目的 探究蒺藜茎、叶、果中呋甾皂苷26-O-β-葡萄糖苷酶（furostanolglycoside 26-O-β-glucosidase，F26G）活性-呋甾皂苷-螺甾皂苷变化的相关性，明确4种皂苷类成分分布规律，并比较茎、叶、果的肝肾毒性。方法 采用对硝基苯-β-D-葡萄糖（4-nitrophenyl-β-D-glucopyranoside，pNPG）法测定7～11月的蒺藜茎、叶、果中F26G活性，采用UHPLC-MS/MS法测定其中4种皂苷类成分26-O-β-D-glucopyranosyl-(25R)-5α-furostan-12-one-3β,22α,26-triol-3-O-β-D-galactopyranosyl-(1→2)-[β-D-xylopyranosyl-(1→3)]-β-D-glucopyranosyl-(1→4)-β-D-galactopyranoside（FOT）、蒺藜皂苷D（terrestrosin D）、tribuluside A和25R-tribulosin的含量，将F26G活性与4种皂苷类成分的含量变化进行相关性分析。通过小鼠长期毒性实验，观察肝肾组织病理损伤情况，应用主成分分析比较茎、叶、果的肝肾毒性。结果 茎、叶、果中F26G活性随采集时间均呈先升后降的趋势，且为叶＞果＞茎。含量测定结果显示，FOT含量呈茎＞叶＞果的规律，蒺藜皂苷D含量在叶中最高，茎、果中接近，tribuluside A和25R-tribulosin含量均表现为叶＞茎＞果。相关性结果表明茎、叶、果中螺甾皂苷含量变化与F26G活性及呋甾皂苷的含量变化相关性均较强，提示F26G参与了螺甾皂苷的生成；茎、果中4种皂苷类成分含量变化的相关性强于叶、果中成分含量变化的相关性。小鼠长期毒性实验发现，与空白组比较，蒺藜茎、叶组小鼠肝肾功能指标显著升高，肝组织受损，伴有不同程度的炎性细胞浸润，肾组织可见肾小球肿大，而蒺藜果组肝肾组织受损程度较小。聚类结果显示叶肝毒性较大，茎肾毒性较大。结论 蒺藜茎、叶中螺甾皂苷成分含量高，肝肾毒性大；蒺藜果中螺甾皂苷成分含量低，肝肾毒性小，中医临床多以蒺藜果入药与其肝肾毒性较小有关。

Objective To explore the correlation between furostanolglycoside 26-O-β-glucosidase (F26G) activity-furostanol saponins-spirotanol saponins in the stems, leaves and fruits of Tribulus terrestris, and clarify the distribution of four saponins, and compare the hepatic-renal toxicity of the stems, leaves and fruits. Methods F26G activity in the stems, leaves and fruits of T. terrestris from July to November was determined by 4-nitrophenyl-β-D-glucopyranoside (pNPG) method. The contents of 26-O-β-D-glucopyranosyl-(25R)-5α-furostan-12-one-3β,22α,26-triol-3-O-β-D-galactopyranosyl-(1→2)-[β-D-xylopyranosyl-(1→3)]-β-D-glucopyranosyl- (1→4)-β-D-galactopyranoside (FOT), terrestrosin D, tribuluside A and 25R-tribulosin were determined by UHPLC-MS/MS. The correlation between F26G activity and the contents of four saponins was analyzed. Long-term toxicity tests of mice were carried out to observe the pathological damage of liver and kidney tissues. Principal component analysis was used to compare the hepatic-renal toxicity of stems, leaves and fruits. Results The activity of F26G in stems, leaves and fruits all showed a tendency of increasing and then decreasing with the collection time, and the order was leaves > fruits > stems. The results of content determination showed that FOT content was stems > leaves > fruits. The content of terrestrosin D was the highest in leaves, which was close to that in stems and fruits. The contents of tribuluside A and 25R-tribulosin was leaves > fruits > stems. The correlation results showed that changes in spirotanol saponins contents in stems, leaves and fruits correlated strongly with changes in both the activity of F26G and the contents of furostanol saponins, suggesting that F26G was involved in the production of furostanol saponins. The correlation between changes in the contents of the four saponins in stems and fruits was stronger than that between changes in the contents of the components in leaves and fruits. Long-term toxicity tests in mice showed that compared with blank group, the liver and kidney function indexes of T. terrestris stems and leaves group were significantly elevated, liver tissue was damaged with different degrees of inflammatory cell infiltration, and glomerulomegaly was seen in renal tissue, while the liver and kidney tissues were less damaged in the T. terrestris fruits group. The clustering results indicated greater hepatotoxicity for leaves and greater nephrotoxicity for stems. Conclusion The content of spirotanol saponins in the stem and leaves of T. terrestris is high, and it has high liver and kidney toxicity; The content of saponins in the fruit of T. terrestris is low, and its liver and kidney toxicity is small. In traditional Chinese medicine clinical practice, T. terrestris fruit is often used as a medicine due to its low liver and kidney toxicity.

R285.5

国家自然科学基金资助项目（82274102）；国家中医药管理局中药炮制技术传承基地项目（国中医药科技中药[2022]59号）；山东省自然科学基金资助项目（ZR2024MH318）；山东省2023年中医药高层次人才培育项目（鲁卫函[2023]143号）