CAJ | 학술논문

目的：探讨四氯化碳诱导肝纤维化早期大鼠肝窦毛细血管化的形成过程。方法：清洁级雄性SD大鼠采用随机数字表法随机分为2组：正常对照组（ N组，6只）和肝纤维化模型组（ M组，32只）。 M组大鼠腹腔注射50％四氯化碳蓖麻油混合液， N组大鼠腹腔注射生理盐水，剂量为2 mL/kg，每周2次，共4周。分别于造模第3天、1周、2周和4周处死大鼠，HE染色和Masson染色观察肝脏组织炎症及纤维化的改变，透射电镜观察肝窦内皮细胞（LSECs）窗孔与基底膜（BM）的改变，免疫组织化学检测LSECs 表面标志物CD31及基底膜成分IV型胶原（Col IV）和层黏连蛋白（LN）的改变。结果：HE及Masson染色显示四氯化碳造模4周早期肝纤维化已形成。肝组织透射电镜显示四氯化碳造模第3天后开始出现LSECs窗孔直径变小及数目减少，随着造模时间的延长，LSECs失窗孔现象逐步严重，至第4周时局部内皮下可见连续的基底膜。免疫组化染色显示LSECs表面标志物CD31表达随着LSECs窗孔数目的减少而逐渐增强；基底膜成分Col IV于造模第2周时表达开始显著增强并随着造模时间延长表达逐渐增强，LN于造模第4周时表达开始显著增强。结论：肝纤维化早期大鼠局部肝组织可见典型的肝窦毛细血管化形成；肝窦壁内LN沉积是肝窦毛细血管化时形成连续基底膜的关键因素。
목적：탐토사록화탄유도간섬유화조기대서간두모세혈관화적형성과정。방법：청길급웅성SD대서채용수궤수자표법수궤분위2조：정상대조조（ N조，6지）화간섬유화모형조（ M조，32지）。 M조대서복강주사50％사록화탄비마유혼합액， N조대서복강주사생리염수，제량위2 mL/kg，매주2차，공4주。분별우조모제3천、1주、2주화4주처사대서，HE염색화Masson염색관찰간장조직염증급섬유화적개변，투사전경관찰간두내피세포（LSECs）창공여기저막（BM）적개변，면역조직화학검측LSECs 표면표지물CD31급기저막성분IV형효원（Col IV）화층점련단백（LN）적개변。결과：HE급Masson염색현시사록화탄조모4주조기간섬유화이형성。간조직투사전경현시사록화탄조모제3천후개시출현LSECs창공직경변소급수목감소，수착조모시간적연장，LSECs실창공현상축보엄중，지제4주시국부내피하가견련속적기저막。면역조화염색현시LSECs표면표지물CD31표체수착LSECs창공수목적감소이축점증강；기저막성분Col IV우조모제2주시표체개시현저증강병수착조모시간연장표체축점증강，LN우조모제4주시표체개시현저증강。결론：간섬유화조기대서국부간조직가견전형적간두모세혈관화형성；간두벽내LN침적시간두모세혈관화시형성련속기저막적관건인소。
[ABSTRACT]AIM:Toexplorethedevelopmentofhepaticsinusoidalcapillarizationintheearlystageofliverfi-brosis induced by carbon tetrachloride (CCl4) in rats.METHODS:Clean SD rats were randomly divided into normal con-trol group (group N, n=6) and liver fibrotic model group (group M, n=32).The rats in group N were intraperitoneal in-jected with saline and the rats in group M were intraperitoneal injected with CCl 4(2 mL/kg, twice a week for 4 weeks).At the end of the 3rd day and the 1st, 2nd and 4th weeks, all rats were killed and then the samples were collected .The patho-logical changes in the livers were observed by HE staining and Masson straining .The development of hepatic sinusoidal capillarization was observed by transmission electron microscopy (TEM) and immunohistochemical staining .The cell sur-face expression of vascular endothelium-associated marker CD31, collagen type Ⅳ(Col IV) and laminin (LN) was deter-mined.RESULTS:HE and Masson staining showed the formation of liver fibrosis after treatment with CCl 4 for 4 weeks. TEM showed that the fenestrate diameter of liver sinusoidal endothelial cells (LSECs) grew down, the fenestrate numbers of LSECs were decreased along with the development of liver fibrosis , and the consecutive basement membrane was formed at the end of the experiment .The expression of CD31 was significantly increased along with the development of defenestration , and the expression of Col IV and LN was significantly increased after the treatment with CCl 4 for 2 weeks and 4 weeks , re-spectively .CONCLUSION:The typical hepatic sinusoidal capillarization was detected in the early stage of liver fibrosis , and the deposition of LN in the liver sinusoidal walls was the mainly factor of formation of the consecutive basement mem -brane .