CAJ | 학술논문

目的：探讨慢病毒介导的脯氨酰寡肽酶（POP）过表达对硫代乙酰胺（TAA）诱导的大鼠肝纤维化模型的影响。方法40只雄性SD大鼠被随机分为4组，采用5％TAA诱导大鼠肝纤维化模型，各组分别在造模第1 w末门静脉注射生理盐水（正常对照组和TAA模型组）、空病毒（TAA模型+空病毒组）或携POP慢病毒（TAA模型+POP慢病毒组），在造模第4 w末处死动物，留取肝脏组织；采用苏木素-伊红（HE）染色及Masson染色观察肝脏病理形态学变化，采用化学比色法测定肝组织内羟脯氨酸含量，分别采用实时定量PCR或Western blot法检测POP mRNA表达或蛋白水平。结果模型组POP蛋白相对表达量较正常对照组水平显著降低（P<0.05），而POP转基因组POP蛋白水平较其余三组均显著升高（P<0.01）；POP mRNA相对表达变化与POP蛋白相一致。TAA模型组和空病毒组的纤维化评分多为Ⅱ级，肝纤维化半定量积分分别为（15.2±1.69）和（15.3±4.62），显著高于正常对照组（1.75土0.63）和POP慢病毒过表达载体组（7.75±2.71）（P<0.05）；模型组和空病毒组肝组织羟脯氨酸含量分别为（504.47±111.15）μg/g肝质量和（498.32±90.87）μg/g肝质量，均显著高于正常对照组[（298.20±47.47）μg/g肝质量，P<0.05]，而POP转基因组大鼠肝组织中羟脯氨酸含量为（383.52±43.49）μg/g肝质量，显著低于模型组和空病毒组（P<0.05）。结论肝纤维化时肝组织内POP水平下调，慢病毒载体可成功介导POP在大鼠肝组织内过表达，抑制TAA诱导的大鼠肝纤维化，降低肝组织羟脯氨酸水平。
목적：탐토만병독개도적포안선과태매（POP）과표체대류대을선알（TAA）유도적대서간섬유화모형적영향。방법40지웅성SD대서피수궤분위4조，채용5％TAA유도대서간섬유화모형，각조분별재조모제1 w말문정맥주사생리염수（정상대조조화TAA모형조）、공병독（TAA모형+공병독조）혹휴POP만병독（TAA모형+POP만병독조），재조모제4 w말처사동물，류취간장조직；채용소목소-이홍（HE）염색급Masson염색관찰간장병리형태학변화，채용화학비색법측정간조직내간포안산함량，분별채용실시정량PCR혹Western blot법검측POP mRNA표체혹단백수평。결과모형조POP단백상대표체량교정상대조조수평현저강저（P<0.05），이POP전기인조POP단백수평교기여삼조균현저승고（P<0.01）；POP mRNA상대표체변화여POP단백상일치。TAA모형조화공병독조적섬유화평분다위Ⅱ급，간섬유화반정량적분분별위（15.2±1.69）화（15.3±4.62），현저고우정상대조조（1.75토0.63）화POP만병독과표체재체조（7.75±2.71）（P<0.05）；모형조화공병독조간조직간포안산함량분별위（504.47±111.15）μg/g간질량화（498.32±90.87）μg/g간질량，균현저고우정상대조조[（298.20±47.47）μg/g간질량，P<0.05]，이POP전기인조대서간조직중간포안산함량위（383.52±43.49）μg/g간질량，현저저우모형조화공병독조（P<0.05）。결론간섬유화시간조직내POP수평하조，만병독재체가성공개도POP재대서간조직내과표체，억제TAA유도적대서간섬유화，강저간조직간포안산수평。
Objective To investigate the effects of lentivirus-mediated prolyl oligopeptidase overexpression on thioacetamide (TAA)-induced liver fibrosis in rats. Methods 40 male Sprague-Dawley (SD) rats were ran-domly divided into four groups. Thioacetamide (TAA,5%) was injected intraperitoneally to induce liver fibrosis in rats. After 1-week TAA/saline treatment,rats were given normal saline (normal control group and TAA model group),empty virus(TAA plus empty vector group),or lentivirus containing POP gene(lentivirus-POP)(TAA plus lentivirus-POP group);Rats were sacrificed 4 weeks after initial TAA/saline treatment;Pathological changes of the liver were examed by HE staining and Masson staining;Hydroxyproline content in liver tissues were detected by chemical colorimetric;Real-time PCR and Western blot were used to determinate POP mRNA expression and POP protein content in liver,respectively. Results Relative level of POP protein in model group was(0.123±0.04),sig-nificantly decreased compared with normal control [(0.189 ±0.052),P<0.05];POP protein level in rats received lentivirus-POP was(0.304±0.044),significantly higher than that of other groups(P<0.01);the changes in POP mR-NA expression was similar with POP protein content. The fibrosis grade in TAA model group and empty virus group were mostly of S2,and the semi-quanti-tative analysis of the fibrotic tissue by Masson staining were (15.2±1.69) and (15.3±4.62),respec-tively,which were significantly higher than those of the normal controls and TAA plus lentivirus-POP group [(1.75 土 0.63) and(7.75 ±2.71),P<0.05];Hy-droxyproline content in model group and empty virus group were[(504.47±111.15)μg/g wet liver weight] and[(498.32±90.87)μg/g wet liver weight],respectively, significantly higher than that of the normal controls [(298.20±47.47)μg/g wet liver weight,P<0.05],while hydrox-yproline content in TAA plus lentivirus-POP group was [(383.52±43.49)μg/g TAA wet liver weight],significantly lower than those of model group and empty virus group (P<0.05). Conclusions POP expression decreases in fi-brotic liver induced by TAA;POP can be successfully overexpressed in rat liver by lentivirus containing POP genes,attenuates TAA-induced liver fibrosis and decreases hydroxyproline content in the rat liver.