基于中药血清药物化学及血清药理学方法探讨荭草保护心肌细胞氧化损伤的物质基础

目的:应用中药血清药物化学及血清药理学方法探讨荭草保护H9c2心肌细胞氧化损伤的物质基础。方法:通过比较荭草提取液、给药后含药血清和空白血清的超高效液相色谱(UPLC)指纹图谱,寻找荭草的入血成分;建立体外培养的H9c2心肌细胞H2O2氧化损伤模型,以荭草含药血清对氧化损伤心肌细胞存活率、乳酸脱氢酶(LDH)漏出率、丙二醛(MDA)含量、超氧化物歧化酶(SOD)活力的影响为评价指标,并采用吖啶橙(AO)和溴化乙锭(EB)双荧光染色法观察荭草含药血清对H2O2氧化损伤的心肌细胞形态学的影响,研究荭草含药血清对氧化损伤心肌细胞的保护作用。结果:灌胃给予荭草提取液后,从大鼠血清中发现25个移行成分,其中有9个为荭草中的原型成分,16个可能为其在体内转化的代谢产物;与模型组相比,荭草含药血清各剂量组能明显降低氧化损伤心肌细胞LDH漏出率、MDA含量(P<0.05),显著升高T-SOD活性和细胞存活率(P<0.05或P<0.01),有效抑制细胞凋亡。结论:荭草含药血清中的移行成分及其代谢产物可能是荭草保护心肌细胞氧化损伤的物质基础。

Research on Material Basis of Polygonum orientale Protecting H9c2 Cells against Oxidative Injury Based on Traditional Chinese Medicine Serum Chemical and Serum Pharmacological Methods

Objective: To investigate the material basis of Polygonum orientale protecting cardiac H9c2 cells against oxidative injury induced by hydrogen peroxide(H₂O₂) basing on traditional Chinese medicine serum chemical and serum pharmacological methods. Method: The absorbed components of P. orientale into blood were determined by comparing the ultra performance liquid chromatography(UPLC) fingerprints of the extract, medicinal serum and blank serum. Simultaneously, the oxidative injury models induced by H₂O₂ was established by cultivating cardiac H9c2 cells in vitro and then the cell viability, lactic dehydrogense(LDH) release, malondialdehyde(MDA) production as well as superoxide dismutase(SOD) activity were analyzed. Moreover, the morphology of cardiac H9c2 cells damaged by H₂O₂ were observed under an inverse fluorescence microscope after dyed by AO/EB. All of them were used to evaluate the protective effect of P. orientale on H₂O₂-induced injury. Result: Twenty five transitional components of P. orientale absorbed into blood after intragastrical administration were detected, nine of which were original components and sixteen are possible metabolites. Furthermore, compared with the model group, medicated serum decreased the release of LDH, MDA (P<0.05). However, it increased the activity of SOD and cell viability (P<0.05 or P<0.01), while apoptosis of cardiac H9c2 cells were obviously inhibited. Conclusion: The absorbed components into blood may be material basis of P. orientale used for protecting cardiac H9c2 cells against oxidative injury.