殖的最适浓度应为10-5～10-4mol/L。

BACKGROUND： Polydatin has been widely used for treatment of burns and trauma. For its complicated composition, it is difficult to further study the specific pharmacological effects of polydatin. There is no study about local treatment of wound healing with polydatin. OBJECTIVE： To investigate the effects of polydatin with different concentrations on the biological features of cultured fibroblasts. METHODS： Fibroblasts were obtained by the explants culture of human skin and subcultured from four patients treated with scar excision skin grafting after burns. The second passage fibroblasts were cultured in culture media with varied doses of polydatin （10-6, 10-5, 10-4, 10-~, 10.2 mol/L respectively）. Cultured media without polydatin was as control. The proliferation of cultured fibroblasts in vitro was detected by MTT test. The cellular cycle and the apoptosis percent were measured by flow cytometry. ELISA method was used to detect the expression of secreted collagen Ⅰ , collagen Ⅲ and fibronectin in supernatant. RESULTS AND CONCLUSION： Polydatin with the concentration of 105 mol/L and 10 mol/L could enhance fibroblast proliferation in vitro. The absorbance value in 10.2 mol/L polydatin group was decreased and the cell growth was inhibited. Polydatin with the concentration of 103 mol/L could block the cellular cycle in S phase. Polydatin with the concentration of 10-2 mol/L could induce cells to apoptosis. The expressions of collagen Ⅰ , collagen Ⅲ were significantly increased in 10-2 mol/L polydatin group when compared with other groups （P 〈 0.05）; the expression of fibronectin was significantly increased in 10.2 mol/L polydatin group when compared with control group （P 〈 0.05）, and when compared with other polydatin groups, the expression of fibronectin was decreased （P 〈 0.05）. Low concentration of polydation could promote fibroblast proliferation and protect the cells from apoptosis, and it could also improve the expression, synthesis and secretion of fibronectin. To fibroblasts, the optimal concentration of polydatin is 10-5 mol/L and 10^-4 mol/L.