李文川++高良奎++李浩航++許佐明++羅宗將++汪建初++陸濤 浦澗

【摘要】目的探討熱缺血再灌注損傷（WIRI）早期大鼠肝臟基因表達(dá)譜的差異。

方法建立SD大鼠肝臟熱缺血再灌注損傷模型，取再灌注30 min和再灌注1 h肝組織標(biāo)本，采用基因表達(dá)譜芯片技術(shù)檢測各組基因表達(dá)的變化情況，確定差異表達(dá)基因。

結(jié)果篩選出大鼠肝臟再灌注30 min、再灌注1 h兩組共同差異表達(dá)基因有77條，其中表達(dá)上調(diào)56條，表達(dá)下調(diào)21條。 再灌注30 min或1 h表達(dá)上調(diào)的基因數(shù)均多于表達(dá)下調(diào)基因數(shù)，再灌注1 h表達(dá)發(fā)生變化的基因總數(shù)多于再灌注30 min。

結(jié)論大鼠肝臟熱缺血再灌注損傷伴隨多基因的表達(dá)譜改變，再灌注損傷早期的差異表達(dá)基因?yàn)檠芯扛闻KWIRI的分子機(jī)制提供實(shí)驗(yàn)依據(jù)。

【關(guān)鍵詞】肝臟；基因表達(dá)；再灌注損傷

中圖分類號(hào)：R657.3文獻(xiàn)標(biāo)識(shí)碼：ADOI：10.3969/j.issn.10031383.2017.01.006

A experimental study on gene expression profiles in rats liver at early phase of warm ischemia and reperfusion injury

[HJ1][HJ]

LI Wenchuan1，GAO Liangkui1，LI Haohang1，XU Zuoming1，LUO Zongjiang1，WANG Jianchu2，LU Tao2，PU Jian2▲

（1.Graduate School，2.Department of Hepatobiliary Surgery of Affiliated Hospital，Youjiang Medical University for Nationalities，Baise 533000，China）

[HJ2][HJ]

【Abstract】ObjectiveTo explore difference of gene expression profiles in rats liver at early phase of warm ischemia and reperfusion injury（WIRI）.

MethodsLiver WIRI model of SD rats was established，and liver tissue samples with 30 min and 1 h of reperfusion were taken.Gene expression microarray was used to detect changes of gene expression in each group，so as to identify differential expressed genes（DEGs）.

ResultsThe analysis data of the microarray showed that there were 77 common DEGs including 56 upregulated genes and 21 downregulated genes at both 30 min and 1 h of reperfusion.The number of upregulated genes at 30 min or 1 h of reperfusion was larger than that of downregulated genes.And the number of DEGs at 1 h of reperfusion was larger than that 30 min of reperfusion.

ConclusionWIRI of rats livers are always complicated with profile changes of multi gene expression，and differential expressed genes at early phase of reperfusion injury can provide experimental basis to molecular mechanism of liver WIRI.

【Key words】liver；gene expression；reperfusion injury

肝臟熱缺血再灌注損傷（warm ischemia and reperfusion injury，WIRI）是肝移植和肝臟切除術(shù)中出現(xiàn)的主要臨床問題[1]。其主要特征在于肝細(xì)胞損傷、無菌性炎癥及術(shù)后肝功能障礙[2]。研究表明[3～4]熱缺血誘導(dǎo)的活性氧（reactive oxygen species，ROS）生成及許多炎癥介質(zhì)的釋放有助于并加重再灌注早期誘導(dǎo)組織損傷。因此，本實(shí)驗(yàn)采用基因表達(dá)芯片技術(shù)對WIRI早期大鼠肝臟基因表達(dá)譜的改變進(jìn)行大規(guī)模、高通量、并行分析，初步篩選再灌注損傷早期調(diào)節(jié)其表達(dá)顯著的功能基因，并討論與以前研究的相似性和差異性，為進(jìn)一步闡明熱缺血再灌注損傷早期的分子調(diào)控機(jī)制提供理論基礎(chǔ)。

1材料與方法

1.1實(shí)驗(yàn)動(dòng)物模型的建立

SPF級(jí)雄性SpragueDawley（SD）大鼠16只，體重320～380 g，由右江民族醫(yī)學(xué)院動(dòng)物中心（許可證號(hào)：SCXK桂20120003）提供。術(shù)前禁食但可適量飲水。稱重后采用10%水合氯醛300 mg/kg腹腔注射麻醉，取腹部縱切口，離斷鐮狀韌帶及左右三角韌帶，分離肝胃韌帶，游離門靜脈及肝固有動(dòng)脈。3 min后將門靜脈和肝固有動(dòng)脈阻斷30 min，造成原位肝臟熱缺血模型，切取標(biāo)本，即熱缺血30 min組。再灌注損傷組為阻斷30 min后恢復(fù)肝臟血流后30 min、1 h分別切取標(biāo)本。假手術(shù)組僅常規(guī)進(jìn)腹后，游離但不予結(jié)扎肝十二指腸韌帶內(nèi)血管，30 min后切取標(biāo)本。