中华围产医学杂志
中華圍產醫學雜誌
중화위산의학잡지
CHINESE JOURNAL OF PERINATAL MEDICINE
2011年
12期
739-743
,共5页
薛春红%曾万江%乔福元%刘海意%祝秀
薛春紅%曾萬江%喬福元%劉海意%祝秀
설춘홍%증만강%교복원%류해의%축수
肺泡%肺表面活性物质相关蛋白质B%转化生长因子β1%血管内皮生长因子A%地塞米松
肺泡%肺錶麵活性物質相關蛋白質B%轉化生長因子β1%血管內皮生長因子A%地塞米鬆
폐포%폐표면활성물질상관단백질B%전화생장인자β1%혈관내피생장인자A%지새미송
Pulmonary alveoli%Pulmonary surfactant-associated protein B%Transforming growth factor beta 1%Vascular endothelial growth factor A%Daxamethasone
目的 探讨血管内皮生长因子(vascular endothelial growth factor,VEGF)及地塞米松对肺泡Ⅱ型上皮细胞(typeⅡalveolar epithelial cell,AECⅡ)表面活性蛋白-B(surfactant protein B,SP-B)和转化生长因子-β1(transforming growth factor-β1,TGF-β1)表达的影响及意义. 方法 原代培养早产鼠AECⅡ,采用实时荧光定量聚合酶链反应技术(real-time quantitative polymerasechain reaction,real-time PCR)、免疫细胞化学染色等方法,观察不同剂量的VEGF(25 ng/ml组、50 ng/ml组和100 ng/ml组)和不同剂量的地塞米松(25 nmol/ml组、50 nmol/ml组、100 nmol/ml组、200 nmol/ml组)对AECⅡSP-B mRNA及TGF-β1 mRNA表达的影响.同时设一对照组.采用方差分析或q检验进行统计学分析. 结果 与对照组相比,25 ng/ml VEGF组及25 nmol/ml、50 nmol/ml、100 nmol/ml和200 nmol/ml地塞米松各组SP-B mRNA表达水平明显上升(13.500±3.172、3.547±0.690、5.219±0.782、3.493±0.335和3.981±1.133与1.001±0.059,q分别为-5.286、-4.943、-7.228、-9.906、-3.525,P均＜0.05).25 ng/ml VEGF组及50、100、200 nmol/ml地塞米松组TGF-β1 mRNA表达明显下降(0.451±0.078、0.579±0.019、0.422±0.020和0.769±0.025与1.019±0.226,q分别为4.110、3.356、4.551、1.901,P均＜0.05),其他各组与对照组相比差异无统计学意义(P＞0.05).免疫细胞化学结果显示,25 ng/ml VEGF组及50、100、200 nmol/ml地塞米松组AECⅡ染色阳性率分别为23％、26％、22％和29％；对照组AECⅡ染色阳性率为80％.结论 VEGF及地塞米松均有促进SP-B表达的作用,TGF-β1可能对SP-B的表达有负调控作用,VEGF和地塞米松可能是通过调控与TGF-β1有关的调控通路发挥促进SP-B表达的作用.
目的 探討血管內皮生長因子(vascular endothelial growth factor,VEGF)及地塞米鬆對肺泡Ⅱ型上皮細胞(typeⅡalveolar epithelial cell,AECⅡ)錶麵活性蛋白-B(surfactant protein B,SP-B)和轉化生長因子-β1(transforming growth factor-β1,TGF-β1)錶達的影響及意義. 方法 原代培養早產鼠AECⅡ,採用實時熒光定量聚閤酶鏈反應技術(real-time quantitative polymerasechain reaction,real-time PCR)、免疫細胞化學染色等方法,觀察不同劑量的VEGF(25 ng/ml組、50 ng/ml組和100 ng/ml組)和不同劑量的地塞米鬆(25 nmol/ml組、50 nmol/ml組、100 nmol/ml組、200 nmol/ml組)對AECⅡSP-B mRNA及TGF-β1 mRNA錶達的影響.同時設一對照組.採用方差分析或q檢驗進行統計學分析. 結果 與對照組相比,25 ng/ml VEGF組及25 nmol/ml、50 nmol/ml、100 nmol/ml和200 nmol/ml地塞米鬆各組SP-B mRNA錶達水平明顯上升(13.500±3.172、3.547±0.690、5.219±0.782、3.493±0.335和3.981±1.133與1.001±0.059,q分彆為-5.286、-4.943、-7.228、-9.906、-3.525,P均＜0.05).25 ng/ml VEGF組及50、100、200 nmol/ml地塞米鬆組TGF-β1 mRNA錶達明顯下降(0.451±0.078、0.579±0.019、0.422±0.020和0.769±0.025與1.019±0.226,q分彆為4.110、3.356、4.551、1.901,P均＜0.05),其他各組與對照組相比差異無統計學意義(P＞0.05).免疫細胞化學結果顯示,25 ng/ml VEGF組及50、100、200 nmol/ml地塞米鬆組AECⅡ染色暘性率分彆為23％、26％、22％和29％；對照組AECⅡ染色暘性率為80％.結論 VEGF及地塞米鬆均有促進SP-B錶達的作用,TGF-β1可能對SP-B的錶達有負調控作用,VEGF和地塞米鬆可能是通過調控與TGF-β1有關的調控通路髮揮促進SP-B錶達的作用.
