中华急诊医学杂志
中華急診醫學雜誌
중화급진의학잡지
CHINESE JOURNAL OF EMERGENCY MEDICINE
2009年
5期
457-461
,共5页
刘斌剑%祁朝玲%郑淑辉%周木秀%王建民
劉斌劍%祁朝玲%鄭淑輝%週木秀%王建民
류빈검%기조령%정숙휘%주목수%왕건민
角质细胞生长因子受体%钠离子通道%急性肺损伤%基因治疗
角質細胞生長因子受體%鈉離子通道%急性肺損傷%基因治療
각질세포생장인자수체%납리자통도%급성폐손상%기인치료
Keratinocyte growth factor receptor%Sodium ehannol%Acute lung injury%Gem therapy
目的 观察角质细胞生长因子受体(KGFR)腺病毒表达载体在大鼠急性肺损伤前后对肺泡Ⅱ型上皮细胞钠离子通道表达的促进作用,从而验证基因治疗急性肺损伤的效果.方法 将雄性SD大鼠40只分成4组:正常对照组(n=8),致伤对照组(n=10),正常转基因组(n=10),致伤转基因组(n=12).用脂多糖(LPS)造成急性肺损伤模型,以肺组织明显肿大病变为制模成功标准.正常转基因组和致伤转基因组均通过大鼠尾静脉注射KGFR基因的腺病毒表达载体应用液1 mL,72 h后致伤转基因组和致伤对照组分别通过大鼠尾静脉以5 mg/kg注射LPS,正常转基因组和正常对照组则注射等量的生理盐水.过48 h后,断头处死所有实验组大鼠,取肺组织进行实验.通过免疫组化和免疫电镜检测各组大鼠肺泡Ⅱ型上皮细胞钠离子通道的表达情况.数据采用完全随机区组设计的方差分析,行LSD-t检验,以P<0.05为差异具有统计学意义.结果 肺泡Ⅱ型上皮细胞钠离子通道表达水平(免疫组化)由高到低依次为:正常对照组(PU值=47.7±3.33),正常转基因组(PU值=46.9±5.21),致伤转基因组(PU值=29.19±4.11)和致伤对照组(PU值=5.1±2.3);致伤转基因组的钠离子通道表达水平低于正常转基因组(t=9.134,P<0.001)和正常对照组(t=10.601,P<0.001),但明显高于致伤对照组(t=16.466,P<0.001).结论 KGFR腺病毒表达载体转基因效果明显,能够在LPS致伤情况下有效促进钠离子通道的表达,缓解肺泡腔内钠水潴留,达到转基因治疗肺损伤的效果.
目的 觀察角質細胞生長因子受體(KGFR)腺病毒錶達載體在大鼠急性肺損傷前後對肺泡Ⅱ型上皮細胞鈉離子通道錶達的促進作用,從而驗證基因治療急性肺損傷的效果.方法 將雄性SD大鼠40隻分成4組:正常對照組(n=8),緻傷對照組(n=10),正常轉基因組(n=10),緻傷轉基因組(n=12).用脂多糖(LPS)造成急性肺損傷模型,以肺組織明顯腫大病變為製模成功標準.正常轉基因組和緻傷轉基因組均通過大鼠尾靜脈註射KGFR基因的腺病毒錶達載體應用液1 mL,72 h後緻傷轉基因組和緻傷對照組分彆通過大鼠尾靜脈以5 mg/kg註射LPS,正常轉基因組和正常對照組則註射等量的生理鹽水.過48 h後,斷頭處死所有實驗組大鼠,取肺組織進行實驗.通過免疫組化和免疫電鏡檢測各組大鼠肺泡Ⅱ型上皮細胞鈉離子通道的錶達情況.數據採用完全隨機區組設計的方差分析,行LSD-t檢驗,以P<0.05為差異具有統計學意義.結果 肺泡Ⅱ型上皮細胞鈉離子通道錶達水平(免疫組化)由高到低依次為:正常對照組(PU值=47.7±3.33),正常轉基因組(PU值=46.9±5.21),緻傷轉基因組(PU值=29.19±4.11)和緻傷對照組(PU值=5.1±2.3);緻傷轉基因組的鈉離子通道錶達水平低于正常轉基因組(t=9.134,P<0.001)和正常對照組(t=10.601,P<0.001),但明顯高于緻傷對照組(t=16.466,P<0.001).結論 KGFR腺病毒錶達載體轉基因效果明顯,能夠在LPS緻傷情況下有效促進鈉離子通道的錶達,緩解肺泡腔內鈉水潴留,達到轉基因治療肺損傷的效果.
