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目的:探讨四君子加川牛膝方促进大鼠下肢难愈性创面愈合可能的作用机制,以及引经药川牛膝的作用。方法:建立SD大鼠右下肢难愈性创面模型,随机分为生理盐水组、四君子加川牛膝方组、四君子汤组、川牛膝方组,各组分别给予生理盐水及相应中药灌胃处理。在创面愈合过程中(造模后的第7天、第14天),取创面新生肉芽组织及血液,检测血清碱性成纤维生长因子(b FGF)、转化生长因子-β1(TGF-β1)水平的变化,并应用流式细胞仪检测成纤维细胞的S期细胞比例。结果:(1)造模后第7天,四君子汤组及四君子加川牛膝方组TGF-β1、b FGF浓度明显高于生理盐水组(P<0.05),而川牛膝方组与生理盐水组无明显变化(P>0.05),其中四君子汤组与四君子加川牛膝方组间比较无明显差异(P>0.05);造模后第14天,各组间TGF-β1、b FGF浓度比较均无明显差异(P>0.05)。(2)造模后第7天,各药物组成纤维细胞G0/G1期细胞比例的表达均明显低于生理盐水组(P<0.01或P<0.05)、S期细胞比例的表达均明显高于生理盐水组(P<0.01),而各组间G2/M期细胞比例的表达均无显著性差异(P>0.05),其中四君子加川牛膝方组G0/G1期细胞比例明显低于四君子汤组(P<0.05),S期细胞比例明显高于四君子汤组(P<0.05);在造模后第14天,各组间G0/G1期及G2/M细胞比例的表达均无显著性差异(P>0.05),而四君子加川牛膝方组S期细胞比例明显高于四君子汤组与川牛膝方组(P<0.01)。结论:四君子加川牛膝方可上调TGF-β1、bFGF的合成和分泌,加速成纤维细胞由G0/G1期进入S期,促进成纤维细胞的增殖,这可能为其促进肉芽组织增生及创面愈合的作用机制,其中引经药川牛膝有提高成纤维细胞S期细胞比例的协同作用。
Objective: To investigate the possible mechanism of Sijunan Jiachuan Achyranthes root to promote the healing of lower extremity refractory wounds, Methods: The right lower extremity refractory wound model of SD rats was established and randomly divided into normal saline group, Sijijieji plus Achyranthes bidentata group, Sijunzi decoction group and Chuan Aizhu decoction group. The rats in each group were given normal saline and the corresponding traditional Chinese medicine . During wound healing (7 days and 14 days after modeling), granulation tissue and blood were collected from fresh wounds and serum levels of bFGF, TGF-β1, The level of S phase cells in fibroblasts was detected by flow cytometry. Results: (1) The concentration of TGF-β1 and b FGF in the Sijunzi Decoction group and the Sijijiazhuang Achyranthes Root group were significantly higher than those in the saline group on the 7th day (P <0.05) (P> 0.05). There was no significant difference between Sijunzi Decoction group and Sijijiakachuan Achyranthes bidentata group (P> 0.05). On the 14th day after model establishment, the levels of TGF-β1 and b FGF in each group were compared No significant difference (P> 0.05). (2) On the 7th day after the model was established, the percentage of cells in G0 / G1 phase in each drug group was significantly lower than that in the saline group (P <0.01 or P <0.05), and the percentage of S phase cells was significantly higher than that in the saline group (P <0.01). There was no significant difference in the proportion of cells in G2 / M phase among all groups (P> 0.05). The proportion of cells in G0 / G1 phase in Sijunang plus Achyranthes bidentata group was significantly lower than that in Sijunzi decoction (P <0.05). The proportion of cells in S phase was significantly higher than those in Sijunzi Decoction group (P <0.05). On the 14th day after model establishment, the expression of G0 / G1 phase and G2 / M cells in each group had no significant difference (P> 0.05). However, the proportion of cells in the S phase in the Aijian Kaguan Achyranthes root group was significantly higher than that in the Sijunzi decoction group and the Chuan Aogen group (P <0.01). Conclusion: Sijunzi plus Achyranthes bidentata can up-regulate the synthesis and secretion of TGF-β1, bFGF and accelerate the progression of fibroblasts from G0 / G1 phase to S phase and promote the proliferation of fibroblasts, which may be the promotion of granulation tissue proliferation and wound healing The mechanism of action, which cited Achyranthes bidentata enhance the synergistic effect of fibroblast S phase cell ratio.