To investigate the mechanisms of apigenin(API)and proanthocyanidins(PC)in soothing sensitive skin(SS),a mast cell degranulation model was established by stimulating RBL-2H3 cells with the calcium ionophore A23187.Base...To investigate the mechanisms of apigenin(API)and proanthocyanidins(PC)in soothing sensitive skin(SS),a mast cell degranulation model was established by stimulating RBL-2H3 cells with the calcium ionophore A23187.Based on the combinatorial experiments,it was found that when API and PC were combined at the molar ratios of 4∶1 and 2∶1,they exhibited the antagonistic effects on histamine release(combination index CI>1);when they are combined at the molar ratios of 1∶1,1∶2 or 1∶4,they showed the synergistic effects on histamine release(CI<1).Among them,the combination of API and PC at a molar ratio of 1∶1 showed the better potent synergistic antihistamine release effect(CI=0.70).Histamine is a hallmark of the mast cell degranulation,consequently,the combination of API and PC at a molar ratio of 1∶1 yields the better efficiency in inhibiting the mast cell degranulation with the lowest IC_(50)value.Compared to the utilization of API or PC alone,the IC_(50)value was reduced by 11.150 and 6.503μmol/L,respectively.Compared to the positive control paeonol(PA),the treatment with the combination significantly reduced theβ-hex secretion,decreased the F-actin cytoskeleton rearrangement,and markedly suppressed the release of TNF-α,IL-4,and MCP-1.Further studies on the signaling pathways related to the mast cell degranulation indicated that the combination effectively inhibited the intracellular Ca^(2+)influx and significantly suppressed the phosphorylation of calmodulin-dependent protein kinase(CaMK)and phospholipase C/protein kinase C(PLC/PKC).In summary,the combination of API and PC at a molar ratio of 1∶1 exhibited the better synergistic antagonistic effect on the histamine release,inhibited the mast cell degranulation model activation by reducing Ca^(2+)influx and inhibiting the activation of Ca^(2+)/CaMK and PLC/PKC pathways,stabilized the cell membranes,regulated the inflammatory factor secretion,and exerted an effect in alleviating sensitive skin.展开更多
文摘To investigate the mechanisms of apigenin(API)and proanthocyanidins(PC)in soothing sensitive skin(SS),a mast cell degranulation model was established by stimulating RBL-2H3 cells with the calcium ionophore A23187.Based on the combinatorial experiments,it was found that when API and PC were combined at the molar ratios of 4∶1 and 2∶1,they exhibited the antagonistic effects on histamine release(combination index CI>1);when they are combined at the molar ratios of 1∶1,1∶2 or 1∶4,they showed the synergistic effects on histamine release(CI<1).Among them,the combination of API and PC at a molar ratio of 1∶1 showed the better potent synergistic antihistamine release effect(CI=0.70).Histamine is a hallmark of the mast cell degranulation,consequently,the combination of API and PC at a molar ratio of 1∶1 yields the better efficiency in inhibiting the mast cell degranulation with the lowest IC_(50)value.Compared to the utilization of API or PC alone,the IC_(50)value was reduced by 11.150 and 6.503μmol/L,respectively.Compared to the positive control paeonol(PA),the treatment with the combination significantly reduced theβ-hex secretion,decreased the F-actin cytoskeleton rearrangement,and markedly suppressed the release of TNF-α,IL-4,and MCP-1.Further studies on the signaling pathways related to the mast cell degranulation indicated that the combination effectively inhibited the intracellular Ca^(2+)influx and significantly suppressed the phosphorylation of calmodulin-dependent protein kinase(CaMK)and phospholipase C/protein kinase C(PLC/PKC).In summary,the combination of API and PC at a molar ratio of 1∶1 exhibited the better synergistic antagonistic effect on the histamine release,inhibited the mast cell degranulation model activation by reducing Ca^(2+)influx and inhibiting the activation of Ca^(2+)/CaMK and PLC/PKC pathways,stabilized the cell membranes,regulated the inflammatory factor secretion,and exerted an effect in alleviating sensitive skin.
文摘目的探讨黄酮类化合物芹菜素对大鼠实验性心肌缺血/再灌注(ischemia/reperfusion,I/R)时心肌细胞凋亡与Bcl-2、Bax、Caspase-3蛋白表达的影响,并分析心肌组织病理学损伤程度。方法采用结扎左冠状动脉前降支,心肌缺血45 min,再灌注2 h制作缺血/再灌注模型。将大鼠随机分为8组,即正常组(normal group,Normal)、假手术组(sham oper-ation group,Sham)、生理盐水缺血/再灌注组(saline ischemi-a-reperfusion group,NS)、溶剂对照组(solvent control group,Sol)、美托洛尔对照组(metoprolol control group,Meto)、芹菜素低、中、高剂量(1、2、4 mg.kg-1)用药组(apigenin low,medium and high dose treatment group,Api1,Api2,Api4)。再灌注2 h后迅速取出心脏,TUNEL法原位标记凋亡的心肌细胞;免疫组化法测Bcl-2、Bax和Caspase-3蛋白表达;做病理组织切片检查心肌损伤情况。结果芹菜素各剂量组心肌细胞凋亡率明显低于NS组(P<0.05),Api1,Api2,Api4能剂量依赖性地降低大鼠缺血/再灌注心肌细胞凋亡率;芹菜素各剂量组剂量依赖性地提高大鼠心肌缺血/再灌注的Bcl-2蛋白表达量(P<0.05)、降低大鼠心肌缺血/再灌注的Bax、Caspase-3蛋白表达量(P<0.05);芹菜素各剂量组与NS组比较,心肌组织损伤的病理学变化明显减轻(P<0.05)。结论芹菜素对缺血/再灌注心肌的保护效应可能与其抑制缺血/再灌注心肌细胞凋亡有关;芹菜素抗心肌凋亡作用的机制可能与其上调Bcl-2蛋白表达和下调Bax、Caspase-3蛋白表达有关;芹菜素能明显减轻心肌组织损伤。
