人参皂干Rg1对NOX3致斯氏狸殖吸虫肝纤维化的影响  

Effects of Rg1 on liver fibrosis caused by Pagumogonimus skrjabini via the NOX3 pathway

作　　者：戚健君 王文林[1] 李树德[2] QI Jian-jun WANG Wen-lin LI Shu-de(Department of Pathogen Biology, School of Basic Medical Sciences, Kunming Medical University, Kunming 650504, Yunnan , China Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Kunming Medical University, Kunming 650504, Yunnan , China)

机构地区：[1]昆明医科大学基础医学院病原生物学教研室,云南昆明650504 [2]昆明医科大学基础医学院生物化学分子生物学教研室

出　　处：《中国病原生物学杂志》2017年第5期385-388,393,共5页Journal of Pathogen Biology

基　　金：国家自然科学基金项目(No.81360252;81360128);云南省自然科学基金项目(No.2012FB025);云南公共卫生与疾病预防控制中心合作创新基金项目(No.2014YNPHXT01;2016YNPHXT05)

摘　　要：目的探讨人参皂干Rg1对NOX3在斯氏狸殖吸虫引起肝纤维化中的影响,以期发现潜在的治疗肝纤维化药物。方法 50只SD大鼠随机分为5组,每组10只。除正常组外,其他组每只大鼠腹腔注射感染斯氏狸殖吸虫囊蚴15只。8周后分别用低、中、高剂量Rg1溶液罐胃治疗该虫引起的肝纤维化,1次/d,连续8周。处死大鼠,取各组肝脏组织,用Western blot检测NOX3蛋白,qPCR检测NOX3mRNA含量;制备肝组织石蜡切片,采用免疫组化法检测肝细胞中NOX3蛋白的表达,分析Rg1对斯氏狸殖吸虫引起肝纤维化的作用。结果Western blot检测模型组大鼠NOX3含量较正常组升高(t=18.7,P<0.055,F=5.019,R square=0.9832);Rg1罐胃治疗组NOX3含量降低(P<0.01),以高剂量组降低更显著。低组(t=67.84,P<0.01,F=26.48,R square=0.9832),中组(t=55.41,P<0.01,F=4.269,R square=0.9980),高组(t=70.55,P<0.01,F=4.017,R square=0.8269)。qPCR检测模型组NOX3mRNA含量相对正常组升高(t=7.447,P<0.01,R square=0.9739),治疗组含量降低。低组(t=3.648,P<0.01,F=11.07,R square=0.9269),中组(t=5.558,P<0.01,F=4.679,R square=0.9374),高组(t=6.924,P<0.01,F=12.96,R square=0.8888)。免疫组化检测模型组NOX3高表达,肝细胞结构紊乱,可见成纤维细胞浸润。Rg1低、中、高剂量组治疗后NOX3依次降低,肝细胞形态好转。结论 NOX3在斯氏狸殖吸虫引起的肝纤维化中高表达,且在斯氏狸殖吸虫引起的肝纤维化中起重要作用。Rg1能在蛋白和mRNA水平上降低NOX3的表达,可作为潜在治疗斯氏狸殖吸虫引起的纤维化药物。Objective To examine the effects of ginsenoside Rg1 （Rg1） on liver fibrosis caused by Pagurnogonimus skrjabini in rats via the NOX3 pathway. P. skrjabini is found only in China, and in humans it is one of the main species causing extrapulmonary paragonimiasis. Hepatic fibrosis is the pathological basis for chronic liver disease. Blocking or re- versing liver fibrosis can prevent the development of liver cirrhosis. Rgl is the main active ingredient in ginseng and Pa- nax, and it has anti-oxidative and anti-fibrotic activities. Nevertheless, the molecular mechanisms by which Rgl alleviates hepatic fibrosis caused by P. skrjabini is still unclear. Methods Rats were infected with P. skrjabini to create a model of liver fibrosis. Groups were treated with different concentrations for eight weeks to observe the anti-fibrotic action of Rgl. Fifty Sprague-Dawley rats were randomly divided into 5 groups （n = 10）. Groups other than the normal group re- ceived an intraperitoneal injection of 15 P. skrjabini worms. Mice were then treated with a low, medium, or high dose of Rgl for 8 weeks. The rats were sacrificed and liver tissue was sampled. Real-time quantitative PCR and Western blotting were used to detect levels of NOX3 mRNA and protein. Pathological and morphological changes in the liver were observed using immunohistochemistry. Results Western blotting indicated that the expression of NOX3 in model groups was higher than that in the normal group （P〈0.05 t=18.75, F=5. 019, R square=O. 9832）. The expression of NOX3 decreased in rats receiving a low dose of Rgl （P-0.01, t--67.84, F=26.48, R square = 0. 9832）, rats receiving a medium dose of Rgl （P〈0.01, t=55.41, F=4.269, Rsquare=0.9980）, and rats receivinga high dose of Rgl （P〈0.01, t= 70.55, F=4. 017, R square：0.8269）. The level of NOX3 mRNA in model groups was higher than that in the normal group （P〈0.01, t 7. 447, R square=0.9739）. The level of NOX3 mRNA was higher in rats receiving a low dose of Rg 1 （ P〈0.01, t = 3. 648, F=11

关 键 词：斯氏狸殖吸虫 人参皂干Rg1 肝纤维化 NOX3 

分 类 号：R383.2[医药卫生—医学寄生虫学]