双氢青蒿素和咯萘啶复方配伍比例筛选与实验验证

Screening and verification of compatibility ratio between dihydroartemisinin and pyronaridine

目的筛选双氢青蒿素和咯萘啶组合物的最佳配伍剂量,观察该剂量下两种药物联合应用对疟疾小鼠的影响。方法以伯氏疟原虫感染C57小鼠建立疟疾模型,以小鼠感染率、抑制率和复燃率等为观察指标,将双氢青蒿素与咯萘啶的25个联合给药组用二元二次方程法和协同作用指数评价法(CI值法)进行剂量筛选。剂量优化后与青蒿琥酯-咯萘啶进行药效比较得到双氢青蒿素-咯萘啶最佳配伍剂量,并进行效价实验。取小鼠脑、肝脏HE染色进行病理检查,检测血清肿瘤坏死因子(TNF)-α、干扰素(IFN)-γ的表达水平。结果综合二元二次回归方程法和CI值法筛选得到双氢青蒿素-咯萘啶5个较优剂量组(mg·kg^(-1)):4/3、2/3、1/3、0.5/3、4/1.5,优化得到0.5/3、1/3和2/3三个剂量组,与青蒿琥酯-咯萘啶进行药效比较,确定双氢青蒿素-咯萘啶最优剂量组为1/3(mg·kg^(-1))。病理检查显示,双氢青蒿素-咯萘啶给药剂量为1/3(mg·kg^(-1))时,疟疾小鼠脑血管中未见感染的红细胞,肝脏组织排列整齐,无疟色素沉积,且血清中TNF-α、IFN-γ的表达水平均低于模型组(P<0.01)。结论双氢青蒿素1 mg·kg^(-1)和咯萘啶3 mg·kg^(-1)为最佳配伍剂量,联合应用可以明显降低伯氏疟原虫感染小鼠的感染率和复燃率,改善脑部微血管阻塞,并能有效清除肝脏中的疟色素沉积。

AIM To screen the optimal dose-combination of the composition of dihydroartemisinin and pyronaridine,and to observe the effect of the combined application of the two drugs on malaria rat.METHODS The malaria model was established in C57 mice infected with Plasmodium berghei,and the infection rate,inhibition rate and recrudescence rate of mice were used as the observation indicators.The 25 co-administration groups of dihydroartemisinin and pyronaridine were screened by binary quadratic equation method and synergy index evaluation method(CI value method).After the dose-combination was optimized,the effect was compared with artesunate-pyronidine to obtain the best dose-combination of dihydroartemisinin-pyronadine for potency experiment;the brain and liver of malaria mice were stained with HE for pathological examination and the serum tumor necrosis factor(TNF)-αand interferon(IFN)-γwere detected.RESULTS Five optimal dose-combination groups of dihydroartemisinin-pyronadine(mg·kg^(-1))4/3,2/3,1/3,0.5/3,4/1.5 were got by binary quadratic regression equation method and CI value method,and further optimization was done to obtain three dose groups 0.5/3,1/3 and 2/3.Then compared with the efficacy of artesunate-pyronidine,it was determined that the optimal dose-combination of dihydroartemisinin-pyronidine was 1/3(mg·kg^(-1)).The results of pathological examination staining showed that no infected red blood cells were seen in the cerebral blood vessels and the liver tissues were neatly arranged with no malaria pigment when the dose-combination of dihydroartemisinin-pyronaridine was 1/3(mg·kg^(-1)),and the expression levels of TNF-αand IFN-γin the serum were lower than those in the model group(P<0.01).CONCLUSION Dihydroartemisinin 1 mg·kg^(-1)and pyronaridine 3 mg·kg^(-1)is the optimal dose-combination,and the combined application can significantly reduce the infection rate and recrudescence rate of Plasmodium berghei infected mice,improve cerebral microvascular obstruction,and remove the malarial pigment deposition in the liver.