CAJ | 학술논문

目的研究肝细胞生长因子(HGF)与肝再生增强因子(ALR)重组表达质粒对大鼠实验性肝纤维化的治疗作用.方法建立二甲基亚硝胺肝纤维化模型后的90只S-D大鼠分为空白组、pcDNA3.1治疗组、pcDNA3.1-HGF治疗组、pcDNA3.1-ALR治疗组、pcDNA3.1-HGF与pcDNA3.1-ALR共治疗组、pcDNA3.1-HGF-ALR治疗组,每组15只.各治疗组大鼠每24小时经尾静脉注射1次相应质粒0.1 μmol,连续3次,空白组不接受任何治疗,另取同批次S-D大鼠10只作为对照组.治疗结束后4 d处死大鼠,留取肝组织采用HE染色观察肝脏的病理形态,采用免疫组织化学染色检测增殖细胞核抗原(PCNA)和c-jun的表达.计量资料采用单因素方差分析,两两比较采用SNK检验,计数资料采用Fisher确切概率法分析.结果空白组和pcDNA3.1治疗组大鼠肝组织有明显的条索状纤维间隔形成,可见假小叶,两组肝纤维化程度差异无统计学意义(x2=0.317,P=1. 000);其他治疗组肝纤维化均有不同程度改善,以pcDNA3.1-HGF-ALR治疗组改善最明显.对照组肝组织PCNA和c-jun表达均较低,其吸光度值分别为8.6±1.9和3.2±1.2;空白组与pcDNA3.1治疗组肝组织PCNA和c-jun表达均增加,其吸光度值分别为24.1±3.0.24.5±4.3与23.8±3.1、24.9±4.2,与对照组比较,差异有统计学意义(均P＜0.01);其他治疗组的PCNA表达显著增加,c-jun表达显著降低,均以pcDNA3.1-HGF-ALR治疗组的变化最明显.结论重组表达质粒pcDNA3.1-HGF-ALR能较好地改善大鼠实验性肝纤维化,并可能通过促进肝细胞增殖和抑制原癌基因c-jun的表达来发挥其抗肝纤维化作用.
목적 연구간세포생장인자(HGF)여간재생증강인자(ALR)중조표체질립대대서실험성간섬유화적치료작용.방법 건립이갑기아초알간섬유화모형후적90지S-D대서분위공백조、pcDNA3.1치료조、pcDNA3.1-HGF치료조、pcDNA3.1-ALR치료조、pcDNA3.1-HGF여pcDNA3.1-ALR공치료조、pcDNA3.1-HGF-ALR치료조,매조15지.각치료조대서매24소시경미정맥주사1차상응질립0.1 μmol,련속3차,공백조불접수임하치료,령취동비차S-D대서10지작위대조조.치료결속후4 d처사대서,류취간조직채용HE염색관찰간장적병리형태,채용면역조직화학염색검측증식세포핵항원(PCNA)화c-jun적표체.계량자료채용단인소방차분석,량량비교채용SNK검험,계수자료채용Fisher학절개솔법분석.결과 공백조화pcDNA3.1치료조대서간조직유명현적조색상섬유간격형성,가견가소협,량조간섬유화정도차이무통계학의의(x2=0.317,P=1. 000);기타치료조간섬유화균유불동정도개선,이pcDNA3.1-HGF-ALR치료조개선최명현.대조조간조직PCNA화c-jun표체균교저,기흡광도치분별위8.6±1.9화3.2±1.2;공백조여pcDNA3.1치료조간조직PCNA화c-jun표체균증가,기흡광도치분별위24.1±3.0.24.5±4.3여23.8±3.1、24.9±4.2,여대조조비교,차이유통계학의의(균P＜0.01);기타치료조적PCNA표체현저증가,c-jun표체현저강저,균이pcDNA3.1-HGF-ALR치료조적변화최명현.결론 중조표체질립pcDNA3.1-HGF-ALR능교호지개선대서실험성간섬유화,병가능통과촉진간세포증식화억제원암기인c-jun적표체래발휘기항간섬유화작용.
Objective To evaluate the therapeutic effects of recombinant expression plasmid containing hepatocyte growth factor (HGF) and augmenter of liver regeneration (ALR) on rats with hepatic fibrosis. Methods Ninety Sprague-Dawley rats, which had been established into hepatic fibrosis models, were equally divided into 6 groups: blank group, pcDNA3.1 therapy group,pcDNA3.1-HGF therapy group, pcDNA3. 1-ALR therapy group, pcDNA3.1-HGF and pcDNA3. 1-ALR combined therapy group, and pcDNA3. 1-HGF-ALR therapy group. Zero point one μmol of blank or plasmid was injected into model rats in each group by tail vein once a day for 3 days. Model rats in blank group didn't receive any treatment. Additional 10 rats were chosen as control group, which were not given any interference during the experiment. All rats were sacrificed 4 days after end of treatment. Liver tissues were reserved for observing pathologic changes after HE staining and detecting proliferating cell nuclear antigen (PCNA) and c-jun by immunohistochemistry. Measurement data were compared by single-factor analysis of variance. Comparison between groups was done by SNK test. Enumeration data were analyzed by Fisher's exact test. Results In blank group and pcDNA3.1 therapy group, hyperplasia of fibrous connective tissue was very obvious, false lobules were formed. There was no significant difference between these two groups (x2 =0. 317,P= 1. 000).In the 4 remaining groups, hepatic fibrosis all achieved different degree of amelioration, and the therapeutic effect of pcDNA3.1-HGF-ALR was optimal. In control group, the expressions of PCNA and c-jun in liver tissues were low, with absorbance value of 8.6±1.9 and 3.2 ± 1.2, respectively. In blank group and pcDNA3. 1 therapy group, the expressions of PCNA and c-jun were obviously increased, with absorbance value of 24. 1±3.0, 24.5±4.3 and 23.8±3.1, 24.9±4.2, respectively,which were significant different from control group (all P＜0.01). In the 4 remaining groups, the expressions of PCNA were all obviously increased, and expressions of c-jun were all obviously decreased. The maximum change scope was observed in pcDNA3. 1-HGF-ALR therapy group.Conclusions The recombinant expression plasmid pcDNA3. 1-HGF-ALR can effectively ameliorate experimental hepatic fibrosis of rats. The anti-fibrosis effects are achieved probably by up-regulating PCNA expression and down-regulating c-jun expression.