목적 탐토혈관내피생장인자(vascular endothelial growth factor,VEGF)급지새미송대폐포Ⅱ형상피세포(typeⅡalveolar epithelial cell,AECⅡ)표면활성단백-B(surfactant protein B,SP-B)화전화생장인자-β1(transforming growth factor-β1,TGF-β1)표체적영향급의의. 방법 원대배양조산서AECⅡ,채용실시형광정량취합매련반응기술(real-time quantitative polymerasechain reaction,real-time PCR)、면역세포화학염색등방법,관찰불동제량적VEGF(25 ng/ml조、50 ng/ml조화100 ng/ml조)화불동제량적지새미송(25 nmol/ml조、50 nmol/ml조、100 nmol/ml조、200 nmol/ml조)대AECⅡSP-B mRNA급TGF-β1 mRNA표체적영향.동시설일대조조.채용방차분석혹q검험진행통계학분석. 결과 여대조조상비,25 ng/ml VEGF조급25 nmol/ml、50 nmol/ml、100 nmol/ml화200 nmol/ml지새미송각조SP-B mRNA표체수평명현상승(13.500±3.172、3.547±0.690、5.219±0.782、3.493±0.335화3.981±1.133여1.001±0.059,q분별위-5.286、-4.943、-7.228、-9.906、-3.525,P균＜0.05).25 ng/ml VEGF조급50、100、200 nmol/ml지새미송조TGF-β1 mRNA표체명현하강(0.451±0.078、0.579±0.019、0.422±0.020화0.769±0.025여1.019±0.226,q분별위4.110、3.356、4.551、1.901,P균＜0.05),기타각조여대조조상비차이무통계학의의(P＞0.05).면역세포화학결과현시,25 ng/ml VEGF조급50、100、200 nmol/ml지새미송조AECⅡ염색양성솔분별위23％、26％、22％화29％；대조조AECⅡ염색양성솔위80％.결론 VEGF급지새미송균유촉진SP-B표체적작용,TGF-β1가능대SP-B적표체유부조공작용,VEGF화지새미송가능시통과조공여TGF-β1유관적조공통로발휘촉진SP-B표체적작용.
Objective To investigate the effects of vascular endothelial growth factor (VEGF) and dexamethasone on mRNA expressions of surfactant protein B (SP-B) and transforming growth factor-β1 (TGF-β1) of type Ⅱ alveolar epithelial cell (AECⅡ). Methods AECⅡ were isolated and purified from fetal rat lung tissues,then cultured with different dose of VEGF (25,50 and 100 ng/ml) and dexamethasone (25,50,100 and 200 nmol/ml).The mRNA levels of SP-B and TGF-β1 were detected by real-time quantitative polymerase chain reaction (RT-PCR) and expression of TGF-β1 protein was detected by immunocytochemistry.ANOVA or q-test was applied to compare the difference among groups.Results Compared with control group,SP-B mRNA levels in 25 ng/ml VEGF group and 25,50,100 and 200 nmol/ml dexamethasone groups were higher (13.500±3.172,3.547±0.690,5.219±0.782,3.493±0.335,and 3.981 ± 1.133 vs 1.001 ± 0.059,q=-5.286,-4.943,- 7.228,- 9.906 and - 3.525 respectively,P＜0.05) ; TGF-β1 mRNA expression of 25 ng/ml VEGF group,50,100 and 200 nmol/ml dexamethasone group was lower (0.451 ± 0.078,0.579±0.019,0.422 ± 0.020 and 0.769 ± 0.025 vs 1.019±0.226,q=4.110,3.356,4.551 and 1.901 respectively,P＜0.05) ; other groups had no significant differences compared with control group (P＞0.05).Immunocytochemistry showed that the positive rate of TGF-β1 expression in 25 ng/ml VEGF,50,100 and 200 nmol/ml dexamethasone group was 23％,26％,22％ and 29％,respectively,while in the control group,the expression of TGF-β1 was positive in most of the AECⅡ (80％).Conclusions Both VEGF and dexamethasone could increase the expression of SP-B at mRNA level at appropriate concentrations.At the same time,the expression of TGF-β1 is inhibited.It is suggested that both VEGF and dexamethasone might increase the mRNA expression of SP-B by inhibiting the expression of TGF-β1.
