中华临床医师杂志（电子版）
中華臨床醫師雜誌（電子版）
중화림상의사잡지（전자판）
CHINESE JOURNAL OF CLINICIANS(ELECTRONIC VERSION)
2013年
13期
5975-5979
,共5页
任卫英%沈继平%潘刚%罗蔓%胡予
任衛英%瀋繼平%潘剛%囉蔓%鬍予
임위영%침계평%반강%라만%호여
非酒精性脂肪肝%肠黏膜屏障功能%紧密连接%ZO-1%Occludin
非酒精性脂肪肝%腸黏膜屏障功能%緊密連接%ZO-1%Occludin
비주정성지방간%장점막병장공능%긴밀련접%ZO-1%Occludin
Nonalcoholic fatty liver disease%Intestinal barrier function%Tight junction%Zonula Occludens-1%Occludin
目的：非酒精性脂肪性肝病（ NAFLD）与肠道屏障功能受损密切相关，本研究探讨非酒精性脂肪肝时肠上皮紧密连接蛋白Zonula Occluden-1（ZO-1）和Occludin的表达变化及其在肠黏膜屏障功能减退中的作用。方法30只雄性SD大鼠随机分为对照组（普通饮食）和高脂模型组，分别于造模的12周、16周检查肝指数（肝湿重/体重）、血清丙氨酸氨基转移酶（ ALT）、血清胆固醇（ TC）、甘油三酯（ TG）、肝组织匀浆丙二醛（ MDA）和超氧化物歧化酶（ SOD）水平，光镜下观察末端回肠黏膜形态，透射电镜下观察小肠上皮细胞紧密连接超微结构，免疫组化方法检测小肠紧密连接蛋白ZO-1和Occludin的表达。结果16周模型组大鼠肝脏脂肪变性明显，肝指数[（6.3±0.18）％ vs．（5.8±0.12）％]、血清 ALT [（68.61±12.12） U/L vs．（25.10±9.06）U/L]、血 TG[（1.30±0.14）mmol/L vs．（0.72±0.06）mmol/L]和肝组织匀浆 MDA 水平[（6.02±0.92）μmol/g vs．（2.15±0.66）μmol/g]均显著高于对照组（P＜0.01），肝组织匀浆SOD水平显著低于对照组[（5.12±1.88）U/g vs．（10.52±2.2）U/g，P＜0.01]；模型组大鼠末端回肠黏膜形态和小肠上皮细胞紧密连接超微结构未见明显异常，但肠上皮细胞凋亡增加。与对照组比较，16周模型组大鼠小肠ZO-1及Occludin蛋白的表达皆显著下降（0.65±0.12 vs．1.08±0.08；0.62±0.08 vs．0.95±0.10， P ＜0.01）。结论非酒精性脂肪肝时肠黏膜上皮屏障功能减退可能与小肠上皮细胞紧密连接蛋白ZO-1和Occludin的表达异常有关。
目的：非酒精性脂肪性肝病（ NAFLD）與腸道屏障功能受損密切相關，本研究探討非酒精性脂肪肝時腸上皮緊密連接蛋白Zonula Occluden-1（ZO-1）和Occludin的錶達變化及其在腸黏膜屏障功能減退中的作用。方法30隻雄性SD大鼠隨機分為對照組（普通飲食）和高脂模型組，分彆于造模的12週、16週檢查肝指數（肝濕重/體重）、血清丙氨痠氨基轉移酶（ ALT）、血清膽固醇（ TC）、甘油三酯（ TG）、肝組織勻漿丙二醛（ MDA）和超氧化物歧化酶（ SOD）水平，光鏡下觀察末耑迴腸黏膜形態，透射電鏡下觀察小腸上皮細胞緊密連接超微結構，免疫組化方法檢測小腸緊密連接蛋白ZO-1和Occludin的錶達。結果16週模型組大鼠肝髒脂肪變性明顯，肝指數[（6.3±0.18）％ vs．（5.8±0.12）％]、血清 ALT [（68.61±12.12） U/L vs．（25.10±9.06）U/L]、血 TG[（1.30±0.14）mmol/L vs．（0.72±0.06）mmol/L]和肝組織勻漿 MDA 水平[（6.02±0.92）μmol/g vs．（2.15±0.66）μmol/g]均顯著高于對照組（P＜0.01），肝組織勻漿SOD水平顯著低于對照組[（5.12±1.88）U/g vs．（10.52±2.2）U/g，P＜0.01]；模型組大鼠末耑迴腸黏膜形態和小腸上皮細胞緊密連接超微結構未見明顯異常，但腸上皮細胞凋亡增加。與對照組比較，16週模型組大鼠小腸ZO-1及Occludin蛋白的錶達皆顯著下降（0.65±0.12 vs．1.08±0.08；0.62±0.08 vs．0.95±0.10， P ＜0.01）。結論非酒精性脂肪肝時腸黏膜上皮屏障功能減退可能與小腸上皮細胞緊密連接蛋白ZO-1和Occludin的錶達異常有關。
