Diabetic foot ulcers(DFUs)represent a significant global healthcare burden due to their chronic nature and poor healing response.Gynura cusimbua,a medicinal plant with traditional wound-healing applications,was invest...Diabetic foot ulcers(DFUs)represent a significant global healthcare burden due to their chronic nature and poor healing response.Gynura cusimbua,a medicinal plant with traditional wound-healing applications,was investigated for its therapeutic potential in DFU management using an integrative approach involving phytochemical profiling,network pharmacology,molecular docking,MD simulation,and angiogenesis studies.OHRLC-MS analysis identified seven major phytochemicals in the ethyl acetate fraction,which were further screened for drug-likeness and target prediction.Network pharmacology analysis identified Protein Kinase C Alpha(PRKCA)as a key target associated with DFU pathology,particularly in modulating angiogenesis,inflammation,and oxidative stress.Molecular docking studies were performed to assess the binding affinity of four selected phytochemicals—Haplophytine,Hexahydrodifenidol,1-[(1-tert-Butyl-1H-tetrazol-5-yl)-(2,4-dimethoxyphenyl)-methyl]-4-cyclohexylpiperazine,and(3β,4α)-3-Hydroxycholest-5-en-4-yl acetate—against PRKCA,with Phorbol-12-myristate-13-acetate(PMA)serving as a reference activator.Among these,Haplophytine and 1-[(1-tert-Butyl-1H-tetrazol-5-yl)-(2,4-dimethoxyphenyl)-methyl]-4-cyclohexylpiperazine(6.8 kcal/mol)exhibited the strongest binding affinities,surpassing PMA.These two ligands were taken forward for MD simulations,which confirmed their superior structural stability,hydrogen bonding interactions,and lower solvent exposure making them the most promising PRKCA activators.Angiogenesis was evaluated using the chorioallantoic membrane assay,demonstrating a dose-dependent increase in secondary and tertiary blood vessel formation,with the highest angiogenic activity observed at 45μg/mL.Collectively,these findings suggest that Gynura cusimbua exerts a multi-targeted therapeutic effect on DFU through PRKCA activation and enhanced angiogenesis,highlighting its potential for phytotherapeutic applications in diabetic wound healing.展开更多
目的宫颈癌变过程中包括表观基因与调控基因的改变,以DNA甲基化为代表的表观遗传改变可导致宫颈癌的发生。本研究通过分析基因甲基化在早期宫颈腺癌组织中的表达差异性,寻找预测宫颈腺癌发生发展的分子标志物。方法收集新疆医科大学附...目的宫颈癌变过程中包括表观基因与调控基因的改变,以DNA甲基化为代表的表观遗传改变可导致宫颈癌的发生。本研究通过分析基因甲基化在早期宫颈腺癌组织中的表达差异性,寻找预测宫颈腺癌发生发展的分子标志物。方法收集新疆医科大学附属肿瘤医院2015-01-01-2017-06-30保存的新鲜初治原发宫颈腺癌组织15例和宫颈鳞状细胞癌组织15例,以及同期住院的正常宫颈组织15例,应用Illumina 850K甲基化芯片筛选宫颈腺癌组织的特异甲基化表达基因,亚硫酸盐测序法(bisulfite sequencing PCR,BSP)及实时定量RT-PCR(quantitative real time RT-PCR,Q-RT-PCR)方法对宫颈腺癌组织、宫颈鳞状细胞癌组织及正常宫颈组织进行验证。结果芯片筛选结果显示,性别决定区Y框蛋白1(sex determining region Y-box1,SOX1)、细胞周期蛋白D1(Cyclin D1,CCND1)和蛋白激酶Cα(protein kinase Cα,PRKCA)等基因是宫颈腺癌组织中基因调控网络的中心节点。BSP检测结果显示,SOX1基因甲基化程度在宫颈腺癌为(0.975±0.013)%,宫颈鳞癌为(0.871±0.073)%,正常宫颈组织为(0.416±0.256)%,差异有统计学意义,F=18.640,P<0.001。SOX1是宫颈腺癌特异甲基化表达基因。CCND1(F=0.890,P=0.436)和PRKCA(F=1.749,P=0.693)不是宫颈腺癌特异甲基化表达基因,差异均无统计学意义。Q-RT-PCR检测结果发现,SOX1(t=3.213,P=0.009)、PRKCA(t=-3.448,P=0.006)在宫颈腺癌组与宫颈鳞癌组比较差异均有统计学意义。SOX1(t=2.190,P=0.043)、PRKCA(t=-2.561,P=0.028)在宫颈腺癌组与正常宫颈组比较差异均有统计学意义,CCND1在宫颈腺癌组与正常宫颈组(t=3.937,P=0.003)、宫颈鳞癌组与正常宫颈组(t=4.100,P=0.002)差异均有统计学意义。结论抑癌基因SOX1是宫颈腺癌特异甲基化表达基因,有望成为宫颈腺癌诊断的特异标志物。CCND1、PRKCA基因不能证明是宫颈腺癌特异甲基化表达基因,考虑与BSP检测样本量少、检测位点少有关,有待于进一步研究。展开更多
