感染期血清外泌体在伯氏疟原虫红内期肝损伤中的作用  

Effect of serum-derived exosomes from Plasmodium berghei ANKA-infected mice on liver injury in blood stage of malaria

作　　者：黄丽 张鑫 黎广智 王永飞[2] 李小波[1] 金小宝[1] 黄博 HUANG Li;ZHANG Xin;LI Guangzhi;WANG Yongfei;LI Xiaobo;JIN Xiaobao;HUANG Bo(Guangdong Provincial Key Laboratory of Pharmaceutical Bioactive Substances,Guangdong Pharmaceutical University,Guangzhou 510006,China;School of Life Science and Technology,Jinan University,Guangzhou 510632,China;School of Life Science and Biopharmaceuticals,Guangdong Pharmaceutical University,Guangzhou 510006,China)

机构地区：[1]广东药科大学广东省生物活性药物研究重点实验室,广东广州510006 [2]暨南大学生命科学技术学院,广东广州510632 [3]广东药科大学生命科学与生物制药学院,广东广州510006

出　　处：《广东药科大学学报》2022年第3期69-75,共7页Journal of Guangdong Pharmaceutical University

基　　金：国家自然科学基金(81702020);广东药科大学国家级大学生创新创业项目(201910573003)。

摘　　要：目的 探讨感染伯氏疟原虫(P. berghei)小鼠血清的外泌体对红内期肝组织病理损伤的影响。方法 采用血清型总外泌体分离试剂法富集感染伯氏疟原虫小鼠血清的外泌体,透射电镜和Western blot鉴定外泌体。选取雌性昆明小鼠48只,随机分为4组:正常对照组(8只)、外泌体组(8只,每天每只鼠经尾静脉注射50μg外泌体)、感染组[16只,每只鼠经腹腔注射10^(6)个感染疟原虫红细胞(iRBC)]及感染+外泌体组(16只,每只鼠感染10^(6)个iRBC,且每天每只鼠经尾静脉注射50μg外泌体)。检测各组小鼠的生存时间、红细胞原虫感染率和脑型疟发生率,H&E染色观察肝组织病理变化,免疫组化检测肝组织M1型巨噬细胞标记物(iNOS)表达,以及qPCR检测肝组织促炎症/抑炎症细胞因子的mRNA表达水平。结果 在感染后第7~9天,感染组和感染+外泌体组均产生脑型疟模型小鼠和肝组织出现明显病理损伤。与感染组相比,感染+外泌体组小鼠的半数生存期显著缩短(P<0.05),脑型疟发生率显著升高(P<0.05),肝组织病理损伤显著加重(P<0.05),M1型巨噬细胞标记物(iNOS)表达显著上调(P<0.05)。同时,尾静脉注射外泌体后,感染小鼠肝组织的促炎症细胞因子(TNF-α、IL-1β和IL-6)的mRNA水平显著上调(P<0.05),而抑炎症细胞因子(IL-4和IL-10)的mRNA水平显著下调(P<0.05)。结论 感染期血清外泌体可能通过促进肝组织巨噬细胞M1极化和诱导过度的促炎症反应,从而加重伯氏疟原虫红内期肝组织的病理损伤。Objective To investigate the effect of serum-derived exosomes from Plasmodium berghei ANKAinfected mice on liver injury in blood stage of malaria. Methods The exosomes were isolated from peripheral blood of Kunming mice infected with P. berghei ANKA by total exosome isolation kit, and characterized by TEM and western blot. A total of 48 mice were divided into 4 groups, including the control or exosome group(the uninfected mice received daily i. v. injection of PBS or exosomes), Pb or Pb+Exos group(the infected mice received daily i. v. injection of PBS or exosomes). The survival time, erythrocyte protozoa infection rate and cerebral malaria incidence of mice in each group were detected. The pathological changes of liver tissue were observed by H&E staining. The expression of M1 macrophage marker(iNOS) in liver tissue was detected by immunohistochemistry. The mRNA expression levels of pro-inflammatory/anti-inflammatory cytokines in liver tissue were detected by qPCR. Results On the 7th-9th day after infection, cerebral malaria model mice were detected in Pb and Pb+Exos groups, and showed severe damage in liver tissues. Compared with the infection group, the half survival time of mice in the infection+exosome group was significantly shortened, the incidence of cerebral malaria was significantly increased, the pathological injury of liver tissue was significantly aggravated,and the expression of M1 macrophage marker(iNOS) was significantly upregulated(P<0.05). Meantime, the levels of TNF-α, IL-1β and IL-6 mRNA in liver tissues were significantly increased, but the levels of IL-4 and IL-10 mRNA were decreased(P<0.05). Conclusion Serum exosomes during infection may aggravate the pathological damage of liver tissue in the blood stage of P. berghei ANKA by promoting M1 polarization of liver macrophages and inducing excessive pro-inflammatory response.

关 键 词：伯氏疟原虫 红内期 肝脏 外泌体 巨噬细胞M1极化 

分 类 号：R392.7[医药卫生—免疫学]