摘要
目的探讨牡丹皮功效部位含药血清对过氧化氢(H_(2)O_(2))诱导损伤的人脐静脉内皮细胞(HUVEC)的保护作用和机制。方法水蒸气蒸馏法和乙醇提取上大孔吸附树脂柱精制法提取丹皮酚和丹皮总苷。制备含药血清。采用H_(2)O_(2)诱导损伤HUVEC,与丹皮酚、丹皮总苷含药血清共培养后,MTT法检测细胞存活率;微板法检测细胞上清液中乳酸脱氢酶(LDH)、内皮素-1(ET-1)和一氧化氮(NO)的含量;流式细胞术检测细胞凋亡率;Western Blot法测定Bax、Bcl-2蛋白表达水平。结果与空白组比较,模型组细胞的LDH、ET-1含量均有显著提高(P<0.01),而NO含量显著降低(P<0.01);与模型组比较,丹皮总苷、丹皮酚、联合给药组的细胞存活率明显提高(P<0.05或P<0.01)且细胞凋亡减少;丹皮总苷、丹皮酚及联合给药组的LDH含量显著降低(P<0.01或P<0.05);丹皮总苷、联合给药组NO含量显著提高(P<0.01);丹皮总苷、联合给药组ET-1含量显著降低(P<0.05或P<0.01)。丹皮总苷、丹皮酚、联合给药组均能下调Bax蛋白、上调Bcl-2蛋白表达水平(P<0.05或P<0.01)。结论丹皮酚和丹皮总苷配伍对H_(2)O_(2)诱导损伤的HUVEC有增效作用,其机制可能与调节Bcl-2/Bax比值抑制细胞凋亡有关。
Objective To investigate the effect and mechanism of active fractions from Mudanpi (Cortex Moutan) on apoptosis of human umbilical vein endothelial cells (HUVECs) induced by hydrogen peroxide (H_(2)O_(2)). Methods Paeonol and total glucosides were extracted from Mudanpi (Cortex Moutan) by steam distillation and ethanol extraction. The serum containing correspondent drugs were prepared. The model of HUVECs injury induced by H_(2)O_(2) was established, and the serum containing different active fractions from Mudanpi (Cortex Moutan) was used to intervene. The cell survival rate was detected by MTT method. The contents of lactate dehydrogenase (LDH), endothelin-1 (ET-1) and nitric oxide (NO) in cell supernatant were detected by microplate method. The apoptotic rate was detected by flow cytometry. The expression levels of Bax and Bcl-2 protein were determined by Western Blot. Results Compared with those of the blank group, the contents of LDH and ET-1 in the model group were significantly increased (P<0.01), while the content of NO was significantly decreased (P<0.01). Compared with those of the model group, the cell survival rate was significantly increased and the apoptosis was decreased in the total glucosides, paeonol and combination groups (P<0.05 or P<0.01), while the contents of LDH in the total glucosides, paeonol and the combination groups were significantly lower than those in the model group (P<0.01 or P<0.05). The content of NO in total glucosides and combination groups increased significantly (P<0.01), while the content of ET-1 in total glucosides and combination groups decreased significantly (P<0.05 or P<0.01). Total glucosides, paeonol and combination group could down-regulate the expression of Bax protein and up-regulate the expression of Bcl-2 protein (P<0.05 or P<0.01). Conclusion The combination of paeonol and total glucosides of Mudanpi (Cortex Moutan) has a synergistic effect on H_(2)O_(2)-induced HUVEC injury, and its mechanism may be related to regulating the ratio of Bcl-2/Bax to inhibit cell apoptosis.
作者
吴冰帆
张留记
李开言
王建霞
屠万倩
WU Bingfan;ZHANG Liuji;LI Kaiyan;WANG Jianxia;TU Wanqian(Henan University of Traditional Chinese Medicine,Zhengzhou 450008,Henan,China;Henan Acudemy of Traditional Chinese Medicine,Zhengzhou 450004,Henan,China)
出处
《中华中医药学刊》
CAS
北大核心
2021年第4期209-212,共4页
Chinese Archives of Traditional Chinese Medicine
基金
河南省科技创新人才计划(144200510019)
河南省中医药科学研究专项(2016ZY1003)。
