Effects of Dragon’s blood extracts on fibroblast proliferation and procollagen type III

doi:10.3969/j.issn.2095-4344.2014.46.012

Abstract

Abstract:

BACKGROUND: Dragon’s blood is the main ingredient of traditional medicine prescription for promoting granulation, which has been used in clinical treatment of a variety of refractory wounds and achieved the exact effects. But the Dragon’s blood effect on collagen secretion from normal fibroblasts has not been reported.

OBJECTIVE: To investigate the effects of Dragon’s blood extract on the proliferation and secret function of fibroblasts in vitro.

METHODS: Dragon’s blood was extracted by extracts chloroform, acetoacetic ester, and alcohol in turn. Normal human fibroblasts were respectively cultured in Dragon’s blood extracts of chloroform, acetoacetic ester, and alcohol, DMEM containing 1% dimethyl sulfoxide, and normal culture medium. Then, the fibroblasts were cultured in vitro in different media containing gradient dilutions of Dragon’s blood extracts (0.002, 0.02, 0.2, 2, 20 g/L), which was followed by cell proliferation determination assessed with MTT assay. Under the optimal concentration, the cell growth curves were drawn and the flow cytometry was used to determine the changes of cell cycle. The concentration of procollagen type III in the supernatant of the fibroblast culture systems was measured by radioimmunoassay.

RESULTS AND CONCLUSION: 0.2 g/L-2 g/L dilution of Dragon’s blood extracted by acetoacetic ester enhanced the proliferation of fibroblasts in a dose-dependent manner. The 2 g/L was the optimal dilution of Dragon’s blood extracted by acetoacetic ester, and it increased the ratio of S cells in cell cycle than control group and decreased procollagen type III. These findings indicate that Dragon’s blood acetoacetic ester extract can improve the proliferation of cultured human fibroblasts in vitro, and decrease the secretion of procollagen type III of fibroblasts, and it can be beneficial to improve wound healing and inhibit hypertrophic scar.

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

全文链接：

Key words: fibroblasts, drugs, chinese herbal, plant extracts, cell proliferation

CLC Number:

R318

Li Dan, Hui Rui, Hu Yong-wu, Han Yan, Guo Shu-zhong. Effects of Dragon’s blood extracts on fibroblast proliferation and procollagen type III [J]. Chinese Journal of Tissue Engineering Research, 2014, 18(46): 7437-7441.

Figures/Tables 3

2.1 血竭提取物对成纤维细胞增殖的影响及最适浓度筛选氯仿提取组及乙醇提取组在各种浓度条件下，对成纤维细胞增殖的影响与对照组相比，差异均无显著性意义 (P > 0.05)。乙酸乙酯提取物在浓度为0.2-2 g/L时均有不同程度促进成纤维细胞增殖的作用，并且呈剂量依赖关系， 2 g/L时促进成纤维细胞作用最显著(P < 0.05)，故2 g/L血竭乙酸乙酯提取物为促进成纤维细胞增殖的最适浓度。含1%二甲基亚砜组与对照组无显著差异(P > 0.05；图1)。因此2 g/L乙酸乙酯提取物为最适合成纤维细胞生长的一组血竭提取物，后面的实验仅探讨其与对照组的差异。 2.2 血竭提取物对成纤维细胞形态的影响光学显微镜下可见，在质量浓度为2 g/L的乙酸乙酯提取物培养24 h后，成纤维细胞的细胞融合现象较对照组明显，对照组仅部分成纤维细胞散在融合(图2)。 2.3 血竭提取物最佳浓度条件下的成纤维细胞生长曲线成纤维细胞在血竭乙酸乙酯提取物最佳浓度2 g/L条件下培养，可见培养8 h时其吸光度值明显高于对照组(P < 0.05)，且随时间的延长而逐渐增大，在24 h时2组的吸光度值均达到峰值(P < 0.05)，而后2组的吸光度值逐渐下降，但2 g/L血竭乙酸乙酯提取物组细胞的吸光度值仍明显高于对照组(P < 0.05；图3)。 2.4 血竭提取物对成纤维细胞细胞周期的作用流式细胞术检测到，培养24 h后，2 g/L血竭乙酸乙酯提取物组成纤维细胞处于G1期的比例明显低于对照组(P < 0.05)，但S期的细胞比例则明显高于对照组(P < 0.05)；见表1。 2.5 血竭提取物对成纤维细胞分泌Ⅲ型前胶原的影响在培养0-72 h时，2 g/L血竭乙酸乙酯提取物组与对照组成纤维细胞中Ⅲ型前胶原水平均呈持续递增的趋势，且其中 2 g/L血竭乙酸乙酯提取物组成纤维细胞中Ⅲ型前胶原水平在24-72 h低于对照组(P < 0.05)；见表2。"

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