목적：비주정성지방성간병（ NAFLD）여장도병장공능수손밀절상관，본연구탐토비주정성지방간시장상피긴밀련접단백Zonula Occluden-1（ZO-1）화Occludin적표체변화급기재장점막병장공능감퇴중적작용。방법30지웅성SD대서수궤분위대조조（보통음식）화고지모형조，분별우조모적12주、16주검사간지수（간습중/체중）、혈청병안산안기전이매（ ALT）、혈청담고순（ TC）、감유삼지（ TG）、간조직균장병이철（ MDA）화초양화물기화매（ SOD）수평，광경하관찰말단회장점막형태，투사전경하관찰소장상피세포긴밀련접초미결구，면역조화방법검측소장긴밀련접단백ZO-1화Occludin적표체。결과16주모형조대서간장지방변성명현，간지수[（6.3±0.18）％ vs．（5.8±0.12）％]、혈청 ALT [（68.61±12.12） U/L vs．（25.10±9.06）U/L]、혈 TG[（1.30±0.14）mmol/L vs．（0.72±0.06）mmol/L]화간조직균장 MDA 수평[（6.02±0.92）μmol/g vs．（2.15±0.66）μmol/g]균현저고우대조조（P＜0.01），간조직균장SOD수평현저저우대조조[（5.12±1.88）U/g vs．（10.52±2.2）U/g，P＜0.01]；모형조대서말단회장점막형태화소장상피세포긴밀련접초미결구미견명현이상，단장상피세포조망증가。여대조조비교，16주모형조대서소장ZO-1급Occludin단백적표체개현저하강（0.65±0.12 vs．1.08±0.08；0.62±0.08 vs．0.95±0.10， P ＜0.01）。결론비주정성지방간시장점막상피병장공능감퇴가능여소장상피세포긴밀련접단백ZO-1화Occludin적표체이상유관。
Objective To assess the changes of ZO-1 and Occludin protein expression in NAFLD and elucidate the mechanism of intestinal barrier injury .Methods Totally 30 male Spraque-Dawley ( SD ) rats were divided into 3 groups:Control group with normal diet ,NAFLD model group with 12 weeks high-fat diet and NAFLD model group with 16 weeks high-fat diet.The levels of serum alanine aminotransferase (ALT),total cholesterol(TC), triglyceride ( TG) and liver tissue oxygen index ( MDA and SOD ) were detected respectively .The intestinal mucosa architecture examination was examined by light microscopy .The ultra-structure of small intestine ,especially the tight junction ,was observed by transmission electron microscope .The expresses of tight junction protein ZO-1 and Occludin were detected by immunohistochemistry .Results Model rats presented with simple steatosis at the 16th week.The liver index in the model group was significantly higher than that in the control group [ ( 6.3 ±0.18 )% vs.( 5.8 ± 0.12)%]with higher levels of ALT[(68.61 ±12.12)U/L vs.(25.10 ±9.06)U/L],TG[(1.30 ±0.14)mmol/L vs.(0.72 ±0.06)mmol/L],MDA[(6.02 ±0.92)μmol/g vs.(2.15 ±0.66)μmol/g]and lower level of SOD [(5.12 ±1.88)U/g vs.(10.52 ±2.2)U/g,P<0.01].Transmission electron microscope revealed that there were few changes in the architecture of tight junction between intestinal epithelial cells .But the model animals ( both 12th week and 16th week)showed the decreased protein expression of Zonula Occludens-1(ZO-1)and Occludin in small intestine,and the changes were more obvious in the 16th week model animals (0.65 ±0.12 vs.1.08 ±0.08;0.62 ± 0.08 vs.0.95 ±0.10,P<0.01).Conclusion The small intestinal epithelial barrier function injury in NAFLD may be related to the decreased mRNA and protein expression of ZO-1 and Occludin .
