摘要
目的 明确二肽基肽酶-4(DPP4)在泡球蚴感染所致肝纤维化中的作用。方法 取泡球蚴不同感染时期(1、3、6个月)小鼠肝脏组织,HE染色检查肝脏组织病理学变化,天狼星红染色法检测肝纤维化程度,免疫组织化学法检测α-平滑肌肌动蛋白(α-SMA)和DPP4的表达,采用相关分析法分析DPP4蛋白表达水平与肝纤维化的相关性。小鼠肝星状细胞(JS1),经60μg/ml泡球蚴蛋白(EmP)刺激后采用实时荧光定量PCR检测DPP4、α-SMA、COL1A1、TIMP1、MMP2的mRNA表达水平。结果 与对照组比较组,模型组小鼠感染泡球蚴后1、3、6个月α-SMA表达上调(阳性面积分别为3.521±0.8862、7.846±0.9873、15.34±0.6263)(均P<0.05),且呈时间依赖;天狼星红染色病灶旁纤维组织阳性面积增加(阳性面积分别为56979±9550、69844±763.8、82687±13774)(均P<0.05),且呈时间依赖;DPP4表达上调(阳性面积分别为5038±201.2、6110±174.4、9021±697.4)(均P<0.05),且呈时间依赖。泡球蚴感染小鼠DPP4表达水平与α-SMA相对表达量呈正相关(R^(2)=0.9166,P<0.05),与天狼星红阳性区域面积呈正相关(R^(2)=0.5368,P<0.05)。泡球蚴蛋白刺激小鼠肝星状细胞系JS1,肝星状细胞活化标志物α-SMA、COL1a1、TIMP1、MMP2 mRNA表达上调,DPP4的mRNA也随之上调(均P<0.05)。结论 DPP4参与泡球蚴感染所致肝纤维化过程并随纤维化加重表达上调,为阐明泡型棘球蚴病的致病机制和寻找新的治疗靶点奠定了基础。
Objective To determine the role of Dipeptidyl-Peptidase Ⅳ(DPP4) in hepatic fibrosis induced by E. multilocularis infection. Methods C57BL/6 female mice aged from 8 to 10 weeks were selected, and they were adaptively raised in independent ventilatory cages(IVC) barrier for 1 week. They were randomly divided into sham operation group and model group. The suspension of E. multilocularis was injected through portal vein, 2000 per mouse, and the sham operation group was injected with normal saline.The liver tissues of mice infected with E. multilocularis at different stages(1,3 and 6 months) were collected, the histopathological changes of liver were examined by HE staining, the degree of liver fibrosis was detected by Sirius red staining, the expression of alpha smooth muscle actin(α-SMA) and DPP4 was detected by immunohistochemistry, and the correlation between the expression level of DPP4 and liver fibrosis was analyzed by correlation analysis. Mouse hepatic stellate cells(JS1) were stimulated by 60 μg/ml E. multilocularis protein(EmP) and 20 ng/ml TGF-β1,then cultured for 24 h, and RNA was extracted. The mRNA expression levels of DPP4,α-SMA,COL1A1,TIMP1 and MMP2 were detected by RT-qPCR. Results Compared with the control group, the expression of α-SMA in the model group was up-regulated at 1,3 and 6 months after being infected with E. multilocularis(the positive areas were 3.521±0.8862, 7.846±0.9873, 15.34±0.6263 respectively)(all P<0.05),and it was time-dependent;Sirius red staining increased the positive area of the fibrous tissue adjacent to the lesion(the positive areas were 56979±9550, 69844±763.8, 82687±13774,respectively)(all P<0.05),which was time-dependent;The expression of DPP4 was up-regulated(positive areas were 5038±201.2, 6110±174.4, 9021±697.4,respectively)(all P<0.05),and it was time dependent. The expression level of DPP4 in mice infected with E. multilocularis was positively correlated with the relative expression of α-SMA(R^(2)=0.9166,P<0.05),and positively correlated with Sirius red positive area(R^(2)=0.5368,P<0.05). Mouse Hepatic stellate cells(JS1) stimulated by EmP,the mRNA expression of hepatic stellate cell activation markers α-SMA,COL1a1,TIMP1,MMP-2 was up-regulated, and the mRNA of DPP4 was also up-regulated(all P<0.05). Conclusion DPP4 is involved in the process of liver fibrosis caused by E. multilocularis and its expression is up-regulated with the aggravation of liver fibrosis, which lays a foundation for clarifying the pathogenesis of alveolar echinococcosis and finding new therapeutic targets.
作者
努尔拜提·库苏曼
木克西娜·木拉提
毕晓娟
杨宁
楚瑨
刘辉
房彬彬
吕国栋
李亮
张雪
孙立
林仁勇
Nuerbaiti Kusuman;Mukexina Mulati;BI Xiao-juan;YANG Ning;CHU Jin;LIU Hui;FANG Bin-bin;LV Guo-dong;LI Liang;ZHANG Xue;SUN Li;LIN Ren-yong(Department of Biochemistry and Molecular Biology,College of Basic Medicine,Xinjiang Medical University,Urumqi 830011,China;State Key Laboratory of Pathogenesis.Prevention,and Treatment of Diseases Highly Endemic to Central Asia,Clinical Medical Research Institute,The First Hospital Af-filiated with Xinjiang Medical University)
出处
《中国病原生物学杂志》
CSCD
北大核心
2023年第1期47-51,共5页
Journal of Pathogen Biology
基金
国家自然科学基金项目(No.82060371,81860361,32060223)
新疆维吾尔自治区重点实验室开放课题项目(No.2020D04028)
省部共建中亚高发病成因与防治国家重点实验室开放课题项目(No.SKL-HIDCA-2019-22,SKL-HIDCA-2021-30)。
关键词
多房棘球蚴
二肽基肽酶-4
肝纤维化
Echinococcus multilocularis
Dipeptidyl-Peptidase 4
liver fibrosis
作者简介
努尔拜提·库苏曼(1996-),女,新疆哈密人,在读硕士,从事棘球蚴病分子致病机制研究。E-mail:1779715046@qq.com;通讯作者:林仁勇,E-mail:renyonglin@xjmu.edu.cn
