Guo Yuan, Li Kai, Jiang Xiao-Hui, Yang Ting, Zheng Zhong-Yuan, Chen Li-Na, Li Yu-Jie, Zheng Xiao-Jun
College of Pharmacy, Shanxi Medical University, Taiyuan 030001, China.
Research Center of Artemisinin, China Academy of Chinese Medical Sciences, Beijing 100700, China.
Zhongguo Zhong Yao Za Zhi. 2018 Aug;43(16):3397-3403. doi: 10.19540/j.cnki.cjcmm.20180604.002.
Malaria is still the most severe strain of the human malaria parasites, and malaria disease is life-threatening which can result in severe anemia and cerebral malaria, especially in children in tropical Africa. Previous studies have shown that artemisinin and its derivatives could selectively kill erythrocytic stage of malaria and have a greater impact on the ring period. In recent years, there have been new findings of its mechanism continually. However, the concentration of artemisinin and its derivatives used in these studies can reach 50 to 80 times the half-inhibitory concentration in vitro. In this study, the international standard strain 3D7 of Plasmodium falciparum was used to culture in vitro. After half-inhibitory concentration of dihydroartemisinin was treated, the morphological changes of P. falciparum intraerythrocytic stage were observed, and then the 3D7 life cycle and effects of different developmental stages after dosing was explored. The 3D7 strain of P. falciparum was continuously synchronised more than 3 times. And dihydroartemisinin (DHA) at half maximal inhibitory concentration (10 nmol·L⁻¹) was administered for 6 hours after the last synchronization, and 3 life cycles were continuously observed (132 h). The results showed that compared with the parasites untreated by DHA, there was a noticeable delay in the life cycle of at least 36 h, indicating that the growth of 3D7 was significantly inhibited by DHA (<0.001), and the rate of ring formation was significantly reduced (<0.05). The trophozoites were abnormal in shape, such as shrink in size, and the number of merozoites in schizonts was significantly decreased (<0.05). These results suggested that non-killing concentrations of DHA (meaning parasites can be inhibited but not killed) can significantly inhibit the growth of P. falciparum, which may not only affect the ring stage, but also have an impact on other stages of the P. falciparum.
恶性疟原虫仍然是人类疟原虫中最严重的毒株,疟疾是危及生命的疾病,可导致严重贫血和脑型疟疾,尤其是在热带非洲的儿童中。先前的研究表明,青蒿素及其衍生物能够选择性地杀死疟原虫的红细胞内期,并且对环状体期有更大的影响。近年来,其作用机制不断有新的发现。然而,这些研究中使用的青蒿素及其衍生物的浓度可达到体外半数抑制浓度的50至80倍。在本研究中,使用恶性疟原虫的国际标准株3D7进行体外培养。用双氢青蒿素的半数抑制浓度处理后,观察恶性疟原虫红细胞内期的形态变化,然后探索给药后3D7的生命周期及不同发育阶段的影响。恶性疟原虫3D7株连续同步化3次以上。在最后一次同步化后,以半数最大抑制浓度(10 nmol·L⁻¹)的双氢青蒿素(DHA)给药6小时,并连续观察3个生命周期(132小时)。结果显示,与未用DHA处理的疟原虫相比,生命周期至少有36小时的明显延迟,表明DHA显著抑制了3D7的生长(<0.001),环状体形成率显著降低(<0.05)。滋养体形态异常,如体积缩小,裂殖体中的裂殖子数量显著减少(<0.05)。这些结果表明,非致死浓度的DHA(即能抑制但不能杀死疟原虫)可显著抑制恶性疟原虫的生长,这可能不仅影响环状体期,而且对恶性疟原虫的其他阶段也有影响。