Expression and its significance of SphK-1 in liver tissue of mice with liver injury
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摘要: 目的 利用5%葡聚糖硫酸钠(dextran sulfate sodium,DSS)诱导建立小鼠肝损伤模型,检测肝组织中鞘氨醇激酶1(Sphingosine kinase-1,SphK1)的表达情况,探讨其与肝损伤的关系。 方法 40只SPF级Balb/C雄性小鼠随机分成4组:对照组和DSS模型组(细分为6周、8周、12周3个小组),每组10只。模型组用5%的DSS溶液按0.6 ml/(d·只)连续灌胃建模,并分别于6周、8周、12周后处死,改良赖氏法检测血清中谷丙转氨酶(ALT)含量,HE染色观察肝脏病理损伤,用于荧光定量PCR检测肝脏SphK1基因表达水平。 结果 与对照组比较,各模型组小鼠血清中ALT活性增强,2组比较差异具有统计学意义(P<0.05)。各模型组小鼠肝脏均有明显病理损伤,肉眼观,6周组最为明显;光镜下,各模型组均出现了大片的肝脏坏死,大量淋巴细胞浸润。各模型组的小鼠肝脏组织中SphK1的表达量下降,12周下降最为明显,与对照比较差异具有统计学意义(P<0.05),SphK1的表达量与血清中ALT含量基本成反比关系。 结论 DSS可诱导小鼠肝损伤模型,肝损伤后小鼠肝内SphK1也随之表达减少,肝损伤越严重SphK1表达越少,SphK1表达与肝脏损伤具有相关性。Abstract: Objective To establish the liver injury model in mice induced by 5% dextran sodium sulfate(DSS),detect the expression of sphingosine kinase-1(SphK-1) in liver of mice,and evaluate the relationship between the level of SphK-1 and hepatic injury. Methods Forty SPF Balb/C male mice were randomly divided into 4 groups:blank control group,DSS model group(subdivided into 6,8,12 weeks three sub-group),10 mice in each group.The hepatic injury mice model was induced by intragastric administration of 5% DSS[0.6ml/(d·mouse)].The improved Reitman method was used to detect the serum level of ALT.The hepatic pathological changes were tested by hematoxylin eosin(HE) stain.The expression of SphK-1 in the liver of mouse was detected by quantitative PCR. Results In comparing with control group,the serum levels of ALT was raised in each sub-group(P<0.05).The hepatic pathological injury was obviously in each sub-group,the most visible change by naked eye was observed in 6 weeks sub-group,the necrosis and lymphocytes infiltration were observed under light microscope.The expression of SphK-1 in liver was reduced in all three sub-groups,the most predominance reduction was observed in 12 weeks,the differences had statistical significance when compared with the control group(P<0.05).There was a substantially inversely proportional between the expression of SphK-1 and the content ALT in serum. Conclusion Hepatic injury mice model can be induced by 5% DSS.The SphK-1 expression level of liver was decreased along with the level of liver damage in sub-group.We presume that SphK-1 may be closely related with liver damage.
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Key words:
- Liver injury /
- Sph K-1 /
- Dextran sulfate sodium
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