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目的探讨Toll样受体4(TLR4)是否参与实验脑型疟的发生。方法采用伯氏疟原虫ANKA株(P.bANKA),以三种方式感染TLR4-/-C57BL/6小鼠和对照WT C57BL/6小鼠,分别为:模拟自然感染的按蚊叮咬、子孢子定量感染以及红内期疟原虫感染。按蚊叮咬感染和子孢子定量感染:复苏液氮冻存P.b ANKA原虫一管,腹腔注射2只C57BL/6小鼠,待小鼠原虫率增殖至5%~10%时取全血传至6只C57BL/6小鼠,供3~5日龄斯氏按蚊血餐,按蚊血餐19 d后,捆绑TLR4-/-C57BL/6小鼠和WT C57BL/6小鼠供按蚊叮咬感染,或者抓取感染性按蚊解剖出蚊唾液腺获得子孢子,然后定量感染实验小鼠;红内期原虫感染:液氮复苏P.b ANKA原虫,腹腔接种2只C57BL/6小鼠,3~4 d后取106感染疟原虫红细胞腹腔接种感染实验小鼠。结果三种感染方式下,在原虫率上,TLR4-/-C57BL/6小鼠和WT C57BL/6小鼠原虫生长曲线接近,说明TLR4缺失后不影响P.b ANKA原虫在小鼠体内的增殖;在实验脑型疟发生率和存活率上,TLR4-/-C57BL/6小鼠和WT C57BL/6小鼠均在5~10 d内全部发生脑型疟并死亡,脑型疟发生率均为100%,存活率均为0%。两组实验小鼠在原虫增殖、脑型疟发生率及存活率上均无差别。结论 TLR4不参与实验脑型疟的发生发展。
Objective To investigate whether Toll-like receptor 4 (TLR4) is involved in the pathogenesis of experimental cerebral malaria. Methods The P. clarkii Plasmodium ANKA strain (P.bANKA) was used to infect TLR4 - / - C57BL / 6 mice and control WT C57BL / 6 mice in three ways: anopheles bite simulating natural infection, Quantitative infection and red-phase Plasmodium infection. Anopheles bite infection and sporozoites quantitative infection: recovery of liquid nitrogen frozen Pb ANKA protozoa tube, intraperitoneal injection of two C57BL / 6 mice until the parasite multiplication rate of 5% to 10% of the whole blood to 6 C57BL / 6 mice were challenged with An. Mosquito bites for 3 to 5 days after blood meal and Anopheles mosquitoes blood meal was administered for 19 days. Then TLR4 - / - C57BL / 6 mice and WT C57BL / Or seizure infected Anopheles mosquitoes salivary glands dissected to obtain sporozoites, and quantitative infection of experimental mice; red protozoan infection: liquid nitrogen resuscitation Pb ANKA protozoa, intraperitoneal inoculation of 2 C57BL / 6 mice, 3 ~ 4 days after The infected mice were infected intraperitoneally with 106 malarial parasite erythrocytes. Results The growth curves of protozoal in TLR4 - / - C57BL / 6 and WT C57BL / 6 mice were close at the protozoal rate under the three infection modes, indicating that the deletion of TLR4 did not affect the proliferation of Pb ANKA protozoa in mice. In the experiment of cerebral malaria incidence and survival rate, all of TLR4 - / - C57BL / 6 mice and WT C57BL / 6 mice all died from cerebral malaria within 5 ~ 10 days. The incidence of cerebral malaria was 100 %, Survival rate is 0%. Two groups of mice in the protozoan proliferation, incidence and survival of cerebral malaria no difference. Conclusion TLR4 is not involved in the development of experimental cerebral malaria.