咔唑氨基醇化合物H1402对棘球蚴感染小鼠组织淋巴细胞及其亚群的影响

The impact of carbazole aminoalcohol H1402 on lymphocytes in mice infected with Echinococcus granulosus and Echinococcus multilocularis

目的 分析咔唑氨基醇化合物H1402对棘球蚴感染小鼠的淋巴细胞及其亚群的影响。方法 分别采集细粒棘球绦虫和多房棘球绦虫原头蚴,经肝门静脉接种建立感染小鼠模型。建模60 d后,分别用低剂量(50 mg/kg)和高剂量(80mg/kg)H1402每天灌胃干预两种棘球蚴病小鼠30 d。灌胃结束4周后留存肝组织观察病理变化,分离肝脏、脾脏和血液淋巴细胞,用流式细胞术检测小鼠组织淋巴细胞。结果 H1402治疗后棘球蚴感染小鼠肝脏病灶大小和数量减少,病理结果显示炎症反应减弱(P<0.05)。多房棘球蚴感染治疗组中,H1402高剂量组肝B细胞比例(24.35±7.46)%明显高于感染模型组(17.74±4.57)%(P<0.05);肝脏T细胞(1.15±1.14)%,包括Treg,CD4^(+)T,CD8^(+)T和活化的T细胞比例H1402高剂量组明显低于低剂量组(36.00±5.68)%(P<0.05)。脾脏T细胞比例H1402高剂量组(22.86±12.49)%显著低于低剂量组(46.29±7.22)%和感染模型组(42.16±7.44)%(P<0.05)。在血液中活化的T细胞比例,尤其是CD8^(+)T细胞比例H1402高剂量组(16.62±5.72)%显著低于低剂量组(34.63±5.06)%(P<0.05);B细胞比例尤其是活化的B细胞H1402低剂量组(0.32±0.06)%显著低于感染模型组(0.74±0.21)%(P<0.05)。细粒棘球蚴感染小鼠,肝脏T细胞,包括Treg和CD8^(+)T细胞均明显高于空白组,CD8^(+)T细胞比例H1402高剂量组(40.54±7.22)%显著高于模型组(9.88±5.93)%(P<0.05)。脾脏T细胞比例尤其是活化的T细胞均明显低于空白组(P<0.05)。血液淋巴细胞CD4^(+)T细胞比例H1402高剂量组(31.59±27.94)%显著降低(P<0.05)。结论 H1402对棘球蚴感染治疗有效,明显降低多房棘球蚴和细粒棘球蚴感染肝脏和脾脏的T细胞比例,多房棘球蚴感染小鼠药物干预后活化的CD4^(+)T细胞和活化的CD8^(+)T细胞比例上升,可能参与免疫调节,抑制多房棘球蚴的病理反应。

Objective To determine the efficiency of carbazole aminoalcohol H1402 against both cystic echinococcosis and alveolar echinococcosis and the impact of the component on lymphocytes in a mouse model infected with Echinococcus multilocularis and E.granulosus.Methods C57 mice were infected with protoscoleces of E.multilocularis and E.granulosus via hepatic portal vein.After 60 days of infection,50 mg/kg(low-dose,H1402-L) and 80 mg/kg(high-dose,H1402-H) of H1402 were orally given daily respectively for 30 days.The mice were rest for 4 weeks without any intervention before biopsy and the partial liver tissues were fixed for H&E staining.The livers,spleens and blood of the mice were collected for lymphocytes analysis by flow cytometry.Results H1402 treatment reduced the size and number of liver lesions in a dose-dependent pattern.H&E staining showed that H1402 decreased the inflammatory response in both E.multilocularis and E.granulosus lesions(P<0.05).H1402-H significantly induced a higher percentage of B cells(24.35±7.46)% compared to the cells in the infection control mice(17.74 4.57)%(P<0.05).However,the drug decreased liver T cells(1.15±1.14)%,including Treg,CD4^(+)T,CD8^(+)T and activated T cells(36.00±5.68)%(P<0.05).The proportion of spleen T cells including activated T cells in H1402-H group(22.86 12.49)% was significantly lower than that in the low-dose group(46.29 7.22)% and the infection control group(42.16±7.44)%(P<0.05).In blood profile analysis,the proportion of activated T cells,especially CD8^(+) cells in the H1402-H group(16.62±5.72)% was significantly lower than that in the low-dose group((34.63±5.06)%(P<0.05).B cells ratio,especially the activated B cells H1402-Lmice(0.32 0.06)% was significantly lower than the infection model group(0.74 0.21)%(P<0.05).In these mice infected with E granulosus,liver T cells,including Treg and CD8^(+)T cells were significantly higher than those in the blank group,and the proportion of CD8^(+)T cells in the H1402-H group(40.54±7.22)% was significantly higher than that in the model group(9.88±5.93)%(P<0.05).The proportion of spleen T cells,especially activated T cells,was significantly lower than that of the blank group(P<0.05).The proportion of CD4^(+)T cells in blood lymphocytes was significantly reduced in the H1402-H group(31.59±27.94)%(P<0.05).Conclusion H1402 is an effective drug for treating both E.granulosus and E.multilocularis infection in mice.After 30 days of treatment and 4 weeks of rest,T cells were significantly decreased in the liver and spleen of mice infected with E.multilocularis and E.granulosus.However,H1402 increases activated T cells likely associated with killing the larval parasite or directly function on the cells,which need further identification.