解剖学报
解剖學報
해부학보
ACTA ANATOMICA SINICA
2014年
5期
704-709
,共6页
宁巍%廖晓岗%王毅%姚志勇%毛胜艳
寧巍%廖曉崗%王毅%姚誌勇%毛勝豔
저외%료효강%왕의%요지용%모성염
镉%黄芪甲苷%闭锁小带-1蛋白%紧密连接蛋白-11%P-p38 MAPK%免疫组织化学%免疫印迹法%大鼠
鎘%黃芪甲苷%閉鎖小帶-1蛋白%緊密連接蛋白-11%P-p38 MAPK%免疫組織化學%免疫印跡法%大鼠
력%황기갑감%폐쇄소대-1단백%긴밀련접단백-11%P-p38 MAPK%면역조직화학%면역인적법%대서
Cadmium%Astragaloside IV%Zonula occludens-1%Claudin-11%P-p38 MAPK%Immunohistochemistry%Western blotting%Rat
目的：观察黄芪甲苷和SB203580对镉致大鼠血睾屏障破坏及相关蛋白表达改变的保护效应，探讨黄芪甲苷的保护机制。方法21只成年雄性SD大鼠随机分为7组：单纯镉组[0．1％氯化镉腹腔内注射，1mg／（kg?d）]，镉＋黄芪甲苷组[镉剂量同上，同时注射黄芪甲苷，10mg／（kg?d）]，镉＋SB203580组[镉剂量同上，同时注射SB203580，100μg／（kg?d）]，以上各组又分为连续处理5d和10d两个时间组，对照组腹腔内注射等量生理盐水。各实验和对照组动物均为3只。取睾丸做光学显微镜、电子显微镜观察以及免疫组织化学染色和Western blotting检测。结果 HE染色观察对照组支持细胞核染色较浅且不规则，镉组支持细胞内有空泡形成，镉＋黄芪甲苷组与镉＋SB203580组未见明显形态异常。免疫组织化学染色观察对照组闭锁小带-1蛋白（ ZO-1）、紧密连接蛋白-11（ claudin-11）阳性产物在生精上皮靠近基底部表达。镉组ZO-1、claudin-11阳性产物表达均显著降低（ P＜0．05），镉＋黄芪甲苷组与镉＋SB203580组阳性产物表达低于对照组但明显高于镉组（P＜0．05）。超微结构观察对照组血睾屏障紧密连接形态完整，呈连续的电子密度较深致密线，镉组血睾屏障紧密连接及支持细胞均见不同程度破坏，镉＋黄芪甲苷组与镉＋SB203580组破坏程度较相应处理时间镉组为轻。 Western blotting 结果显示，镉组磷酸化p38丝裂原活化蛋白激酶（p-p38MAPK）水平明显增强（P＜0．05），镉＋黄芪甲苷组与镉＋SB203580组p-p38MAPK水平虽高于对照组，但较镉组明显减弱（ P＜0．05）。结论镉致大鼠血睾屏障ZO-1、claudin-11表达降低，紧密连接超微结构损伤，黄芪甲苷具有保护作用，其保护机制与抑制p38MAPK磷酸化有关。
目的：觀察黃芪甲苷和SB203580對鎘緻大鼠血睪屏障破壞及相關蛋白錶達改變的保護效應，探討黃芪甲苷的保護機製。方法21隻成年雄性SD大鼠隨機分為7組：單純鎘組[0．1％氯化鎘腹腔內註射，1mg／（kg?d）]，鎘＋黃芪甲苷組[鎘劑量同上，同時註射黃芪甲苷，10mg／（kg?d）]，鎘＋SB203580組[鎘劑量同上，同時註射SB203580，100μg／（kg?d）]，以上各組又分為連續處理5d和10d兩箇時間組，對照組腹腔內註射等量生理鹽水。各實驗和對照組動物均為3隻。取睪汍做光學顯微鏡、電子顯微鏡觀察以及免疫組織化學染色和Western blotting檢測。結果 HE染色觀察對照組支持細胞覈染色較淺且不規則，鎘組支持細胞內有空泡形成，鎘＋黃芪甲苷組與鎘＋SB203580組未見明顯形態異常。免疫組織化學染色觀察對照組閉鎖小帶-1蛋白（ ZO-1）、緊密連接蛋白-11（ claudin-11）暘性產物在生精上皮靠近基底部錶達。鎘組ZO-1、claudin-11暘性產物錶達均顯著降低（ P＜0．05），鎘＋黃芪甲苷組與鎘＋SB203580組暘性產物錶達低于對照組但明顯高于鎘組（P＜0．05）。超微結構觀察對照組血睪屏障緊密連接形態完整，呈連續的電子密度較深緻密線，鎘組血睪屏障緊密連接及支持細胞均見不同程度破壞，鎘＋黃芪甲苷組與鎘＋SB203580組破壞程度較相應處理時間鎘組為輕。 Western blotting 結果顯示，鎘組燐痠化p38絲裂原活化蛋白激酶（p-p38MAPK）水平明顯增彊（P＜0．05），鎘＋黃芪甲苷組與鎘＋SB203580組p-p38MAPK水平雖高于對照組，但較鎘組明顯減弱（ P＜0．05）。結論鎘緻大鼠血睪屏障ZO-1、claudin-11錶達降低，緊密連接超微結構損傷，黃芪甲苷具有保護作用，其保護機製與抑製p38MAPK燐痠化有關。
목적：관찰황기갑감화SB203580대력치대서혈고병장파배급상관단백표체개변적보호효응，탐토황기갑감적보호궤제。방법21지성년웅성SD대서수궤분위7조：단순력조[0．1％록화력복강내주사，1mg／（kg?d）]，력＋황기갑감조[력제량동상，동시주사황기갑감，10mg／（kg?d）]，력＋SB203580조[력제량동상，동시주사SB203580，100μg／（kg?d）]，이상각조우분위련속처리5d화10d량개시간조，대조조복강내주사등량생리염수。각실험화대조조동물균위3지。취고환주광학현미경、전자현미경관찰이급면역조직화학염색화Western blotting검측。결과 HE염색관찰대조조지지세포핵염색교천차불규칙，력조지지세포내유공포형성，력＋황기갑감조여력＋SB203580조미견명현형태이상。면역조직화학염색관찰대조조폐쇄소대-1단백（ ZO-1）、긴밀련접단백-11（ claudin-11）양성산물재생정상피고근기저부표체。력조ZO-1、claudin-11양성산물표체균현저강저（ P＜0．05），력＋황기갑감조여력＋SB203580조양성산물표체저우대조조단명현고우력조（P＜0．05）。초미결구관찰대조조혈고병장긴밀련접형태완정，정련속적전자밀도교심치밀선，력조혈고병장긴밀련접급지지세포균견불동정도파배，력＋황기갑감조여력＋SB203580조파배정도교상응처리시간력조위경。 Western blotting 결과현시，력조린산화p38사렬원활화단백격매（p-p38MAPK）수평명현증강（P＜0．05），력＋황기갑감조여력＋SB203580조p-p38MAPK수평수고우대조조，단교력조명현감약（ P＜0．05）。결론력치대서혈고병장ZO-1、claudin-11표체강저，긴밀련접초미결구손상，황기갑감구유보호작용，기보호궤제여억제p38MAPK린산화유관。