文摘Diabetic foot ulcers(DFUs)represent a significant global healthcare burden due to their chronic nature and poor healing response.Gynura cusimbua,a medicinal plant with traditional wound-healing applications,was investigated for its therapeutic potential in DFU management using an integrative approach involving phytochemical profiling,network pharmacology,molecular docking,MD simulation,and angiogenesis studies.OHRLC-MS analysis identified seven major phytochemicals in the ethyl acetate fraction,which were further screened for drug-likeness and target prediction.Network pharmacology analysis identified Protein Kinase C Alpha(PRKCA)as a key target associated with DFU pathology,particularly in modulating angiogenesis,inflammation,and oxidative stress.Molecular docking studies were performed to assess the binding affinity of four selected phytochemicals—Haplophytine,Hexahydrodifenidol,1-[(1-tert-Butyl-1H-tetrazol-5-yl)-(2,4-dimethoxyphenyl)-methyl]-4-cyclohexylpiperazine,and(3β,4α)-3-Hydroxycholest-5-en-4-yl acetate—against PRKCA,with Phorbol-12-myristate-13-acetate(PMA)serving as a reference activator.Among these,Haplophytine and 1-[(1-tert-Butyl-1H-tetrazol-5-yl)-(2,4-dimethoxyphenyl)-methyl]-4-cyclohexylpiperazine(6.8 kcal/mol)exhibited the strongest binding affinities,surpassing PMA.These two ligands were taken forward for MD simulations,which confirmed their superior structural stability,hydrogen bonding interactions,and lower solvent exposure making them the most promising PRKCA activators.Angiogenesis was evaluated using the chorioallantoic membrane assay,demonstrating a dose-dependent increase in secondary and tertiary blood vessel formation,with the highest angiogenic activity observed at 45μg/mL.Collectively,these findings suggest that Gynura cusimbua exerts a multi-targeted therapeutic effect on DFU through PRKCA activation and enhanced angiogenesis,highlighting its potential for phytotherapeutic applications in diabetic wound healing.
文摘目的宫颈癌变过程中包括表观基因与调控基因的改变,以DNA甲基化为代表的表观遗传改变可导致宫颈癌的发生。本研究通过分析基因甲基化在早期宫颈腺癌组织中的表达差异性,寻找预测宫颈腺癌发生发展的分子标志物。方法收集新疆医科大学附属肿瘤医院2015-01-01-2017-06-30保存的新鲜初治原发宫颈腺癌组织15例和宫颈鳞状细胞癌组织15例,以及同期住院的正常宫颈组织15例,应用Illumina 850K甲基化芯片筛选宫颈腺癌组织的特异甲基化表达基因,亚硫酸盐测序法(bisulfite sequencing PCR,BSP)及实时定量RT-PCR(quantitative real time RT-PCR,Q-RT-PCR)方法对宫颈腺癌组织、宫颈鳞状细胞癌组织及正常宫颈组织进行验证。结果芯片筛选结果显示,性别决定区Y框蛋白1(sex determining region Y-box1,SOX1)、细胞周期蛋白D1(Cyclin D1,CCND1)和蛋白激酶Cα(protein kinase Cα,PRKCA)等基因是宫颈腺癌组织中基因调控网络的中心节点。BSP检测结果显示,SOX1基因甲基化程度在宫颈腺癌为(0.975±0.013)%,宫颈鳞癌为(0.871±0.073)%,正常宫颈组织为(0.416±0.256)%,差异有统计学意义,F=18.640,P<0.001。SOX1是宫颈腺癌特异甲基化表达基因。CCND1(F=0.890,P=0.436)和PRKCA(F=1.749,P=0.693)不是宫颈腺癌特异甲基化表达基因,差异均无统计学意义。Q-RT-PCR检测结果发现,SOX1(t=3.213,P=0.009)、PRKCA(t=-3.448,P=0.006)在宫颈腺癌组与宫颈鳞癌组比较差异均有统计学意义。SOX1(t=2.190,P=0.043)、PRKCA(t=-2.561,P=0.028)在宫颈腺癌组与正常宫颈组比较差异均有统计学意义,CCND1在宫颈腺癌组与正常宫颈组(t=3.937,P=0.003)、宫颈鳞癌组与正常宫颈组(t=4.100,P=0.002)差异均有统计学意义。结论抑癌基因SOX1是宫颈腺癌特异甲基化表达基因,有望成为宫颈腺癌诊断的特异标志物。CCND1、PRKCA基因不能证明是宫颈腺癌特异甲基化表达基因,考虑与BSP检测样本量少、检测位点少有关,有待于进一步研究。
