Hu Feng, Yan Yu, Wang Chu-Wang, Liu Yu, Wang Jing-Jing, Zhou Fang, Zeng Qing-Hai, Zhou Xiao, Chen Jia, Wang Ai-Jun, Zhou Jian-da
Department of Plastic Surgery, the Third Xiangya Hospital of Central South University, Changsha, 410013, China.
Xiangya School of Medicine, Central South University, Changsha, 410013, China.
Chin J Integr Med. 2019 Mar;25(3):203-209. doi: 10.1007/s11655-018-3060-9. Epub 2018 Dec 15.
To investigate the effects of Ganoderma lucidum polysaccharides (GL-PS) on human fibroblasts and skin wound healing in Kunming male mice and to explore the putative molecular mechanism.
Primary human skin fibroblasts were cultured. The viability of fibroblasts treated with 0, 10, 20, 40, 80, and 160 μg/mL of GL-PS, respectively were detected by 3-4,5-dimethyl-2-thiazolyl-2,5-diphenyl-2-Htetrazolium bromide (MTT). The migration ability of fibroblasts treated with 0, 10, 20, and 40 μg/mL of GL-PS were measured by transwell assay. The secretion of the C-terminal peptide of procollagen type I (CICP) and transforming growth factor-β1 (TGF-β1) in the cell supernatant was tested by enzyme-linked immunosorbent assay. The expression of β-catenin was detected by Western blot. Furthermore, the Kunming mouse model with full-layer skin resection trauma was established, and was treated with 10, 20, and 40 mg/mL of GL-PS, respectively as external use. The size of the wound was measured daily, complete healing time in each group was recorded and the percentage of wound contraction was calculated.
Compared with the control group, 10, 20, and 40 μg/mL of GL-PS significantly increased the viability of fibroblasts, promoted the migration ability of fibroblasts, and up-regulated the expressions of CICP and TGF-β1 in fibroblasts (Plt;0.05 or Plt;0.01). The expression of β-catenin in fibroblasts treated with 20 and 40 μg/mL of GL-PS was significantly higher than that of the control group (Plt;0.01). Furthermore, after external use of 10, 20, and 40 mg/mL of GL-PS, the rates of wound healing in mice were significantly higher and the wound healing time was significantly less than the control group (Plt;0.05 or Plt;0.01).
A certain concentration of GL-PS may promote wound healing via activation of the Wnt/β-catenin signaling pathway and up-regulation of TGF-β1, which might serve as a promising source of skin wound healing.
研究灵芝多糖(GL-PS)对人成纤维细胞及昆明雄性小鼠皮肤创伤愈合的影响,并探讨其可能的分子机制。
培养人皮肤原代成纤维细胞。分别用0、10、20、40、80和160μg/mL的GL-PS处理成纤维细胞,采用3-(4,5-二甲基-2-噻唑基)-2,5-二苯基-2-H-四氮唑溴盐(MTT)法检测细胞活力。采用Transwell法检测用0、10、20和40μg/mL的GL-PS处理的成纤维细胞的迁移能力。采用酶联免疫吸附测定法检测细胞上清液中I型前胶原C端肽(CICP)和转化生长因子-β1(TGF-β1)的分泌。采用蛋白质免疫印迹法检测β-连环蛋白的表达。此外,建立昆明小鼠全层皮肤切除创伤模型,分别外用10、20和40mg/mL的GL-PS进行处理。每天测量伤口大小,记录每组的完全愈合时间,并计算伤口收缩百分比。
与对照组相比,10、20和40μg/mL的GL-PS显著提高了成纤维细胞的活力,促进了成纤维细胞的迁移能力,并上调了成纤维细胞中CICP和TGF-β1的表达(P<0.05或P<0.01)。用20和40μg/mL的GL-PS处理的成纤维细胞中β-连环蛋白的表达显著高于对照组(P<0.01)。此外,外用10、20和40mg/mL的GL-PS后,小鼠的伤口愈合率显著提高,伤口愈合时间显著短于对照组(P<0.05或P<0.01)。
一定浓度的GL-PS可能通过激活Wnt/β-连环蛋白信号通路和上调TGF-β1促进伤口愈合,这可能是一种有前途的皮肤伤口愈合来源。