肝脏
肝髒
간장
CHINESE HEPATOLOGY
2014年
11期
839-843
,共5页
郭杞兰%黄月红%陈治新%王小众
郭杞蘭%黃月紅%陳治新%王小衆
곽기란%황월홍%진치신%왕소음
大鼠%肝纤维化%肝窦毛细血管化%肝窦内皮细胞%基底膜
大鼠%肝纖維化%肝竇毛細血管化%肝竇內皮細胞%基底膜
대서%간섬유화%간두모세혈관화%간두내피세포%기저막
Rats%Hepatic fibrosis%Hepatic Sinusoidal capillarization%Liver sinusoidal endothelial cells%Basement membrane
目的：观察四氯化碳（CCl4）诱导的大鼠肝纤维化过程中肝窦毛细血管化的形成过程，探讨其与肝纤维化的关系。方法32只清洁级雄性SD大鼠，随机分为正常对照组，肝纤维化模型组，正常对照组大鼠腹腔注射0．9％氯化钠溶液2 mL/kg ，模型组大鼠腹腔注射50％ CCl4-蓖麻油混合液2 mL/kg ，每周2次，共8周；分别于造模第2、4、6、8周处死大鼠，观察肝组织炎症及纤维化程度，放射免疫法检测血清中透明质酸（HA）的含量，透射电镜观察肝窦窦壁结构，S-P 免疫组织化学检测各组大鼠肝组织CD31、层黏连蛋白（LN）、IV型胶原（Col-IV）的表达。结果肝脏组织学证实CCl4诱导的大鼠肝纤维化模型构建成功，6周可见纤维间隔形成；透射电镜显示，CCl4诱导2周时，部分肝窦内皮细胞（liver sinusoidal endothelial cells ，LSEC）窗孔减少，内皮下未见基底膜（Basement membrane，BM），随着造模的进程，LSEC 窗孔进一步减少，部分甚至消失，第6、8周时局部肝窦内皮下形成连续的BM。同时，随着肝纤维化的进程，HA浓度逐渐升高，肝窦内皮细胞表面标志物CD31及基底膜主要成分Col-IV、LN表达逐渐增强。结论在CCl4诱导大鼠肝纤维化过程中，肝窦毛细血管化是逐渐形成的，LSEC失窗孔早于纤维间隔的形成，而肝窦内皮下基底膜出现在纤维间隔形成以后。
目的：觀察四氯化碳（CCl4）誘導的大鼠肝纖維化過程中肝竇毛細血管化的形成過程，探討其與肝纖維化的關繫。方法32隻清潔級雄性SD大鼠，隨機分為正常對照組，肝纖維化模型組，正常對照組大鼠腹腔註射0．9％氯化鈉溶液2 mL/kg ，模型組大鼠腹腔註射50％ CCl4-蓖痳油混閤液2 mL/kg ，每週2次，共8週；分彆于造模第2、4、6、8週處死大鼠，觀察肝組織炎癥及纖維化程度，放射免疫法檢測血清中透明質痠（HA）的含量，透射電鏡觀察肝竇竇壁結構，S-P 免疫組織化學檢測各組大鼠肝組織CD31、層黏連蛋白（LN）、IV型膠原（Col-IV）的錶達。結果肝髒組織學證實CCl4誘導的大鼠肝纖維化模型構建成功，6週可見纖維間隔形成；透射電鏡顯示，CCl4誘導2週時，部分肝竇內皮細胞（liver sinusoidal endothelial cells ，LSEC）窗孔減少，內皮下未見基底膜（Basement membrane，BM），隨著造模的進程，LSEC 窗孔進一步減少，部分甚至消失，第6、8週時跼部肝竇內皮下形成連續的BM。同時，隨著肝纖維化的進程，HA濃度逐漸升高，肝竇內皮細胞錶麵標誌物CD31及基底膜主要成分Col-IV、LN錶達逐漸增彊。結論在CCl4誘導大鼠肝纖維化過程中，肝竇毛細血管化是逐漸形成的，LSEC失窗孔早于纖維間隔的形成，而肝竇內皮下基底膜齣現在纖維間隔形成以後。
목적：관찰사록화탄（CCl4）유도적대서간섬유화과정중간두모세혈관화적형성과정，탐토기여간섬유화적관계。방법32지청길급웅성SD대서，수궤분위정상대조조，간섬유화모형조，정상대조조대서복강주사0．9％록화납용액2 mL/kg ，모형조대서복강주사50％ CCl4-비마유혼합액2 mL/kg ，매주2차，공8주；분별우조모제2、4、6、8주처사대서，관찰간조직염증급섬유화정도，방사면역법검측혈청중투명질산（HA）적함량，투사전경관찰간두두벽결구，S-P 면역조직화학검측각조대서간조직CD31、층점련단백（LN）、IV형효원（Col-IV）적표체。결과간장조직학증실CCl4유도적대서간섬유화모형구건성공，6주가견섬유간격형성；투사전경현시，CCl4유도2주시，부분간두내피세포（liver sinusoidal endothelial cells ，LSEC）창공감소，내피하미견기저막（Basement membrane，BM），수착조모적진정，LSEC 창공진일보감소，부분심지소실，제6、8주시국부간두내피하형성련속적BM。동시，수착간섬유화적진정，HA농도축점승고，간두내피세포표면표지물CD31급기저막주요성분Col-IV、LN표체축점증강。결론재CCl4유도대서간섬유화과정중，간두모세혈관화시축점형성적，LSEC실창공조우섬유간격적형성，이간두내피하기저막출현재섬유간격형성이후。
Objective To observe the formation of sinusoidal capillarization in the process of liver fibrosis induced by carbon tetrachloride (CCl4 )in rats,and to explore the relevance between sinusoidal capillarization and hepatic fibrosis. Methods Thirty-two SD rats were divided randomly into control group (group N)and fibrosis model group (group M). Rats in group N were treated with 2 ml/kg normal sodium twice a week for eight weeks via intraperitoneal injection,while rats in group M were treated with 2 ml/kg a mixture of 50% CCl4 and castor oil twice a week for eight weeks. Rats were executed at the end of 2th wk,4th wk,6th wk,8th wk to collect samples of liver tissue and serum. Grading and staging of hepatic fibrosis were measured by HE staining and masson staining. Changes of HA from serum were measured by radioimmunoassay. Structure of sinus wall in liver sinusoids was observed by TEM. Expressions of CD3 1 ,LN and Col-IV in liver tissue were measured by S-P immunohistochemistry. Results Liver pathohistology analysis confirmed that hepatic fibrosis rat model induced by CCl4 was established successfully. Fibrous septum could be observed at the end of 6th wk. TEM showed that fenestrae of some liver sinusoidal endothelial cells (LSECs)diminished after treated with CCl4 for two weeks with observing no basement membrane (BM). Fenestrae of LSECs diminished along with the development of hepatic fibrosis,even disappeared. Consecutive BM formed at the end of 6th wk~8th wk in local sinusoidal endothelium. Concentration of HA from serum,and expressions of CD31 ,Col-IV and LN in hepatic sinusoids increased gradually along with development of hepatic fibrosis. Conclusion Formation of sinusoidal capillarization was occurred gradually along with development of hepatic fibrosis. Fibrous septa formed later than fenestrae losing in LSECs,but earlier than formation of sinusoidal endothelial basement membrane.
