目的 以“醋制入肝”中藥炮制理論為基礎(chǔ)，采用HPLC法分析不同醋制時間五味子成分變化，探討成分變化與抗肝損傷作用關(guān)聯(lián)，明確醋五味子保肝活性物質(zhì)基礎(chǔ)。方法 應(yīng)用HPLC法建立10批生五味子及醋制1、2、3、4、5、6 h五味子指紋圖譜，檢測五味子醇甲、五味子乙素、五味子甲素、丙烯酰胺、5-羥甲基糠醛（5-HMF）、五味子酯甲水平。以CCl 4誘導(dǎo)小鼠急性肝損傷為動物模型，結(jié)合血清生化指標(biāo)天冬氨酸氨基轉(zhuǎn)移酶（AST）、丙氨酸氨基轉(zhuǎn)移酶（ALT）、總超氧化物歧化酶（T-SOD）、丙二醛（MDA）、腫瘤壞死因子-α（TNF-α）的變化，運用灰色關(guān)聯(lián)度法聯(lián)合偏最小二乘法分析其保肝作用成分與藥效之間的關(guān)聯(lián)性。結(jié)果 10批五味子指紋圖譜中，以五味子醇甲色譜峰為參比峰，標(biāo)定出共有峰23個，其中特征峰6個；對10批生五味子及不同醋制時間下五味子指紋圖譜與對照指紋圖譜相似度組內(nèi)對比發(fā)現(xiàn)，相似度均在0.913～ 0.999；隨著蒸制時間的延長，五味子中5-HMF整體呈上升趨勢；醋制3 h時五味子酯甲較生五味子質(zhì)量分數(shù)略有增加，而后隨著時間延長逐漸減?。淮字?～6 h內(nèi)五味子醇甲、五味子甲素和五味子乙素質(zhì)量分數(shù)較生五味子變化不大。與模型組比較，醋制2、3、6 h組AST、ALT、TNF-α、MDA水平顯著降低，SOD水平顯著升高（P<0.01）；醋制1 h組AST、MDA水平顯著降低，SOD水平顯著升高（P<0.01）；醋制4 h組AST、ALT水平顯著降低，SOD水平顯著升高（P<0.01）；醋制5 h組AST、ALT、TNF-α水平顯著降低，SOD水平顯著升高（P<0.01）。組間對比發(fā)現(xiàn)，醋制3 h組藥效學(xué)指標(biāo)明顯較好，隨著炮制時間延長，與醋制3 h組比較，醋制4、5、6 h組血清中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五味子指紋圖譜與藥效學(xué)之間關(guān)聯(lián)性建立譜-效關(guān)系研究，經(jīng)灰色關(guān)聯(lián)度法聯(lián)合偏最小二乘法分析，5-HMF、五味子乙素、五味子醇甲為改善小鼠肝損傷的主要貢獻成分。結(jié)論 醋制3 h五味子對CCl4誘導(dǎo)急性肝損傷小鼠的作用效果最佳，其中五味子乙素、5-HMF及五味子醇甲對小鼠生化指標(biāo)影響較顯著。

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

黑龍江省應(yīng)用技術(shù)研究與開發(fā)計劃重點項目（GA19C009）