目的 建立灵芝菌与何首乌双向发酵的最佳发酵体系，并探讨灵芝菌生物转化对何首乌特异质肝毒性的影响。方法 70%乙醇回流提取生何首乌粉末制备何首乌醇提物（PMEE，二苯乙烯苷质量分数为7.85%）；HPLC法检测二苯乙烯苷转化率，考察底物浓度、发酵温度、转速、瓶装量、接种量、发酵时间6个因素对灵芝菌与PMEE双向发酵体系的影响，并制备不同二苯乙烯苷转化率的发酵物。采用脂多糖 （LPS） 制备 SD 大鼠特异质肝毒性模型，考察 PMEE 高、低剂量 （以生首乌计2.16、1.08 g·kg^-1，分别为4、2倍临床等效剂量）及灵芝菌转化后PMEE发酵物（二苯乙烯苷的转化率分别为0、50%、75%、100%，以生首乌计2.16 g·kg^-1）的特异质肝毒性，ELISA法检测血清丙氨酸氨基转氨酶（ALT）、天冬氨酸氨基转氨酶（AST）、乳酸脱氢酶（LDH）的释放量，HE染色法观察肝脏病理学改变。结果 灵芝菌转化PMEE的最佳发酵条件为发酵温度 28 ℃、摇床转速 180 r·min^-1、菌种接种量 2.5%、培养基瓶装量 25%、底物质量浓度 19.11 mg·mL^-1，转化时间 88～104 h，二苯乙烯苷转化率与转化时间呈一定的线性关系，线性方程为Y＝20.657X－12.959，R²＝0.988。与对照组比较，模型组的ALT、AST、LDH无显著性差异，提示LPS特异质肝毒性模型造模成功；与模型组比较，PMEE灵芝菌转化后二苯乙烯苷转化率为 75%、100% 组 ALT、AST、LDH 的释放量无显著差异，其他给 药 组 ALT、AST、LDH 释 放 量 均 显著升高（P＜0.01），且ALT、AST、LDH的释放量随二苯乙烯苷含量的减少而减少。与对照组相比，模型组有少量炎症因子增加，但无显著性病理学改变，未见肝损伤现象；PMEE高剂量组出现肝细胞广泛性坏死，胞核消失，有许多大小不一的脂滴空泡，偶有灶性炎症；PMEE低剂量组可见多数汇管区有大量库普弗细胞浸润，多数肝细胞水肿，有少量肝细胞胞质内可见小泡性或细颗粒状脂质空泡；PMEE灵芝菌转化组随着二苯乙烯苷含量降低，肝脏损伤程度也减轻，二苯乙烯苷转化率为100%组基本上未见肝细胞损伤。结论 灵芝菌生物转化能降低何首乌特异质肝毒性，减毒作用与减少何首乌中二苯乙烯苷含量相关。

Objective To establish of a fermentation system for Polygonum multiflorum Thunb. by Ganoderma lucidum, and effects of biotransformation of G. lucidum on idiosyncratic hepatotoxicity of P. multiflorum was investigated. Methods The ethanol extract of P. multiflorum (PMEE, mass fraction of stilbene glycoside 7.85%) was prepare by 70% ethanol reflux extraction of raw P. multiflorum powder. HPLC method was used to detect the stilbene glycoside conversion rate, and the effects of six factors, including substrate concentration, fermentation temperature, rotational speed, bottled volume, inoculation amount, and fermentation time, on the bidirectional fermentation system of G. lucidum and PMEE, and to prepare fermentation products with different stilbene glycoside conversion rates. Using lipopolysaccharide (LPS) to prepare a specific hepatotoxicity model in SD rats, the specific hepatotoxicity of high and low doses of PMEE (calculated as 2.16, 1.08 g·kg^-1, respectively, 4 and 2 times the clinical equivalent dose) and PMEE fermented products after transformation by G. lucidum (the conversion rates of stilbene glycosides are 0%, 50%, 75%, and 100%, respectively, 2.16 g·kg^-1 based on raw P. multiflorum) was investigated. The release of serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), and lactate dehydrogenase (LDH) were detected by ELISA, and the pathological changes of the liver were observed by HE staining. Results The optimal fermentation conditions for the transformation of PMEE by G. lucidum were fermentation temperature of 28 ℃ , rotating speed of the shaking table of 180 r·min^-1, inoculum inoculation amount of 2.5%, bottled amount of culture medium of 25%, and substrate mass concentration of 19.11 mg·mL^-1. There was a linear relationship between the conversion rate of stilbene glycoside and the conversion time within 88 to 104 hours, and the linear equation was Y=20.657 X－12.959, with a square of correlation coefficient R²=0.988. Compared with control group, there was no significant difference in ALT, AST, and LDH in the model group, indicating that the LPS specific hepatotoxicity model was successfully established. Compared with the model group, there was no significant difference in the release of ALT, AST, and LDH of PMEE fermented products after transformation by G. lucidum groups (the conversion rates of stilbene glycosides are 0%, 50%, 75%, and 100%), while the release of ALT, AST, and LDH in other treatment groups were significantly increased (P＜0.01), and the release of ALT, AST, and LDH decreased with the decrease of stilbene glycoside content. Compared with the control group, there was a small increase in inflammatory factors in the model group, but no significant pathological changes were observed, and no liver injury was observed. In the high dose group of PMEE, extensive necrosis of hepatocytes, disappearance of nuclei, many lipid droplet vacuoles of varying sizes, and occasional focal inflammation were observed. In the low dose group of PMEE, there were a large number of Kupffer cells infiltrating most portal areas, most liver cells were edema, and a small amount of vesicular or granular lipid vacuoles were visible in the cytoplasm of liver cells. With the decrease of the content of stilbene glycoside in the PMEE G. lucidum transformation group, the degree of liver damage was also reduced. The group with a stilbene glycoside conversion rate of 100% basically had no liver cell damage. Conclusion Biotransformation of G. lucidum can reduce heterologous hepatotoxicity of P. multiflorum. The toxicity reduction effect is related to reducing the content of stilbene glycosides in P. multiflorum.

湖南省卫生健康委科研计划项目（D202313058583）；湖南省大学生创新创业训练计划项目（湘教通【2022】174号：2924）；湖南中医药大学校级大学生创新创业训练计划项目（X202010541027）；湖南中医药大学校级科研基金项目（2020XJJJ020）