Protective effects of fractions from Pseudostellaria heterophylla against cobalt chloride-induced hypoxic injury in H9c2 cell
Protective effects of fractions from Pseudostellaria heterophylla against cobalt chloride-induced hypoxic injury in H9c2 cell
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太子参提取物对氯化钴诱导的 H9c2 细胞缺氧损伤的保护作用
DOI：
10.1016/j.jep.2013.03.053
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发表时间：
2013-05-20
影响因子：
5.4
通讯作者：
Wang, Yong-Lin
中科院分区：
文献类型：
--
作者：
Wang, Zhen;Liao, Shang-Gao;Wang, Yong-Lin
民族药理学相关性：太子参除作为治疗脾虚、厌食、乏力的知名滋补中草药外，还被用于治疗心悸的中药。研究目的：探讨太子参提取物对缺氧心肌细胞的保护作用，观察到的保护作用是由哪些结构类型的化合物引起的，可能的作用机制是什么。材料与方法：用70%水乙醇和水先后提取太子参根，得到70%乙醇提取物和水提取物。后者首先用80%乙醇沉淀，然后用Sevag法去除蛋白质，得到富含多糖（PHP）的馏分。前者通过柱层析分离成富含小分子糖和氨基酸（PHSSAC）、皂苷（PHS）、环肽（PHCP）和皂苷元（PHSG）的组分。采用紫外光谱法或化学方法对五种组分进行确证。通过测定预处理后心肌细胞H9c2在cocl2诱导缺氧后的活力和乳酸脱氢酶（LDH）渗漏量，评价各组分的心脏保护作用。通过Annexin V/PI流式细胞术分析细胞死亡抑制的性质及其对丙二醛（MDA）、超氧化物歧化酶（SOD）和细胞内活性氧（ROS）水平的影响，研究其作用机制。结果：PHS和PHP组分对cocl2诱导的缺氧损伤的减弱作用高于或与阳性对照n -乙酰- l-半胱氨酸（NAC）相当。800 μ g/mL的PHS或10 mg/mL的PHP对细胞进行预处理后，MDA水平明显降低，细胞内ROS减少，SOD活性升高，LDH漏出量降低至接近或优于326 μ g/mL NAC的水平。两组细胞凋亡均减少。结论：综上所述，太子参治疗心悸的传统应用可能与太子参中含有心脏保护因子有关。PHP和PHS是其具有心脏保护作用的两个活性组分。其机制可能包括通过防止氧化应激增加来保护细胞膜免受缺氧损伤和细胞免受氧化损伤。通过抑制细胞凋亡来保护细胞也可能参与其中。2013爱思唯尔爱尔兰有限公司版权所有。
Ethnopharmacological relevance: Except for as a well-known tonic Chinese herbal medicine for the treatment of splenic asthenia, anorexia, lassitude and weakness, the roots of Pseudostellaria heterophylla was also used in Chinese medicines for the treatment of palpitation.Aim of the study: The study was designed to determine whether fractions from Pseudostellaria heterophylla could provide cardioprotection on hypoxic cardiomyocytes, what structural types of compounds were responsible for the observed effects, and which is the possible mechanism of action.Materials and Methods: The roots of Pseudostellaria heterophylla were extracted successively with 70% aqueous ethanol and water to give a 70% ethanol extract and a water extract. The latter was first precipitated by 80% ethanol and then protein-removed by the Sevag method to give a fraction enriched in polysaccarides (PHP). The former was separated by column chromatography into a fraction enriched in small-molecule sugars and amino acids (PHSSAC), saponins (PHS), cyclopeptides (PHCP), and sapogenins (PHSG). UV spectral or chemical methods were used to confirm the five fractions. The cardioprotective effects of the fractions were evaluated by measuring the viability and the leakage of lactate dehydrogenase (LDH) of the fraction-pretreated cardiomyocyte H9c2 after exposure to CoCl2-induced hypoxia. The mechanism of action was studied by investigating the nature of cell death inhibition (by Annexin V/PI flow cytometric analysis) and their effects on the levels of malonaldehyde (MDA), superoxide dismutase (SOD) and intracellular reactive oxygen species (ROS).Results: Fractions PHS and PHP could attenuate CoCl2-induced hypoxic damage to an extent higher than or comparable to the effect of the positive control N-acetyl-L-cysteine (NAC). Pretreatment of the cells with 800 mu g/mL of PHS or 10 mg/mL of PHP markedly decreased the level of MDA, reduced intracellular ROS, increased the activity of SOD, and reduced leakage of LDH to the levels close to or better than that with 326 mu g/mL of NAC. Reduction of apoptosis was also observed for both fractions.Conclusions: The overall results suggested that the traditional use of this plant for the treatment of palpitation may be attributed to the presence of cardioprotective agents in Pseudostellaria heterophylla. PHP and PHS were the two active fractions responsible for its cardioprotective effect. The mechanism might involve protections of the cell membrane from hypoxic damage and of the cells from oxidative injury via preventing increased oxidative stress. Protection of the cells via inhibition of cellular apoptosis may also be involved. (C) 2013 Elsevier Ireland Ltd. All rights reserved.