Objective To observe the effect of astragaloside IV (A) and SB203580 antagonize cadmium (Cd) toxicity on expression of associated protein and blood-testis barrier(BTB) in rats and to study the protective mechanism of A on it.Methods Totally 21 SD male rats were randomly divided into 7 groups, 3 rats per group:Cd [ intraperitoneally injected with 0.1%CdCl2,1mg/(kg?d)],Cd+A [at the above dose of CdCl2,at the same time with A,10mg/(kg?d)], and Cd +SB203580 [at the above dose of CdCl2,at the same time with SB203580,100μg/(kg?d)], each of the above groups was further divided into continuous five and ten days treatment groups .The control group was intraperitoneally injected with equal dosage of normal saline .The testes were studied by light , electron microscopy , immunohistochemistry and Western blotting .Results In the control group ,irregular and lightly stained nuclei of Sertoli cell ( Sc) in seminiferous tubules were observed by HE staining .A continuous electron density line of tight junction ( TJ) and normal ultrastructure of BTB were observed .After Cd treatment ,the vesicular formation in the Sc was observed .The ultrastructural damage of Sc and TJ was observed .Compared with the corresponding time point of Cd group ,these were weakened in morphology of testis and ultrastructure of TJ after Cd +A or Cd +SB203580 treatment .The positive products of zonula occludens-1 ( ZO-1 ) and claudin-11 were localized mainly in the base of the seminiferous tubule .After Cd treatment , the average absorbance (AA) of ZO-1 and Claudin-11 was decreased significantly compared with that of the control group (P<0.05).After Cd +A or Cd +SB203580 treatment,AA of ZO-1 and Claudin-11 were increased significantly compared with that of the Cd group(P<0.05),though lower than that of the control group .The result of Western blotting showed that phosphorylation-p38MAPK in Cd group was increased significantly compared with that of the control group (P<0.05).After Cd +A or Cd+SB203580 treatment, it was decreased significantly compared with that of the Cd group (P<0.05).Conclusion Cd decreases ZO-1 and Claudin-11 expression and damages ultrastructure of TJ in BTB , asⅣhas protective effect on it , and is related to inhibiting activation of p 38 MAPK pathway .
