目的 以“醋制入肝”中药炮制理论为基础，采用HPLC法分析不同醋制时间五味子成分变化，探讨成分变化与抗肝损伤作用关联，明确醋五味子保肝活性物质基础。方法 应用HPLC法建立10批生五味子及醋制1、2、3、4、5、6 h五味子指纹图谱，检测五味子醇甲、五味子乙素、五味子甲素、丙烯酰胺、5-羟甲基糠醛（5-HMF）、五味子酯甲水平。以CCl 4诱导小鼠急性肝损伤为动物模型，结合血清生化指标天冬氨酸氨基转移酶（AST）、丙氨酸氨基转移酶（ALT）、总超氧化物歧化酶（T-SOD）、丙二醛（MDA）、肿瘤坏死因子-α（TNF-α）的变化，运用灰色关联度法联合偏最小二乘法分析其保肝作用成分与药效之间的关联性。结果 10批五味子指纹图谱中，以五味子醇甲色谱峰为参比峰，标定出共有峰23个，其中特征峰6个；对10批生五味子及不同醋制时间下五味子指纹图谱与对照指纹图谱相似度组内对比发现，相似度均在0.913～ 0.999；随着蒸制时间的延长，五味子中5-HMF整体呈上升趋势；醋制3 h时五味子酯甲较生五味子质量分数略有增加，而后随着时间延长逐渐减小；醋制1～6 h内五味子醇甲、五味子甲素和五味子乙素质量分数较生五味子变化不大。与模型组比较，醋制2、3、6 h组AST、ALT、TNF-α、MDA水平显著降低，SOD水平显著升高（P<0.01）；醋制1 h组AST、MDA水平显著降低，SOD水平显著升高（P<0.01）；醋制4 h组AST、ALT水平显著降低，SOD水平显著升高（P<0.01）；醋制5 h组AST、ALT、TNF-α水平显著降低，SOD水平显著升高（P<0.01）。组间对比发现，醋制3 h组药效学指标明显较好，随着炮制时间延长，与醋制3 h组比较，醋制4、5、6 h组血清中AST、ALT、TNF-α、MDA水平有不同程度升高，SOD含量下降。故以醋制3 h五味子指纹图谱与药效学之间关联性建立谱-效关系研究，经灰色关联度法联合偏最小二乘法分析，5-HMF、五味子乙素、五味子醇甲为改善小鼠肝损伤的主要贡献成分。结论 醋制3 h五味子对CCl4诱导急性肝损伤小鼠的作用效果最佳，其中五味子乙素、5-HMF及五味子醇甲对小鼠生化指标影响较显著。

Objective To investigate the composition changes of Schisandra chinensis with different vinegar preparation times, based on the traditional Chinese medicine processing theory of "vinegar preparation into liver", HPLC method was used to analyze the active ingredients in S. Chinensis with different vinegar preparation times, and identify the basis of the active substances protecting liver. Methods HPLC was used to establish 10 batches of raw S. chinensis and 1, 2, 3, 4, 5, and 6 h fingerprint spectra of S. chinensis prepared with vinegar. The levels of schisandra alcohol A, schisandra B, schisandra A, acrylamide, 5-hydroxymethylfurfural (5-HMF), and schisandra ester A were detected. Using CCl₄ induced acute liver injury in mice as an animal model. Combined with serum biochemical indicators such as aspartate aminotransferase (AST), alanine aminotransferase (ALT), total superoxide dismutase (T-SOD), malondialdehyde (MDA), and tumor necrosis factor-α (TNF-α), using the grey correlation method combined with partial least squares method to analyze the correlation between its liver protective components and drug efficacy. Results In the fingerprint spectra of 10 batches of S. chinensis, a total of 23 peaks were identified using the schisandra alcohol methyl chromatographic peak as the reference peak, including six characteristic peaks. Comparing the similarity between the fingerprint of S. chinensis from 10 batches of raw S. chinensis and the control fingerprint under different vinegar processing times, it was found that the similarity was between 0.913 and 0.999 within the group. With the extension of steaming time, the overall 5-HMF content in S. chinensis shows an upward trend. When vinegar was used for 3 h, the mass fraction of S. chinensis ester A slightly increased compared to raw S. chinensis, and then gradually decreased with time. Within 1—6 hours of vinegar production, the mass fractions of schisandrin A, schisandrin A, and schisandrin B showed little change compared to those of raw S. chinensis. Compared with model group, the MDA, AST, ALT, and TNF- α level in the vinegar treated groups for 2, 3, and 6 h significantly decreased, while the SOD level significantly increased (P< 0.01), the levels of AST and MDA in the 1 h vinegar group significantly decreased, while the levels of SOD significantly increased (P< 0.01), the levels of AST and ALT in the 4 h vinegar group were significantly reduced, while the levels of SOD were significantly increased (P< 0.01); the level of AST, ALT, TNF- α significantly decreased and the SOD level significantly increased in 5 h vinegar group (P< 0.01). Comparison between groups showed that the pharmacodynamic indicators of the vinegar treated 3-hour group were significantly better. As the processing time prolonged, compared with the vinegar treated 3-hour group, the AST, ALT, and TNF-α levels in the serum of the vinegar treated 4, 5, and 6 hour groups were increased to varying degrees, while SOD content decreased. Therefore, a spectrum effect relationship study was established based on the correlation between the 3-hour fingerprint of S. chinensis prepared with vinegar and its pharmacodynamics. According to the combination of grey correlation analysis and partial least squares analysis, 5-HMF, schisandrin B, and schisandrin A are the main contributing components to improve liver injury in mice. Conclusion The best effect of 3-hour vinegar preparation of S. chinensis on CCl₄ induced acute liver injury in mice was observed, with schisandra B, 5-HMF, and schisandra alcohol A having significant effects on biochemical indicators in mice.

R284.1

黑龙江省应用技术研究与开发计划重点项目（GA19C009）