목적 관찰각질세포생장인자수체(KGFR)선병독표체재체재대서급성폐손상전후대폐포Ⅱ형상피세포납리자통도표체적촉진작용,종이험증기인치료급성폐손상적효과.방법 장웅성SD대서40지분성4조:정상대조조(n=8),치상대조조(n=10),정상전기인조(n=10),치상전기인조(n=12).용지다당(LPS)조성급성폐손상모형,이폐조직명현종대병변위제모성공표준.정상전기인조화치상전기인조균통과대서미정맥주사KGFR기인적선병독표체재체응용액1 mL,72 h후치상전기인조화치상대조조분별통과대서미정맥이5 mg/kg주사LPS,정상전기인조화정상대조조칙주사등량적생리염수.과48 h후,단두처사소유실험조대서,취폐조직진행실험.통과면역조화화면역전경검측각조대서폐포Ⅱ형상피세포납리자통도적표체정황.수거채용완전수궤구조설계적방차분석,행LSD-t검험,이P<0.05위차이구유통계학의의.결과 폐포Ⅱ형상피세포납리자통도표체수평(면역조화)유고도저의차위:정상대조조(PU치=47.7±3.33),정상전기인조(PU치=46.9±5.21),치상전기인조(PU치=29.19±4.11)화치상대조조(PU치=5.1±2.3);치상전기인조적납리자통도표체수평저우정상전기인조(t=9.134,P<0.001)화정상대조조(t=10.601,P<0.001),단명현고우치상대조조(t=16.466,P<0.001).결론 KGFR선병독표체재체전기인효과명현,능구재LPS치상정황하유효촉진납리자통도적표체,완해폐포강내납수저류,체도전기인치료폐손상적효과.
Objective To explore the effects of keratinocyte growth factor receptor (KGFR) transgene on sodium channel in alveolar type Ⅱ cells with LPS-induced acute lung injury, to provide the evidence for gene treatment in acute lung injury. Method Totally 40 male Sprague-Dawtey rats were randomly divided into four groups, including normal control (n=8), injured control (n=10), normal transgene (n=10) and injured transgene (n=12). The models of acute lung injury were produced using LPS, and the successful criteria was the obvious enlargement in the lung tissue. The rats in normal transgene group and injured transgene group were injected with 1 mL of KGFR adenovirus vector through rats' tail vein. At 72 hours later, the rats in injured control group and injured transgene group were injected with LPS in dose of 5 mg/kg (BW). While rats in normal control group and normal transgene group were injected with equivalent saline simultaneously. Another 48 hours later, rats in the four groups were killed. The lung tissue were collected for analysis. The expression of sodium channel in rats' alveolar type Ⅱ cells were detected by immunohistochemistry and immunoeectron microscope. Difference among the experimental groups were estimated by ANOVA analysis (LSD-t-test). There was statistical signifi-cance when P<0.05. Results The levels of sodium channel expression in rats' alveolar type Ⅱ cells were differ-era, with normal control group (47.7±3.33), normal transgene group (46.9±5.21), injured tramgene group (29.19±4.11) and injured control group (5.1±2.3). The level of sodium channel expression in injured trans-gene group was lower than that in normal transgene group (t=9.134, P<0.001) and normal control group (t=10.601,P<0.001), but signifieantly higher than that in injured control group (t=16.466, P<0.001). Conclusions The transgene vector can effectively promote the expression of sodium channel in alveolar type Ⅱ cells in rats with LPS-indueed acute lung injury, and can alleviate sodium and water reteraion in alveolar.
