黄芪多糖对鲤化学性肝细胞损伤的抑制作用
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江苏省自然科学基金项目(BK2012535)、国家自然科学基金项目(31202002、31200918)和中央级公益性科研院所基本科研业务费专项(2013JBFM12)


Inhibition effect of Astragalus polysacharin on chemical hepatocyte damage of common carp (Cyprinus carpio)
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    摘要:

    为探讨黄芪多糖对鲤肝细胞的保护作用,利用8 mmol/L的四氯化碳(CCl4)作用肝细胞4 h来构建鲤肝细胞体外损伤模型,用不同质量浓度(0.2、0.4和0.8 mg/mL)的黄芪多糖分别前处理、后处理和前后处理肝细胞,然后观察肝细胞形态,检测肝细胞活力,细胞上清中谷草转氨酶(GOT)、谷丙转氨酶(GPT)、乳酸脱氢酶(LDH)、超氧化物歧化酶(SOD)和丙二醛(MDA)含量以及肝细胞中细胞色素P450酶1A和3A(CYP1A和CYP3A) mRNA的相对表达量。结果显示:黄芪多糖显著地抑制由CCl4损伤引起的肝细胞培养上清中GPT、GOT、LDH活性和MDA含量的升高,提高SOD活性及肝细胞的活力(P<0.05或P<0.01);同时,0.4和0.8 mg/mL黄芪多糖显著提高CYP1A mRNA的相对表达量(P<0.05),0.4 mg/mL黄芪多糖显著提高CYP3A mRNA 的相对表达量(P<0.05),形态学观察结果也显示黄芪多糖有效地抑制鲤肝细胞死亡。以上结果表明,黄芪多糖可有效保护鲤化学性肝细胞损伤,并对CYP1A和CYP3A的表达有积极作用。

    Abstract:

    To evaluate the inhibition effect of Astragalus polysacharin (APS) on chemical hepatocyte injury of common carp,APS (0.2,0.4 and 0.8 mg/mL) was added to the primary hepatocytes pre-treatment,post-treatment and both pre- and post-treatment with the incubation of hepatocytes in the culture medium with 8 mmol/L CCl4.The morphology of liver cells was then observed by microscope,lactate dehydrogenase (LDH),glutamate pyruvate transaminase (GPT),glutamate oxalate transaminase (GOT),superoxide dismutase (SOD) and malondial-dehyde (MDA) in the supernatants were measured,hepatocyte viability was investigated and the mRNA expression of cytochrome P450 1A and 3A (CYP1A and CYP3A) in the hepatocytes were studied.The results showed that APS significantly improved cell viability,inhibited the elevation of GPT,GOT,LDH and MDA,and increased the SOD level in the supernatants (P<0.05 or P<0.01).The expression of CYP1A mRNA was improved in 0.4 and 0.8 mg/mL APS treatments (P<0.05) and the expression of CYP3A mRNA was upgraded in 0.4 mg/mL APS treatment (P<0.05).The observation of morphology also showed that APS effectively inhibited mortality of the hepatocytes.It can be concluded from the data obtained that APS exhibited protective effect against CCl4-induced hepatotoxicity in fish,and positively induced the expression of CYP1A and CYP3A.

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贾睿,杜金梁,曹丽萍,徐跑,殷国俊,柏爱旭.黄芪多糖对鲤化学性肝细胞损伤的抑制作用[J].华中农业大学学报,2014,33(2):103-110

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  • 收稿日期:2013-04-28
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