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李漓,何雨晴,吴智煌,杨胜辉,周小江.巴豆叶乙酸乙酯提取物抗血吸虫活性的初步研究[J].湖南中医药大学学报英文版,2023,43(2):257-263.[Click to copy
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| 巴豆叶乙酸乙酯提取物抗血吸虫活性的初步研究 |
| 李漓,何雨晴,吴智煌,杨胜辉,周小江 |
| (湖南中医药大学, 湖南 长沙 410208) |
| 摘要: |
| 目的 研究巴豆叶乙酸乙酯提取物（ethyl acetate extract from the leaves of Croton tiglium, EAELCT）体内外抗血吸虫的活性及对血吸虫感染小鼠血清Th1/Th2型细胞因子分泌的调节作用。方法 体外抗血吸虫试验采用人工脱尾制备的血吸虫童虫，滴加于6孔培养板，45条童虫/孔，分为不同浓度EAELCT组、空白对照组、DMSO组以及吡喹酮组，比较各组童虫的死亡率；体内抗血吸虫试验采用血吸虫尾蚴腹部贴片法感染小鼠，建立血吸虫感染小鼠模型，感染后小鼠随机分为模型组、吡喹酮组、EAELCT高剂量组（0.3 g/kg）、EAELCT中剂量组（0.15 g/kg）和EAELCT低剂量组（0.075 g/kg）组。体内杀血吸虫童虫和成虫试验分别在感染后第1天和感染后第29天用不同剂量EAELCT连续灌胃7 d，并分别采集感染前、感染后第14天、第29天及感染后第21天、第42天的血液，分别于第29天和42天处死小鼠，收集肝门静脉和肠系膜静脉内的虫体，计算杀虫率。采用ELISA检测小鼠血清中IFN-γ、IL-2、IL-5、IL-13细胞因子的含量。收集左肝进行HE染色，镜下观察肝纤维化病变的程度，并通过Imagine-pro图像软件测定肝组织中虫卵肉芽肿面积。结果 体外试验显示，EAELCT对血吸虫童虫有明显杀灭效果，培养48 h和72 h后各浓度EAELCT组中血吸虫童虫的死亡率均显著高于空白对照组和吡喹酮组（P<0.05）。体内试验显示，与模型组相比，EAELCT各浓度组均显示较好的杀虫效果，每鼠虫荷数显著减少（P<0.05）；EAELCT各剂量组血清中Th1型细胞因子IFN-γ、IL-2的表达水平较模型组明显增加(P<0.05)，Th2型细胞因子IL-5、IL-13表达水平明显下降(P<0.05)；EAELCT各剂量组与模型组比较，肉眼可见小鼠肝脏的颜色较红润，肝脏表面的虫卵结节数目减少，HE染色显示肝脏虫卵肉芽肿面积下降明显（P<0.05）。结论 EAELCT体内外具有较好的抗血吸虫活性，可通过上调Th1型细胞应答和下调Th2型细胞应答，从而有效抑制血吸虫病肝纤维化的病变和程度。 |
| 关键词: 巴豆叶 乙酸乙酯提取物 抗血吸虫活性 调节细胞因子 血吸虫病肝纤维化 |
| DOI：10.3969/j.issn.1674-070X.2023.02.012 |
| Received:August 31, 2022 |
| 基金项目:湖南省自然科学基金项目（2020JJ4067）；湖南省教育厅科学研究项目（19A368）；湖南中医药大学学科开放基金项目（2018JCYX09）。 |
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| A preliminary study of the anti-schistosomiasis activity of ethyl acetate extract from the leaves of Croton tiglium |
| LI Li,HE Yuqin,WU Zhihuang,YANG Shenghui,ZHOU Xiaojiang |
| (Hunan University of Chinese Medicine, Changsha, Hunan 410208, China) |
| Abstract: |
| Objective To study the in vitro-in vivo antischistosomal activity of ethyl acetate extract from the leaves of Croton tiglium (EAELCT) and its regulatory effects on serum Th1/Th2 cytokine secretion in schistosomiasis-infected mice. Methods For in vitro anti-schistosomiasis test, schistosomula prepared by artificial tail removal were added dropwise to 6-well culture plates, 45 schistosomula/well, and were then divided into EAELCT groups with different concentrations, blank control group, DMSO group and group. The mortality rates of schistosomula in each group were compared. For in vivo anti-schistosomiasis test, schistosomiasis infected mouse model was established by schistosoma cercariae abdominal patch method. The infected mice were randomly divided into model group, praziquantel group, high-dose EAELCT group (0.3 g/kg), medium-dose EAELCT group (0.15 g/kg) and low-dose EAELCT group (0.075 g/kg). As to in vivo schistosomulum killing and schistosome killing tests, mice were continuously gavaged with different doses of EAELCT for 7 d on the first day and the 29th day after infection, and blood was collected before infection, 14, 29, 21 and 42 d after infection, and mice were sacrificed on the 29th and 42th day respectively, and worms in the portal vein and mesenteric vein were collected to calculate the insecticidal rate. The content of IFN-γ, IL-2, IL-5 and IL-13 cytokines in mice sera was detected by ELISA. The left liver was collected for HE staining, the degree of liver fibrosis lesions was observed microscopically, and the area of oval granulomas in liver tissue was determined by Imagine-pro. Results In vitro experiments showed significant killing effects of EAELCT on schistosomiasis, and the mortality rates of schistosomulum in the EAELCT group after 48 and 72 h culture were significantly higher than those in the blank control group and praz1quante group (P<0.05). In vivo experiments showed:compared with the model group, better insecticidal effects had been identified in EAELCT groups with all concentration, and the insect charge per mouse was significantly reduced (P<0.05); compared with the model group, the expression levels of Th1 cytokines IFN-γ and IL-2 in all EAELCT groups were significantly higher (P<0.05), and the expression levels of Th2 cytokines IL-5 and IL-13 were significantly lower (P<0.05); compared with the model group, the color of the mouse liver was rosier, the number of egg nodules on the surface of the liver was less, and the area of granuloma of liver eggs was reduced significantly in HE staining (P<0.05). Conclusion EAELCT has good antischistosomal activity both in vitro and in vivo, and can effectively inhibit the lesion and degree of schistosomiasis liver fibrosis by upregulating Th1 cell response and downregulating Th2 cell response. |
| Key words: leaves of Croton tiglium ethyl acetate extract antischistosomal activity regulation of cytokines schistosomiasis liver fibrosis |
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