[山豆根致肝损伤大鼠外周血中微小RNA早期变化特征的研究]

[Study on early change features of microRNA in the peripheral blood of Sophorae tonkinensis radix et rhizoma induced liver injury rats].

作者信息

Sheng Yun-hua, Jin Ruo-min, Yao Guang-tao, Zhou Lu, Qiao Jing-yi

机构信息

Research Center for Drug Safety Evaluation, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China.

出版信息

Zhongguo Zhong Xi Yi Jie He Za Zhi. 2013 Mar;33(3):385-91.

PMID:23713256

Abstract

OBJECTIVE

To study early change features of microRNA (miRNA) in the peripheral blood of Sophorae Tonkinensis Radix et Rhizoma induced liver injury rats, and to look for the miRNA biomarkers in the peripheral blood of early liver injury.

METHODS

Sixty Wistar rats were randomly divided into the control group and the Sophorae Tonkinensis Radix et Rhizoma (abbreviated as STRR) group, 30 in each group. Rats in the STRR group was administered with STRR decoction at 12 g/kg (2 mL/100 g), while equal volume of the distilled water was given to those in the control group. Rats were anesthetized on day 3, 7, 14, and 28, and 28 days after withdrawal. The serum samples were withdrawn. The alanine aminotransferase (ALT), aspartate transaminase (AST), total bile (TBIL), alkaline phosphatase (ALP), total protein (TP), and albumin (ALB) were detected. The globulin (GLO) level was calculated. HE staining was performed on the liver tissue to observe the pathomorphological changes. The whole blood was collected on day 7, 14, and 28 to perform the microarray test. The differentially expressed miRNAs were screened and verified by RT-PCR.

RESULTS

The ALT activity obviously increased on day 7 - 28 in the STRR group (P <0.05). The histopathological results showed the degeneration and swelling of the liver cells on day 28. In the microarray test, there were 11, 22, and 13 up regulated expressed miRNAs on day 7, 14, and 28, respectively. There were 1, 13, 2 down regulated expressed miRNAs on day 7, 14, and 28, respectively. By target gene prediction and pathway analysis of differentially expressed miRNA on day 7, 14, and 28, they involved in regulating and controlling signal transduction, cellular interaction, cytoskeleton. Differentially expressed miRNA might possibly participate in the process of liver injury. The RT-PCR result of the expression of miR-291a-5p with the peak time efficiency on day 7 showed that the expressions of miR-291a-5p in the peripheral blood and the liver tissue were basically identical.

CONCLUSION

miR-291a-5p could early indicate the liver injury, which could be taken as one of an early marker in STRR induced liver injury.

摘要

目的

研究山豆根致肝损伤大鼠外周血微小RNA（miRNA）的早期变化特征，寻找早期肝损伤外周血中的miRNA生物标志物。

方法

将60只Wistar大鼠随机分为对照组和山豆根（简称STRR）组，每组30只。STRR组大鼠按12 g/kg（2 mL/100 g）灌胃给予STRR水煎液，对照组给予等体积蒸馏水。分别于第3、7、14、28天及停药后28天对大鼠进行麻醉，采集血清样本，检测丙氨酸氨基转移酶（ALT）、天冬氨酸氨基转移酶（AST）、总胆汁（TBIL）、碱性磷酸酶（ALP）、总蛋白（TP）和白蛋白（ALB），计算球蛋白（GLO）水平。对肝组织进行HE染色，观察病理形态学变化。分别于第7、14、28天采集全血进行基因芯片检测，筛选差异表达的miRNA并通过RT-PCR进行验证。

结果

STRR组第7~28天ALT活性明显升高（P<0.05）。组织病理学结果显示第28天肝细胞出现变性、肿胀。基因芯片检测结果显示，第7、14、28天分别有11、22、13个上调表达的miRNA，分别有1、13、2个下调表达的miRNA。对第7、14、28天差异表达miRNA进行靶基因预测和通路分析，其涉及调控信号转导、细胞间相互作用、细胞骨架。差异表达的miRNA可能参与肝损伤过程。第7天miR-291a-5p表达达峰时效的RT-PCR结果显示，外周血与肝组织中miR-291a-5p表达基本一致。