目的探究刺梨根治疗寒邪客胃型胃溃疡的作用及机制。方法将SPF级雄性Wistar大鼠按照随机数表法分为三组,分别为A组(对照组)、B组(模型组)以及C组(治疗组),每组3只。采用冰乙酸浆膜下注射法制备大鼠胃溃疡模型,建模完成后开始给药,C组(...目的探究刺梨根治疗寒邪客胃型胃溃疡的作用及机制。方法将SPF级雄性Wistar大鼠按照随机数表法分为三组,分别为A组(对照组)、B组(模型组)以及C组(治疗组),每组3只。采用冰乙酸浆膜下注射法制备大鼠胃溃疡模型,建模完成后开始给药,C组(治疗组)给予每天2 m L/100 g刺梨根煎液灌胃处理,1次/天,连续灌胃14 d。观察每组大鼠的整体精神状态、活动情况、毛发光泽和粪便性状等。给药结束后1天,处死动物,取材外周血以及胃黏膜组织。观察新鲜胃黏膜样本,计算胃黏膜溃疡指数及溃疡抑制率,并通过石蜡包埋后,HE染色观察胃黏膜病理组织变化。用ELISA法测定血清表皮生长因子(EGF)、白细胞介素-6(IL-6)水平。结果治疗组大鼠一般状态优于模型组。与对照组相比,模型组胃黏膜层结构相对完整,腺体局部排列紊乱,出现不同程度的黏膜水肿糜烂,上皮细胞疏松脱落,可见大量炎性细胞浸润;IL-6水平升高,EGF水平降低。加入刺梨根煎液干预后,大鼠胃黏膜层、固有层、肌层结构相对完整,固有腺体排列较整齐,未见明显的炎性细胞浸润;溃疡指数降低,IL-6水平降低,EGF水平升高,差异具有统计学意义(P<0.05)。结论刺梨根可以通过调控EGF、IL-6水平,起到调节攻击因子与保护因子的平衡来保护胃黏膜的作用。展开更多
Aim To study the proliferative effeet of hydroxysaftlor yellow A （HSYA） on cultured canine aortic endothelial cell （VEC） in normoxic （21% O2 ） or hypoxic （10% O2 ） culture and the underlying mechanism. Methods...Aim To study the proliferative effeet of hydroxysaftlor yellow A （HSYA） on cultured canine aortic endothelial cell （VEC） in normoxic （21% O2 ） or hypoxic （10% O2 ） culture and the underlying mechanism. Methods The endothelial cells were scratched from trypsined canine aorta endothelium. HSYA was added to the cells at final concentrations of 1 × 10^-3, 1 × 10^-4 and 1 × 10^-5 mol· L^-1, respectively. VEGF （2.6 × 10^-7 mol· L^-1 ）-treated cells were used as the positive control. The proliferative effect of HSYA on VEC was determined at 48, 72, 96, and 120 h in normoxic culture by MTI＂ assay. Similarly, the proliferation of VEC was determined at 12, 24, 48, and 72 h in hypoxic culture by MTF assay. The effects of HSYA on VEC proliferation and VEGF secretion were investigated by MTr and ELISA assays at the presence of the antibodies to VEGF and VEGF receptors. Results Pretreatment with HSYA at concentrations of 1 × 10^-3 and 1 × 10^-4 mol· L^-1 enhanced VEC proliferation in normoxic culture. The most significant enhancing effect of HSYA on VEC proliferation was achieved at 24, 48, and 72 h in hypoxic culture in concentration-dependent and time-dependent manner. HSYA at 1 × 10^-3 mol·L^-1 showed a potency similar to VEGF at 2.6 × 10^-7 mol·L^-1 . Pretreatment with the antibodies of Flt-1, KDR or VEGF blocked the proliferative effect of HSYA with similar potencies. Antibodies of Fit-1 or VEGF antagonized the promoting effect of HSYA on VEGF secretion. Conclusion HSYA promotes VEC proliferation either in normoxic or hypoxic culture, especially in the latter condition. This effect of HSYA is at least partly mediated by VEGF and VEGF receptor.展开更多
目的观察土茯苓提取物落新妇苷对永生化人角质形成细胞Ha Ca T增殖、凋亡的影响,并探讨其机制。方法收集对数生长期Ha Ca T细胞,观察组分别加入不同浓度落新妇苷、空白对照组加DMEM培养液,培养24 h后,采用MTT法测算细胞增殖抑制率,流式...目的观察土茯苓提取物落新妇苷对永生化人角质形成细胞Ha Ca T增殖、凋亡的影响,并探讨其机制。方法收集对数生长期Ha Ca T细胞,观察组分别加入不同浓度落新妇苷、空白对照组加DMEM培养液,培养24 h后,采用MTT法测算细胞增殖抑制率,流式细胞仪检测细胞早期凋亡率;收集对数生长期Ha Ca T细胞,观察组分别加入不同浓度落新妇苷+20 ng/m L INF-γ、模型对照组加20 ng/m L INF-γ、空白对照组加DMEM培养液培养24 h,采用实时荧光定量PCR法检测细胞中的核转录因子κB(NF-κB)p65、骨桥蛋白(OPN)、血管内皮生长因子(VEGF)mRNA。结果落新妇苷在25、50、100、200、400μg/m L时,Ha Ca T细胞增殖抑制率分别为3.06%、26.53%、40.82%、60.20%及61.22%。落新妇苷在50、100、200μg/m L时,观察组Ha Ca T细胞早期凋亡率分别为3.26%、8.24%和15.38%;与空白对照组早期凋亡率(1.33%)比较,P均<0.05。与空白对照组比较,模型对照组NF-κB p65、OPN、VEGF mRNA表达增加(P均<0.01);与模型对照组比较,观察组随落新妇苷浓度增加,NF-κB p65、OPN、VEGF mRNA表达逐渐降低(P均<0.05)。结论土茯苓提取物落新妇苷能抑制Ha Ca T细胞增殖、诱导细胞凋亡,其机制可能与制抑细胞NF-κB p65、OPN、VEGF mRNA表达有关。展开更多
