The Study Of The Effect Of Ginsenoside Rg3 On Pathological Scars And The Relevant Mechanism Research

Objective The present study mainly focused on the effect of ginsenoside Rg3 with different concentrations on the pathological scars.On the basis of the results of previous animal experiments,we further defined the inhibitory effect of Rg3 on scars.Meanwhile,the comparative analysis of the genetic and molecular levels of several cytokines such as collagen I,collagen III,fibronectin,?-SMA,VEGF,etc were conducted to explore the role and function mechanism of TGF-?/Smad and Erk signaling pathways in scar formation,which will provide new mentality and strategy for the treatment of pathological scars.Materials and methods After excised from the patients,the scar tissues were digested by collagenase and primarily cultured.Then the primary cells were passaged and we chose the first to third generation to perform experiments.In every assay,KFs were divided into three groups(one control group and two experimental groups)and cultured in medium with or without different Rg3 concentrations(50 or 100 ?g/ml).We conducted the following experiments: microscopic observation,CCK-8 assay,flow cytometric(FCM)analysis,scratch wound assay,cell invasion assay,RNA isolation and quantitative PCR(Q-PCR),immunofluorescence staining,western blot,enzyme linked immunosorbent assay(ELISA)and keloid explant culture,thus analyzing the effect of Rg3 on pathological scar fibroblasts and exploring the mechanism.Results: 1.Rg3 suppressed scar fibroblasts proliferation in vitro in concentration-dependent manner.The expression of the proliferation marker Ki-67 in Rg3-treated group was prominently decreased.2.High concentration(100 ?g/ml)of Rg3 induced scar fibroblasts apoptosis.However,there was no obvious difference between the control group and 50 ?g/ml-Rg3-treated group.3.Rg3 inhibited the scar fibroblasts migration.4.Ginsenoside Rg3 suppressed the keloid fibroblasts invasion and the inhibitory effect was functioned in concentration-dependent manner.5.Rg3 reduced collagen production and ECM accumulation,leading to the down-regulation of the expression levels of pro-fibrogenic genes and proteins(Type I Collagen,Type III Collagen,Fibronectin,?-SMA,etc)and the up-regulation of anti-fibrosis genes levels(IFN-?,TGF-?3).6.The level of TGF-?1 and the phosphorylation levels of smad2 and smad3 were induced by Rg3 while the level of smad7 increased,which indicated that the TGF-?/Smad signaling pathway was involved in the process of inhibitory action of Rg3 on pathological scars.7.The vascularization of keloid tissue was obviously suppressed by Rg3.The expression level of VEGF and the phosphorylation level of ERK1/2 were induced,indicating that the ERK signaling pathway was involved in the process of inhibitory action of Rg3 on keloid.Conclusions: Ginsenoside Rg3 inhibited scar fibroblasts proliferation,invasion,angiogenesis and collagen accumulation.The TGF-??Smad and ERK1/2 signaling pathways were involved in these biological processes.Our experimental results showed that Rg3 may serve as a potential drug which can inhibit scar formation and growth in the clinical practice.This study will provide new mentality and strategy for the treatment of pathological scars.