The Function And Mechanism Of TGF-β1/mTOR Pathway In Polygonum Cuspidatum Resveratrol Inhibiting Proliferation Of Keloid Fibroblasts

Keloid is the pathological product of wound repair.Polygonum cuspidatum resveratrol(Res) is a kind of Traditional Chinese medicine (TCM)monomer. Res can induce keloid fibroblast apoptosis, which has certain curative effect in keloid. But the specific mechanism of the effect is not clear. A variety of genes and a great many of pathways regulate the formation of keloid.TGFβ1 is the most critical cytokines in keloid formation,which can affect multiple signaling pathways. Current research shows that mTOR play an important role in TGFβ1 signal transduction. mTOR pathway can not only affect protein synthesis,but also regulate the proliferation and differentiation of cells. mTOR signaling pathway mediates a variety of tumor diseases.The growth characteristics of keloid is similar to tumor, so keloid belongs to benign solid tumor. So in the process of keloid fibroblasts proliferation the mTOR pathway mediated by TGFβ1 may play a vital role. This subject research the expression of TGFβ1/mTOR pathway in keloid,and the impact of the Res in treating keloid and the effective targets.The purposeThis experiment will research the expression of TGF-β1、PI3K、AKT、mTOR which are the key molecules of TGF-β1/mTOR pathway in keloid、confirm the pathway expresses abnormally in keloid tissue、prove that Res can influence the TGF-β1/mTOR pathway in treating keloids、、clear targets of Res in treating keloids.The methodsThis experiment will study keloids and normal skin fibroblasts In vitro culture.There are three groups.Normal skin group (normal group) study on human normal skin fibroblasts;Keloid group research keloid fibroblasts;Resveratrol group(Res group)study keloid fibroblasts which was impacted by Resveratrol.(1) This experimental observe the distribution of the key moleculesTGF-β1、PI3K、AKT、mTOR in keloid fibroblasts by immunofluorescence technique. (2)This experimental detects each group the expression of TGF-β1 mRNA、PI3K mRNA、Akt mRNA、mTOR mRNA in TGF-β1/mTOR pathway by RT-PCR technique. (3)This experimental surveys the protein expression of every group TGF-β1、PI3K、AKT、mTOR in TGF-β1/mTOR pathway by Western blot technique.The resultsThe immunofluorescence results show that the key molecules TGF-β1、PI3K、Akt、mTOR express in the keloid fibroblasts cell nucleus and cytoplasm. According to the results of the RT-PCT,it shows that the expression of TGF-β1 mRNA、PI3K mRNA、Akt mRNA、mTOR mRNA in keloid group were higher than the quantity of TGF-β1 mRNA、P13K mRNA、Akt mTOR mRNA in normal group (P<0.05). It also shows that the expression of TGF-β1 mRNA、PI3K mRNA、Akt mRNA、mTOR mRNA in Res group were lower than that of keloid group(P<0.05). The results of Western blot showes that the protein expression of TGF-β1、 PI3K、Akt、mTOR in keloid group were higher than that in normal group(P<0.05).The protein expression of TGF-β1、PI3K、mTOR in Res group are lower than the resules of keloid group(P< 0.05),but the expression of Akt protein has no statistical difference between the Res group and keloid group(P>0.05),although the res group quantitative value is lower than that of keloid group.The conclusionsThe results shows that the TGF-β1/mTOR pathway is abnormal activity in keloid,while TGF-β1、PI3K、Akt and mTOR are the key molecules for their high expression. Res has inhibition impact on TGF-β1、PI3K、mTOR,which are the key molecules of TGF-β1/mTOR pathway in keloid. This inhibition may be the mechanism of action of Res in the treatment of keloid. This experimental study provide theoretical basis for the Res treatment of keloid.