从成分配伍和睾丸毒性探究“金钱草杀雷公藤毒”的相杀减毒机制

Detoxification mechanism of‘Lysimachiae Herba suppressing Tripterygium wilfordii toxicity’based on component combination and testicular toxicity

目的:基于成分配伍和睾丸毒性评价“金钱草杀雷公藤毒”的相杀减毒作用及机制。方法:基于成分配伍理念,以雷公藤的主要毒性成分雷公藤甲素(TP,0.4 mg/kg)以及金钱草的主要活性成分绿原酸(CA,20 mg/kg)、芦丁(Rut,12 mg/kg)、槲皮素(Que,10 mg/kg)为研究对象,分析比较TP与CA、Rut、Que、CARut、CA-Que、Rut-Que、CA-Rut-Que配伍前后对雄性小鼠睾丸毒性敏感指标血清睾酮和睾丸组织病理的影响,并采用蛋白免疫印迹法等检测p38 MAPK/ERK1/2/Nrf2/HO-1信号通路的关键分子。结果:TP灌胃14 d后显著降低小鼠睾丸系数和血清睾酮水平,使睾丸生精细胞大量坏死和核碎裂溶解,显著上调p38 MAPK并下调ERK1/2/Nrf2/HO-1通路,降低抗氧化物如超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、还原型谷胱甘肽(GSH)、谷胱甘肽转移酶(GST)、谷胱甘肽过氧化物酶(GPX)的水平(P<0.01),升高过氧化物丙二醛(MDA)的水平(P<0.01)。与TP单用比较,配伍CA-Rut、CA-Que、Rut-Que、CA-Rut-Que后不同程度上削弱了TP睾丸毒性,并不同程度逆转了p38 MAPK/ERK1/2/Nrf2/HO-1信号通路蛋白分子、抗氧化和过氧化物水平的异常(P<0.01,P<0.05)。结论:金钱草主要活性成分CA、Rut、Que的互相组合对雷公藤主要毒性成分TP诱导的睾丸毒性具有不同程度的相杀减毒作用,其中三者组合后的减毒作用最强;“金钱草杀雷公藤毒”的相杀减毒机制可能与成分配伍后调控p38 MAPK/ERK1/2/Nrf2/HO-1信号通路从而增强睾丸的抗氧化防御、减轻细胞坏死和核碎裂溶解有关。

Objective:To evaluate the detoxication effect and mechanism of‘Lysimachiae Herba suppressing Tripterygium wilfordii toxicity’based on component combination and testicular toxicity.Methods:Based on the component combination,taking the main toxic component of Tripterygium wilfordii,triptolide(TP,0.4 mg/kg)and the main active components of Lysimachiae Herba,including chlorogenic acid(CA,20 mg/kg),rutin(Rut,12 mg/kg),and quercetin(Que,10 mg/kg)as the objects,the serum testosterone and testicular pathological tissue before and after TP in combination with CA,Rut,Que,CA-Rut,CA-Que,Rut-Que,and CA-Rut-Que were detected and oberved.The key molecules in the p38 MAPK/ERK1/2/Nrf2/HO-1 signaling pathway were analyzed and compared by Western blotting and multiple kit methods.Results:Gavage of TP for 14 days caused a significant decrease in the testicular coefficient and serum testosterone of mice,massive necrosis of testicular spermatogenic cells and fragmentation and dissolution of nuclei,up-regulation of p38 MAPK,down-regulation of ERK1/2/Nrf2/HO-1 pathway,and an decreased level of antioxidant molecules such as superoxide dismutase(SOD),catalase(CAT),reduced glutathione(GSH),glutathione transferase(GST),and glutathione peroxidase(GPX),and an increased level of lipid peroxidation product malondialdehyde(MDA)(P<0.01).Compared with TP alone,TP in combination with CA-Rut,CA-Que,Rut-Que,and CA-Rut-Que weakened the testicular toxicity of TP and reversed the abnormal levels of MAPK/ERK1/2/Nrf2/HO-1 signaling pathway protein molecules,antioxidants,and peroxides induced by TP to varying degrees(P<0.01,P<0.05).Conclusion:Combination with CA,Rut and Que shows different degrees of detoxication on the testicular toxicity induced by TP,and Rut-Que-CA has the strongest detoxication effect;the detoxication mechanism of‘Tripterygium wilfordii in combination with Lysimachiae Herba’may be related to TP in combination with CA,Rut,and Que regulating the p38 MAPK/ERK1/2/Nrf2/HO-1 signaling pathway,thereby enhancing the antioxidant defense of the testis,reducing cell necrosis and nuclear fragmentation and lysis.