目的 探究女贞子Ligustri Lucidi Fructus及其酒制品对雷公藤多苷致生殖毒性的减毒作用差异，并基于核因子E2相关因子2（nuclear factor erythroid 2-related factor 2，Nrf2）介导的睾丸抗氧化防御探究其减毒机制。方法 ICR小鼠连续28 d ig雷公藤多苷（40 mg/kg），同时ig女贞子或其酒制品进行干预。采用试剂盒检测血清中睾酮和睾丸组织中氧化应激相关指标的水平；采用苏木素-伊红（hematoxylin-eosin，HE）染色观察睾丸组织病理变化；采用Western blotting和qRT-PCR检测睾丸组织Nrf2、血红素加氧酶-1（heme oxygenase-1，HO-1）、还原型烟酰胺腺嘌呤二核苷酸磷酸醌脱氢酶1（nicotinamide adenine dinucleotide phosphate quinone oxidoreductase 1，NQO1）蛋白及mRNA表达。结果 雷公藤多苷致小鼠睾丸生精小管结构松散，生精细胞层次减少，成熟精子数量显著减少，睾酮水平显著降低（P＜0.01），睾丸脂质过氧化产物丙二醛（malondialdehyde，MDA）水平显著升高（P＜0.01）；女贞子（1、2 g/kg）能够显著抑制上述指标异常（P＜0.05、0.01），且女贞子（2 g/kg）还能够显著提升抗氧化酶活性（P＜0.05、0.01）；女贞子酒制品（1、2 g/kg）均能够显著抑制上述指标异常（P＜0.01），且女贞子酒制品（2 g/kg）能够更全面地逆转氧化应激失衡（P＜0.01），相同剂量下女贞子酒制品的减毒效果明显优于其生品（P＜0.05、0.01）。机制分析发现，雷公藤多苷显著下调小鼠睾丸组织Nrf2及其下游分子HO-1、NQO1的表达（P＜0.01），而女贞子及其酒制品（1、2 g/kg）能够显著上调Nrf2及其下游分子HO-1、NQO1的表达（P＜0.05、0.01），其中女贞子酒制品的作用显著优于女贞子（P＜0.01）。结论 女贞子及其酒制品均能够减轻雷公藤多苷引起的睾丸生殖毒性，且酒制品的减毒效果显著优于生品，其机制可能与增强Nrf2介导的睾丸抗氧化防御相关。

Objective To explore the differential attenuation effects of Nüzhenzi (Ligustri Lucidi Fructus) and its wine-processed product on Tripterygium wilfordii polycoride (TWP)-induced reproductive toxicity, and to investigate its attenuation mechanism based on nuclear factor erythroid 2-related factor 2 (Nrf2) mediated testicular antioxidant defense. Methods ICR mice were ig TWP (40 mg/kg) for 28 d, and intervention was also given with Ligustri Lucidi Fructus or its wine-processed products. Reagent kits were used to detect the levels of testosterone in serum and oxidative stress-related indicators in testicular tissue. Hematoxylin-eosin (HE) staining was used to observe pathological changes in testicular tissue. Western blotting and qRT-PCR were used to detect the protein and mRNA expressions of Nrf2, heme oxygenase-1 (HO-1) and nicotinamide adenine dinucleotide phosphate quinone dehydrogenase 1 (NQO1) in testicular tissue. Results TWP caused a loose structure of seminiferous tubules in testes of mice, a decrease in level of spermatogenic cells, a significant reduction in number of mature sperm, a significant decrease in testosterone levels (P < 0.01), and a significant increase in the level of malondialdehyde (MDA), a lipid peroxidation product in testes (P < 0.01). Ligustri Lucidi Fructus (1, 2 g/kg) could significantly inhibit the abnormal indicators mentioned above (P < 0.05, 0.01), and Ligustri Lucidi Fructus (2 g/kg) could also significantly enhance the activity of antioxidant enzymes (P < 0.05, 0.01). Ligustri Lucidi Fructus wine-processed products (1, 2 g/kg) could significantly inhibit the abnormal indicators mentioned above (P < 0.01), and Ligustri Lucidi Fructus wine-processed products (2 g/kg) could more comprehensively reverse oxidative stress imbalance (P < 0.01). At the same dose, the detoxification effect of Ligustri Lucidi Fructus wine-processed products was significantly better than that of their raw products (P < 0.05, 0.01). Mechanism analysis revealed that TWP significantly down-regulated the expressions of Nrf2 and its downstream molecules HO-1 and NQO1 in testicular tissue of mice (P < 0.01), while Ligustri Lucidi Fructus and its wine-processed products (1, 2 g/kg) significantly up-regulated the expressions of Nrf2 and its downstream molecules HO-1 and NQO1 (P < 0.05, 0.01), with Ligustri Lucidi Fructus wine-processed products showing a significantly better effect than Ligustri Lucidi Fructus (P < 0.01). Conclusion Ligustri Lucidi Fructus and its wine-processed products could alleviate the testicular reproductive toxicity caused by TWP, and the detoxification effect of wine-processed products is significantly better than that of raw products. The mechanism may be related to enhancing Nrf2 mediated testicular antioxidant defense.

R285.5

中原科技创新领军人才项目(254200510005); 河南省高校科技创新团队支持计划项目(26IRTSTHN020); 河南省自然科学基金青年科学基金项目(252300423825); 河南中医药大学2024年度研究生科研创新类项目(2024KYCX035)