临床儿科杂志
臨床兒科雜誌
림상인과잡지
2015年
2期
179-183
,共5页
重组人促红细胞生成素%坏死性小肠结肠炎%肠道损伤%Bcl-2%caspase-3
重組人促紅細胞生成素%壞死性小腸結腸炎%腸道損傷%Bcl-2%caspase-3
중조인촉홍세포생성소%배사성소장결장염%장도손상%Bcl-2%caspase-3
recombinant human erythropoietin%necrotizing enterocolitis%intestinal injury%Bcl-2%caspase-3
目的：探讨重组人促红细胞生成素(rhEPO)对新生大鼠坏死性小肠结肠炎(NEC)的作用机制。方法出生48 h的新生SD大鼠60只随机分为正常对照组、NEC模型组及rhEPO干预1、2、3组。采用人工喂养、缺氧、冷刺激方法制备NEC模型；rhEPO干预组分别在鼠乳代乳品中加入rhEPO 0.1、1及10 U/ml干预3天，以苏木精-伊红(HE)染色，按Nadler标准行肠道组织病理学评分，免疫组化(二步法)检测大鼠肠道Bcl-2及活性caspase-3蛋白情况。采用图像分析系统对免疫组化的图像进行阳性表达积分光密度值测量。结果 NEC模型组大鼠造模24 h后出现腹胀、活动减少、反应迟钝，严重者出现皮肤苍白、皮温降低、呼吸节律改变，3组rhEPO干预组上述症状出现较晚且较轻。正常对照组、NEC模型组、干预1组、干预2组和干预3组新生大鼠NEC发生率分别为0%、60%、30%、18.2%和9.1%，组间差异有统计学意义(P=0.008)。5组新生大鼠肠道病理组织学评分、肠道Bcl-2表达以及活性caspase-3蛋白表达的差异均有统计学意义(P均<0.01)；NEC模型组肠道病理损伤最重，肠道活性caspase-3蛋白合成最多；予rhEPO干预后，肠道损伤有减轻趋势， Bcl-2表达上调，活性caspase-3合成减少。结论口服rhEPO能使肠道活性caspase-3蛋白合成减少，Bcl-2表达上调，降低NEC发生率，且对肠道的保护作用呈剂量依赖性。
目的：探討重組人促紅細胞生成素(rhEPO)對新生大鼠壞死性小腸結腸炎(NEC)的作用機製。方法齣生48 h的新生SD大鼠60隻隨機分為正常對照組、NEC模型組及rhEPO榦預1、2、3組。採用人工餵養、缺氧、冷刺激方法製備NEC模型；rhEPO榦預組分彆在鼠乳代乳品中加入rhEPO 0.1、1及10 U/ml榦預3天，以囌木精-伊紅(HE)染色，按Nadler標準行腸道組織病理學評分，免疫組化(二步法)檢測大鼠腸道Bcl-2及活性caspase-3蛋白情況。採用圖像分析繫統對免疫組化的圖像進行暘性錶達積分光密度值測量。結果 NEC模型組大鼠造模24 h後齣現腹脹、活動減少、反應遲鈍，嚴重者齣現皮膚蒼白、皮溫降低、呼吸節律改變，3組rhEPO榦預組上述癥狀齣現較晚且較輕。正常對照組、NEC模型組、榦預1組、榦預2組和榦預3組新生大鼠NEC髮生率分彆為0%、60%、30%、18.2%和9.1%，組間差異有統計學意義(P=0.008)。5組新生大鼠腸道病理組織學評分、腸道Bcl-2錶達以及活性caspase-3蛋白錶達的差異均有統計學意義(P均<0.01)；NEC模型組腸道病理損傷最重，腸道活性caspase-3蛋白閤成最多；予rhEPO榦預後，腸道損傷有減輕趨勢， Bcl-2錶達上調，活性caspase-3閤成減少。結論口服rhEPO能使腸道活性caspase-3蛋白閤成減少，Bcl-2錶達上調，降低NEC髮生率，且對腸道的保護作用呈劑量依賴性。
목적：탐토중조인촉홍세포생성소(rhEPO)대신생대서배사성소장결장염(NEC)적작용궤제。방법출생48 h적신생SD대서60지수궤분위정상대조조、NEC모형조급rhEPO간예1、2、3조。채용인공위양、결양、랭자격방법제비NEC모형；rhEPO간예조분별재서유대유품중가입rhEPO 0.1、1급10 U/ml간예3천，이소목정-이홍(HE)염색，안Nadler표준행장도조직병이학평분，면역조화(이보법)검측대서장도Bcl-2급활성caspase-3단백정황。채용도상분석계통대면역조화적도상진행양성표체적분광밀도치측량。결과 NEC모형조대서조모24 h후출현복창、활동감소、반응지둔，엄중자출현피부창백、피온강저、호흡절률개변，3조rhEPO간예조상술증상출현교만차교경。정상대조조、NEC모형조、간예1조、간예2조화간예3조신생대서NEC발생솔분별위0%、60%、30%、18.2%화9.1%，조간차이유통계학의의(P=0.008)。5조신생대서장도병리조직학평분、장도Bcl-2표체이급활성caspase-3단백표체적차이균유통계학의의(P균<0.01)；NEC모형조장도병리손상최중，장도활성caspase-3단백합성최다；여rhEPO간예후，장도손상유감경추세， Bcl-2표체상조，활성caspase-3합성감소。결론구복rhEPO능사장도활성caspase-3단백합성감소，Bcl-2표체상조，강저NEC발생솔，차대장도적보호작용정제량의뢰성。
Objectives To investigate the effect of rhEPO on newborn rats with necrotizing enterocolitis (NEC). Meth-ods Sixty newborn Sprague-Dewley (SD) rats at the age of 48 hours were randomly divided into 5 groups:control group, NEC group, and intervention groups 1, 2 and 3 treated with rhEPO. The rats were fed rat breast milk substitutes and stressed under hypoxia and cold exposure to establish NEC model. The rats with NEC were treated with different doses of rhEPO (0.1U/ml, 1U/ml and 10U/ml) in intervention groups. The expression of Bcl-2 and caspase-3 were measured by immunohistochemistry, and intestinal pathological changes were observed using HE staining. The value of positive expression was analyzed by IOD (integral optical density) image analysis system. Results Abdominal distention, decreased activity and unresponsiveness occurred in NEC rats 24 hours after stress exposure, and pale skin, decreased skin temperature and respiratory rhythm change were observed in severe cases. The symptoms appeared later and milder in three intervention groups. The NEC incidence of newborn rats was as followings:control group(0%), model group(60%), intervention group 1( 30%), intervention group 2(18.2%), intervention group 3(9.1%) and the difference was signiifcant between each group (P=0.008). The grades of intestinal injury, the expression of active caspase-3 and Bcl-2 were signiifcantly different among groups (P<0.01). Intestinal injury was the most severe and the expression level of active caspase-3 was the highest in NEC group. After rhEPO treatment, the intestinal injury and the production of active caspase-3 protein were decreased, and the expression of Bcl-2 was increased. Conclusions Oral rhEPO could decrease the ex-pression of intestinal active caspase 3, and increase the expression of Bcl-2. The protective effect of rhEPO on NEC is dosede-pendent.
