p28GANK在肝再生大鼠中的基因和蛋白表达

Gene and protein expressions of p28GANK in rat with liver regeneration.

作者信息

Qin Jian-Min, Fu Xiao-Yong, Li Shen-Jing, Liu Shu-Qin, Zeng Jin-Zhang, Qiu Xiu-Hua, Wu Meng-Chao, Wang Hong-Yang

机构信息

International Co-operation Laboratory on Signal Transduction, Eastern Hepatobiliary Surgery Institute, Second Military Medical University, Shanghai, 200438, China.

出版信息

World J Gastroenterol. 2003 Nov;9(11):2523-7. doi: 10.3748/wjg.v9.i11.2523.

DOI:10.3748/wjg.v9.i11.2523

PMID:14606089

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC4656533/

Abstract

AIM

To observe the gene and protein expression changes of p28GANK in regenerating liver tissues, and to reveal the biological function of p28GANK on the regulation of liver regeneration.

METHODS

One hundred and thirty two adult male Sprague-Dawley rats were selected, weighing 200-250 g, and divided randomly into sham operation (SO) group and partial hepatectomy (PH) group. Each group had eleven time points: 0, 2, 6, 12, 24, 30, 48, 72, 120, 168 and 240 h, six rats were in each time point. The rats were undergone 70% PH under methoxyflurane anesthesia by resection of the anterior and left lateral lobes of the liver. SO was conducted by laparotomy plus slight mobilization of the liver without resection. Liver specimens were collected at the indicated time points after PH or SO. The expression level of p28GANK mRNA was determined by Northern blot as well as at protein level via immunohistochemical staining. The expressions of p28GANK mRNA in these tissues were analyzed by imaging analysis system of FLA-2000 FUJIFILM and one way analysis of variance. The protein expressions of p28GANK in these tissues were analyzed with Fromowitz' method and Rank sum test.

RESULTS

The expression of p28GANK mRNA in the regenerating liver tissues possessed two transcripts, which were 1.5 kb and 1.0 kb. There was a significantly different expression patterns of p28GANK mRNA between SO and PH groups (P<0.01). The expression of p28GANK mRNA increased 2 h after PH, the peak time was 72 h (SO group: 163.83+/-1.4720; PH group: 510.5+/-17.0499, P<0.01). There was a significant difference in the 1.5 kb transcript, which decreased gradually after 72 hours. The protein expression of p28GANK was mainly in the cytoplasm of regenerating hepatocytes, and increased near the central region 24 h after PH, and became strongly positive at 48 h (+++, vs the other time points P<0.05), but decreased 72 h after PH.

CONCLUSION

The expression of p28GANK mRNA increases in the early stage of rat liver regeneration, the protein expression of p28GANK is mainly in the cytoplasm of regenerating liver cells. It suggests that the gene of p28GANK may be an important regulatory and controlled factor involved in hepatocyte proliferation during liver regeneration.

摘要

目的

观察再生肝组织中p28GANK基因及蛋白表达变化，揭示p28GANK对肝再生调控的生物学功能。

方法

选取132只成年雄性Sprague-Dawley大鼠，体重200 - 250 g，随机分为假手术（SO）组和部分肝切除术（PH）组。每组有11个时间点：0、2、6、12、24、30、48、72、120、168和240 h，每个时间点6只大鼠。大鼠在甲氧氟烷麻醉下通过切除肝的前叶和左外叶进行70%肝切除术。假手术通过剖腹术加肝脏轻微移动但不切除来进行。在肝切除或假手术后的指定时间点收集肝脏标本。通过Northern印迹法测定p28GANK mRNA的表达水平，并通过免疫组织化学染色测定蛋白水平。用富士胶片FLA - 2000成像分析系统和单因素方差分析对这些组织中p28GANK mRNA的表达进行分析。用Fromowitz方法和秩和检验分析这些组织中p28GANK的蛋白表达。

结果

再生肝组织中p28GANK mRNA的表达有两种转录本，分别为1.5 kb和1.0 kb。SO组和PH组之间p28GANK mRNA的表达模式有显著差异（P<0.01）。肝切除后2 h p28GANK mRNA表达增加，峰值时间为72 h（SO组：163.83±1.4720；PH组：510.5±17.0499，P<0.01）。1.5 kb转录本有显著差异，72小时后逐渐下降。p28GANK的蛋白表达主要在再生肝细胞的细胞质中，肝切除后24 h在中央区域附近增加，48 h时变为强阳性（+++，与其他时间点相比P<0.05），但肝切除后72 h下降。