文摘目的探究刺梨根治疗寒邪客胃型胃溃疡的作用及机制。方法将SPF级雄性Wistar大鼠按照随机数表法分为三组,分别为A组(对照组)、B组(模型组)以及C组(治疗组),每组3只。采用冰乙酸浆膜下注射法制备大鼠胃溃疡模型,建模完成后开始给药,C组(治疗组)给予每天2 m L/100 g刺梨根煎液灌胃处理,1次/天,连续灌胃14 d。观察每组大鼠的整体精神状态、活动情况、毛发光泽和粪便性状等。给药结束后1天,处死动物,取材外周血以及胃黏膜组织。观察新鲜胃黏膜样本,计算胃黏膜溃疡指数及溃疡抑制率,并通过石蜡包埋后,HE染色观察胃黏膜病理组织变化。用ELISA法测定血清表皮生长因子(EGF)、白细胞介素-6(IL-6)水平。结果治疗组大鼠一般状态优于模型组。与对照组相比,模型组胃黏膜层结构相对完整,腺体局部排列紊乱,出现不同程度的黏膜水肿糜烂,上皮细胞疏松脱落,可见大量炎性细胞浸润;IL-6水平升高,EGF水平降低。加入刺梨根煎液干预后,大鼠胃黏膜层、固有层、肌层结构相对完整,固有腺体排列较整齐,未见明显的炎性细胞浸润;溃疡指数降低,IL-6水平降低,EGF水平升高,差异具有统计学意义(P<0.05)。结论刺梨根可以通过调控EGF、IL-6水平,起到调节攻击因子与保护因子的平衡来保护胃黏膜的作用。
文摘Aim To study the proliferative effeet of hydroxysaftlor yellow A （HSYA） on cultured canine aortic endothelial cell （VEC） in normoxic （21% O2 ） or hypoxic （10% O2 ） culture and the underlying mechanism. Methods The endothelial cells were scratched from trypsined canine aorta endothelium. HSYA was added to the cells at final concentrations of 1 × 10^-3, 1 × 10^-4 and 1 × 10^-5 mol· L^-1, respectively. VEGF （2.6 × 10^-7 mol· L^-1 ）-treated cells were used as the positive control. The proliferative effect of HSYA on VEC was determined at 48, 72, 96, and 120 h in normoxic culture by MTI＂ assay. Similarly, the proliferation of VEC was determined at 12, 24, 48, and 72 h in hypoxic culture by MTF assay. The effects of HSYA on VEC proliferation and VEGF secretion were investigated by MTr and ELISA assays at the presence of the antibodies to VEGF and VEGF receptors. Results Pretreatment with HSYA at concentrations of 1 × 10^-3 and 1 × 10^-4 mol· L^-1 enhanced VEC proliferation in normoxic culture. The most significant enhancing effect of HSYA on VEC proliferation was achieved at 24, 48, and 72 h in hypoxic culture in concentration-dependent and time-dependent manner. HSYA at 1 × 10^-3 mol·L^-1 showed a potency similar to VEGF at 2.6 × 10^-7 mol·L^-1 . Pretreatment with the antibodies of Flt-1, KDR or VEGF blocked the proliferative effect of HSYA with similar potencies. Antibodies of Fit-1 or VEGF antagonized the promoting effect of HSYA on VEGF secretion. Conclusion HSYA promotes VEC proliferation either in normoxic or hypoxic culture, especially in the latter condition. This effect of HSYA is at least partly mediated by VEGF and VEGF receptor.
文摘目的观察土茯苓提取物落新妇苷对永生化人角质形成细胞Ha Ca T增殖、凋亡的影响,并探讨其机制。方法收集对数生长期Ha Ca T细胞,观察组分别加入不同浓度落新妇苷、空白对照组加DMEM培养液,培养24 h后,采用MTT法测算细胞增殖抑制率,流式细胞仪检测细胞早期凋亡率;收集对数生长期Ha Ca T细胞,观察组分别加入不同浓度落新妇苷+20 ng/m L INF-γ、模型对照组加20 ng/m L INF-γ、空白对照组加DMEM培养液培养24 h,采用实时荧光定量PCR法检测细胞中的核转录因子κB(NF-κB)p65、骨桥蛋白(OPN)、血管内皮生长因子(VEGF)mRNA。结果落新妇苷在25、50、100、200、400μg/m L时,Ha Ca T细胞增殖抑制率分别为3.06%、26.53%、40.82%、60.20%及61.22%。落新妇苷在50、100、200μg/m L时,观察组Ha Ca T细胞早期凋亡率分别为3.26%、8.24%和15.38%;与空白对照组早期凋亡率(1.33%)比较,P均<0.05。与空白对照组比较,模型对照组NF-κB p65、OPN、VEGF mRNA表达增加(P均<0.01);与模型对照组比较,观察组随落新妇苷浓度增加,NF-κB p65、OPN、VEGF mRNA表达逐渐降低(P均<0.05)。结论土茯苓提取物落新妇苷能抑制Ha Ca T细胞增殖、诱导细胞凋亡,其机制可能与制抑细胞NF-κB p65、OPN、VEGF mRNA表达有关。
