重庆医学
重慶醫學
중경의학
CHONGQING MEDICAL JOURNAL
2015年
6期
724-726,729
,共4页
彭科%许超%于敏%孙力华%肖卫东%杨桦
彭科%許超%于敏%孫力華%肖衛東%楊樺
팽과%허초%우민%손력화%초위동%양화
S-亚硝基谷胱甘肽%肠黏膜屏障%紧密连接蛋白%肠缺血再灌注
S-亞硝基穀胱甘肽%腸黏膜屏障%緊密連接蛋白%腸缺血再灌註
S-아초기곡광감태%장점막병장%긴밀련접단백%장결혈재관주
S-Nitrosoglutathione%intestinal epithelial barrier%tight junction protein%intestinal ischemia reperfusion
目的:探讨 S-亚硝基谷胱甘肽(GSNO)预处理对小鼠肠急性缺血再灌注(I/R)所致的肠黏膜屏障损伤的影响。方法6~8周龄雄性 C57BL/6小鼠24只,分为 Sham 组、I/R 组、I/R+GSNO 组,每组8只。采用夹闭肠系膜上动脉30 min,再灌注6 h作为小鼠肠 I/R 模型。苏木素-伊红(HE)染色观察小肠组织形态学的变化;免疫组织化学观察肠上皮细胞间紧密连接蛋白 clau-din-1的表达情况;Western blot 检测 claudin-1蛋白水平变化。结果 HE 染色显示,与 Sham 组比较,I/R 组肠黏膜明显肿胀,绒毛部分脱落、变粗,绒毛倒伏、断裂,而 I/R+GSNO 组肠黏膜无明显肿胀,绒毛完整,形态接近正常;免疫组织化学观察显示 I/R刺激可造成肠上皮表面 claudin-1连续性明显破坏,阳性表达率明显降低,I/R+GSNO 组肠上皮 claudin-1连续性明显恢复,阳性染色显著增加;肠上皮 claudin-1蛋白定量分析显示:与 Sham 组比较,I/R 组及 I/R+GSNO 小肠上皮紧密连接蛋白 claudin-1表达分别下降32.5%,13.8%(P <0.05);与 I/R 组比较,I/R+GSNO 组小鼠肠上皮紧密连接蛋白 claudin-1表达上调27.8%(P <0.05)。结论小肠急性 I/R 刺激可造成明显的肠黏膜损伤及肠上皮间紧密连接的显著破坏;GSNO 预处理可通过上调 claudin-1表达,有效缓解 I/R 对肠黏膜结构与肠上皮屏障功能的损伤。
目的:探討 S-亞硝基穀胱甘肽(GSNO)預處理對小鼠腸急性缺血再灌註(I/R)所緻的腸黏膜屏障損傷的影響。方法6~8週齡雄性 C57BL/6小鼠24隻,分為 Sham 組、I/R 組、I/R+GSNO 組,每組8隻。採用夾閉腸繫膜上動脈30 min,再灌註6 h作為小鼠腸 I/R 模型。囌木素-伊紅(HE)染色觀察小腸組織形態學的變化;免疫組織化學觀察腸上皮細胞間緊密連接蛋白 clau-din-1的錶達情況;Western blot 檢測 claudin-1蛋白水平變化。結果 HE 染色顯示,與 Sham 組比較,I/R 組腸黏膜明顯腫脹,絨毛部分脫落、變粗,絨毛倒伏、斷裂,而 I/R+GSNO 組腸黏膜無明顯腫脹,絨毛完整,形態接近正常;免疫組織化學觀察顯示 I/R刺激可造成腸上皮錶麵 claudin-1連續性明顯破壞,暘性錶達率明顯降低,I/R+GSNO 組腸上皮 claudin-1連續性明顯恢複,暘性染色顯著增加;腸上皮 claudin-1蛋白定量分析顯示:與 Sham 組比較,I/R 組及 I/R+GSNO 小腸上皮緊密連接蛋白 claudin-1錶達分彆下降32.5%,13.8%(P <0.05);與 I/R 組比較,I/R+GSNO 組小鼠腸上皮緊密連接蛋白 claudin-1錶達上調27.8%(P <0.05)。結論小腸急性 I/R 刺激可造成明顯的腸黏膜損傷及腸上皮間緊密連接的顯著破壞;GSNO 預處理可通過上調 claudin-1錶達,有效緩解 I/R 對腸黏膜結構與腸上皮屏障功能的損傷。
목적:탐토 S-아초기곡광감태(GSNO)예처리대소서장급성결혈재관주(I/R)소치적장점막병장손상적영향。방법6~8주령웅성 C57BL/6소서24지,분위 Sham 조、I/R 조、I/R+GSNO 조,매조8지。채용협폐장계막상동맥30 min,재관주6 h작위소서장 I/R 모형。소목소-이홍(HE)염색관찰소장조직형태학적변화;면역조직화학관찰장상피세포간긴밀련접단백 clau-din-1적표체정황;Western blot 검측 claudin-1단백수평변화。결과 HE 염색현시,여 Sham 조비교,I/R 조장점막명현종창,융모부분탈락、변조,융모도복、단렬,이 I/R+GSNO 조장점막무명현종창,융모완정,형태접근정상;면역조직화학관찰현시 I/R자격가조성장상피표면 claudin-1련속성명현파배,양성표체솔명현강저,I/R+GSNO 조장상피 claudin-1련속성명현회복,양성염색현저증가;장상피 claudin-1단백정량분석현시:여 Sham 조비교,I/R 조급 I/R+GSNO 소장상피긴밀련접단백 claudin-1표체분별하강32.5%,13.8%(P <0.05);여 I/R 조비교,I/R+GSNO 조소서장상피긴밀련접단백 claudin-1표체상조27.8%(P <0.05)。결론소장급성 I/R 자격가조성명현적장점막손상급장상피간긴밀련접적현저파배;GSNO 예처리가통과상조 claudin-1표체,유효완해 I/R 대장점막결구여장상피병장공능적손상。
Objective To investigate the effect of S-Nitrosoglutathione (GSNO)on acute ischemia reperfusion (I/R)induced in-testinal barrier function lesion in a mouse model.Methods Twenty-four 6-8-year-old C57BL/6 mice were divided into 3 groups,8 for each:(1)the sham group;(2)the I/R group;(3)the I/R+GSNO group.The mouse intestine I/R model was established by the occlusion of the superior mesenteric artery temporarily followed by reperfusion for 6 h.Histological changes in the small intestine were observed after HE staining;the expression of claudin-1 protein in the intestine epithelium was assessed by immunohistochem-istry staining as well as western blot analysis.Results Both HE staining and immunohistochemistry results showed the integrate intestinal villi with the continuous Claudin-1 expression alone the villi in the sham group;the intestinal villi of the I/R group partial-ly detached,thickened,crooked and fractured,with the obvious disconnection of Claudin-1 staining alone the top of the villi;while the intestinal villi of the I/R+GSNO group were neatly arranged and damage to intestinal mucosa was much alleviated,accompanied with the marked restoration of the continuity of claudin-1 staining.Compared to the sham group,claudin-1 protein level of for the I/R group and the I/R+GSNO group decreased by 32.5% and 13.8% respectively (P <0.05);and compared to the I/R group,clau-din-1 protein level of the I/R+GSNO group increased by 27.8% (P <0.05).Conclusion Protein level of claudin-1 would decrea-ses after I/R,and pretreatment with GSNO can effectively relieve the damage of intestinal mucosal structure as well as intestinal tight junction barrier through upregulating the expression of claudin-1 protein.