| 王智杰,王磊,郑益志.姜黄素对VEGF作用下的HaCat细胞的影响及KLF6/P21表达的调控研究[J].浙江中西医结合杂志,2019,29(5): |
| 姜黄素对VEGF作用下的HaCat细胞的影响及KLF6/P21表达的调控研究 |
| To Explore the Effect of Curcumin on HaCaT cells under the Influence ofVEGF and the Expression of KLF6/P21 |
| 投稿时间：2019-01-29 修订日期：2019-04-08 |
| DOI： |
| 中文关键词: 银屑病,姜黄素,HaCat细胞,VEGF,KLF6/P21 |
| 英文关键词:psoriasis, curcumin, HaCaT cell, VEGF, KLF6/P21 |
| 基金项目:1浙江省自然科学基金项目(LY14H110003)；2浙江省医药卫生科技计划项目(2013KYB185) |
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| 中文摘要: |
| 目的 通过观察姜黄素和VEGF对HaCat细胞增殖、凋亡及KLF6/P21蛋白表达的影响,探讨姜黄素治疗银屑病的可能机制,为姜黄素在银屑病治疗中进一步开发应用提供理论和实验基础。方法1.不同浓度的姜黄素(0,5,10,15,20,30,40μmol/L)作用于HaCat细胞,CCK8法检测其对细胞增殖的影响;20μmol/L姜黄素及25ng/ml VEGF分别及联合作用于HaCaT细胞,CCK8法检测20μmol/L姜黄素对25ng/L VEGF干预下HaCaT细胞的增殖作用。2.20μmol/L姜黄素及25ng/ml VEGF分别及联合作用于HaCat细胞,免疫蛋白印迹法检测KLF6/P21蛋白表达的改变。3.20μmol/L姜黄素及25ng/ml VEGF分别及联合作用于HaCat细胞,流式细胞仪检测其对HaCat细胞凋亡的影响。结果 1.姜黄素在0μmol/L-40μmol/L浓度范围内呈浓度依赖性抑制HaCat细胞增殖,20umol/L姜黄素明显抑制HaCat细胞增殖(P<0.01),在0μmol/L-40μmol/L范围内姜黄素浓度与HaCat细胞增殖呈线性相关(r=-0.92,P＜0.01)；与空白对照组相比,25ng/mL VEGF对HaCat细胞有明显的促增殖作用(P<0.05)。与25ng/L VEGF组相比20μmol/L姜黄素明显抑制25ng/L VEGF对HaCat细胞的促增殖作用(P<0.01)。2.25ng/mL VEGF提高HaCat细胞KLF6/P21蛋白的表达；20μmol/L姜黄素抑制HaCat细胞KLF6/P21蛋白的表达；20μmol/L姜黄素能降低25ng/mL VEGF对HaCat细胞KLF6/P21蛋白表达的增强作用。3.与空白对照组相比,20μmol/L姜黄素可促进HaCat细胞凋亡；与25ng/mL VEGF比较,20μmol/L姜黄素能明显抑制25ng/mL VEGF细胞的促增殖作用,促进细胞凋亡。结论 姜黄素可以抑制VEGF对HaCat细胞的促增殖作用,促进HaCat细胞凋亡,其机制可能与姜黄素可降低HaCat细胞KLF6/P21蛋白的表达有关。 |
| 英文摘要: |
| Objective To explore the possible mechanism of the curcumin treatment of psoriasis by observing the effects of curcumin and VEGF on HaCaT cell proliferation,apoptosis and KLF6/P21 protein expression.To provide an experimental basis for the further development and application of curcumin in the treatment of psoriasis.Methods 1.CCK8 assay was used to determine the proliferation of HaCaT cells incubated with different concentrations of curcumin(0,5,10,15,20,30,40μmol/L).The effects of 20μmol/L curcumin on the proliferation of HaCaT cells induced by 25ng/ml VEGF werealso tested with CCK8 assay.2. 20μmol/L and 25ng/ml VEGF respectively and combined with curcumin in HaCat cells, change the Western blot to detect the expression of KLF6/P21.3.Flow cytometry was used to detect the effects of curcumin and VEGF on HaCaT cell apoptosis.Results 1.The experimental result of CCK8 assay showed that in the range of 0-40μmol/L,curcumin decreased the proliferation of HaCaT cells in a dose-dependent manner,20μmol/L curcumin played a significant effect on the HaCaT cells(p<0.01), In the range of 0-40μmol/L,the concentration of curcumin was linearly correlated to the proliferation of HaCaT cells(r=-0.92,P＜0.01).Compared with the blank control group,20μmol/L curcumin could obviously inhibit the proliferation of HaCaT cells(P<0.01),20umol/L curcumin could obviously inhibit the promoting proliferation of 25 ng/ml VEGF in HaCaT cells(P<0.01).2.The results of Western-Blot showed that 25ng/mL VEGF promoted the expression of KLF6/P21 protein in HaCat cells;20μmol/L curcumin inhibiting expression of KLF6/P21 in HaCaT cell and decreased the effect of 25ng/ml VEGF on promoted expression of KLF6/P21 in HaCaT cell.3.The results of Flow cytometry showed that compared with the control group,20μmol/L curcumin can promote HaCaT cell apoptosis,compared with 25ng/ml VEGF group,20μmol/L curcumin could significantly inhibited the cell proliferation in duced by 25ng/ml VEGF.Conclusion Curcumin inhibit proliferation effect of VEGF on HaCaT cells and promote apoptosis in HaCaT cells,which mechanism may be relate to the inhibiting of the expression of KLF6/P21 protein in HaCaT cells. |
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