Effect of astragalus injection on fibroblasts of human hypertrophic scar

WANG Qixin; WANG Xiaona; LIU Ruidi; GUO Bingyu; YANG Dan; XIE Bing; FU Zhiqiang

doi:10.3969/j.issn.1673-7040.2025.11.010

Abstract：Objective The effect of Astragalus injection on the growth of human hypertrophic scar fibroblasts(HSFB)was ex-plored.Methods From January to March 2024,sixteen patients who visited the Second Affiliated Hospital of Shenyang Medical College were selected.With the patients' consent,hypertrophic scar(HS)tissues removed during surgery were collected,and hypertrophic scar fi-broblasts(HSFB)were extracted from them for cell experiments.Eleven groups were observed(blank group,control group,Astragalus in-jection 5 mg/ml group,20 mg/ml group,40 mg/ml group,60 mg/ml group,80 mg/ml group,100 mg/ml group,120 mg/ml group,140 mg/ml group,160 mg/ml group),and they were cultured in vitro for 12,24,36,48 h respectively.The effect of different concentrations of Astra-galus injection on the growth ability of HSFB was observed by CCK8 assay,the inhibition rate was calculated,the IC50 curve was drawn,and the cytotoxicity and the relationship between the inhibition rate and the drug concentration and time were detected to select the appro-priate drug concentration for subsequent experiments.The effect of different concentrations of Astragalus injection on the migration ability of fibroblasts was observed by Transwell assay and scratch assay.The effect of different concentrations of Astragalus injection on the apop-tosis of HSFB was observed by Annexin Ⅴ-Propidium iodide(AV-PI)technology.The effect of different concentrations of Astragalus injec-tion on the expression of type Ⅰ collagen(CollagenⅠ),cell cycle protein(Cyclin D1)and apoptosis regulatory protein(Bel-2)in HSFB was detected by Western blot assay.Results The IC50 curve indicated that the inhibition rate of Astragalus injection on HSFB was time-and concentration-dependent,and it had a non-toxic inhibitory effect on cells.Five concentrations were selected for data analysis.The CCK8 assay results showed that after Astragalus injection intervened HSFB for 12,24,36,48 h,the 20 mg/ml,40 mg/ml,60 mg/ml and 80mg/ml drug concentration groups inhibited the proliferation of HSFB to varying degrees compared with the control group(P＜0.05).There was no significant difference in the OD value of HSFB between the 5 mg/ml group and the control group(P＞0.05);there was no significant differ-ence in the OD value of HSFB between the 80 mg/ml group and the 60 mg/ml group(P＞0.05).Subsequently,20 mg/ml,40 mg/ml and 60 mg/ml drug concentrations were used for experiments.The Transwell assay and scratch assay results showed that Astragalus injection inhibited the proliferation and migration of HSFB to varying degrees,and the inhibitory effect increased with the increase of Astragalus injection concen-tration(P＜0.05).The AV-PI assay results showed that Astragalus injection increased the apoptosis rate of HSFB to varying degrees(P＜0.05).The Western blot assay results showed that Astragalus injection down-regulated the expression of CollagenⅠ,Cyclin D1 and Bcl-2,and the down-regulation effect increased with the increase of Astragalus injection concentration(P＜0.05).Conclusion Astragalus injection can inhibit the proliferation and migration of HSFB and induce their apoptosis by down-regulating the expression of CollagenⅠ,Cyclin D1 and Bcl-2,and the effect is concentration-dependent.