青蒿琥酯和双氢青蒿素对华支睾吸虫所致小鼠肝纤维化的治疗作用研究  

Therapeutic effects of artesunate and dihydroartemisinin on hepatic fibrosis induced by Clonorchis sinensis in mice

作　　者：汤宪时 梁幼生[1] 许永良[1] 吴潇潇 季文翔 仝德胜[1] TANG Xian-shi;LIANG You-sheng;XU Yong-liang;WU Xiao-xiao;JI Wen-xiang;TONG De-sheng(Key Laboratory of National Health and Family Planning Commission on Parasitic Disease Control and Prevention,Jiangsu Provincial Key Laboratory on Parasite and Vector Control Technology,Jiangsu Institute of Parasitic Diseases,Wuxi 214064,China)

机构地区：[1]国家卫生健康委员会寄生虫病预防与控制技术重点实验室,江苏省寄生虫与媒介控制技术重点实验室,江苏省血吸虫病防治研究所,无锡214064

出　　处：《中国人兽共患病学报》2022年第12期1098-1105,共8页Chinese Journal of Zoonoses

基　　金：江南大学公共卫生研究中心项目(No.JUPH201838)。

摘　　要：目的观察和评估青蒿琥酯(ART)和双氢青蒿素(DHA)对华支睾吸虫所致小鼠肝纤维化的治疗作用。方法BALB/c小鼠35只,每只小鼠灌胃感染30个华支睾吸虫囊蚴,饲养30 d,建立小鼠华支睾吸虫病肝纤维化模型,随机分为7组,即青蒿琥酯组(ART组)、氯化钠组(NaCl组)、吡喹酮组(PZQ组)、吡喹酮+青蒿琥酯组[(PZQ+ART)组]、双氢青蒿素组(DHA组)、聚氧乙烯蓖麻油组(EL组)、吡喹酮+双氢青蒿素组[(PZQ+DHA)组],每组5只小鼠。采用腹腔注射法(ART)和经口灌胃法(DHA),用药剂量都为20 mg/kg,每天1次,连续用药30 d,合并PZQ用药组于感染华支睾吸虫囊蚴30 d后300 mg/kg PZQ连续灌胃治疗2次。取各组小鼠肝组织,应用苏木精-伊红(HE)和马松(Masson)染色显示小鼠肝组织病理学变化和胶原增生情况,应用荧光定量PCR和Western Blot检测肝组织α-SMA、TGF-β、Col I、Col III mRNA和蛋白表达,应用肝功检测试剂盒测定小鼠血清中丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)和碱性磷酸酶(ALP)含量。结果HE和Masson染色显示小鼠感染华支睾吸虫囊蚴30 d后肝脏汇管区有炎性细胞浸润,周围出现大量胶原纤维,有肝纤维化形成;Western Blot结果显示,感染组小鼠肝脏Col III表达量(1.26±0.01)高于未感染组(0.75±0.14)(t=2.01,P<0.05)。荧光定量PCR结果显示,接受ART和DHA治疗的小鼠与各自对照组相比,各纤维化相关基因mRNA表达水平没有显著差异。Western Blot结果显示,接受ART治疗后,ART组(0.32±0.04、0.81±0.09)和(PZQ+ART)组(0.16±0.01、1.02±0.18)的Col I、Col III蛋白表达水平都低于NaCl组(1.54±0.38、2.24±0.35),而且PZQ+ART组的Col I表达水平低于ART组(t=1.98,P<0.05);接受DHA治疗的小鼠α-SMA、Col I蛋白表达水平与对照组比较没有显著差异。肝功能检测结果显示,ART组(197.75±8.84)U/L和(PZQ+ART)组(234.66±42.54)U/L血清AST水平与NaCl组(150±20.29)U/L比较明显升高;(PZQ+DHA)组(29±12.98)U/L血清ALT水平较EThis study was aimed at observing and evaluating the therapeutic effects of artesunate(ART)and dihydroartemisinin(DHA)on hepatic fibrosis induced by Clonorchis sinensis in mice.A total of 35 BALB/c mice were infected with 30 metacercariae each via intragastric administration,and a mouse model of clonorchiasis and liver fibrosis was established after 30 days.The mice were randomly divided into an ART group,sodium chloride(NaCl)group,praziquantel(PZQ)group,(PZQ+ART)group,DHA group,cremophor group(EL group)and(PZQ+DHA)group,with five mice each.ART(via intraperitoneal injection)and DHA(via intragastric administration)were both administered at a dosage of 20 mg/kg once per day,for 30 consecutive days.When ART was combined with PZQ,300 mg/kg PZQ was administered intragastrically twice on the 30th day after infection.HE and Masson staining were used to reveal histopathological changes and collagen hyperplasia in liver tissues of the mice in each group.The mRNA and protein expression ofα-SMA,TGF-β,Col I and Col III in the liver tissues was detected by fluorescence quantitative PCR and western blotting,respectively.The content of alanine aminotransferase(ALT),aspartate aminotransferase(AST)and alkaline phosphatase(ALP)in mouse serum were determined with liver function test kits.HE and Masson staining indicated that 30 days after infection,inflammatory cell infiltration appeared in the hepatic portal area,which was encircled by large amounts of collagen fibers.Western blot results indicated that the Col III expression in the liver in infected mice(1.26±0.01)was significantly higher than in uninfected mice(0.75±0.14)(t=2.01,P<0.05),thus further confirming the formation of liver fibrosis.Fluorescence quantitative PCR results indicated no significant difference in mRNA expression levels for each fibrosis related gene between mice treated with ART and DHA and mice in the corresponding control groups.Western blot results showed that after ART treatment,the Col I and Col III protein expression in the ART group(0.32±0.04,0.8

关 键 词：华支睾吸虫 肝纤维化 青蒿琥酯 双氢青蒿素 

分 类 号：R383.2[医药卫生—医学寄生虫学]