中华医学杂志
中華醫學雜誌
중화의학잡지
National Medical Journal of China
2009年
37期
2639-2643
,共5页
邓志宽%钱桂生%程赛宇%吴静
鄧誌寬%錢桂生%程賽宇%吳靜
산지관%전계생%정새우%오정
siRNA%血脑屏障%高原脑水肿%血管内皮生长因子
siRNA%血腦屏障%高原腦水腫%血管內皮生長因子
siRNA%혈뇌병장%고원뇌수종%혈관내피생장인자
siRNA%Blood Brain Barrier%High altitude cerebral edema%Vascular endothelial growth factor
目的 观察siRNA沉默脑组织血管内皮生长因子(VEGF)表达后对高原大鼠血脑屏障通透性的影响,探讨高原脑水肿的病理机制和预防方法.方法 50只Wistar大鼠随机分入正常对照组(NC)、高原对照组(HC)、脑室生理盐水对照组(SC)、siRNA静脉干预组(IVI)和siRNA脑室干预组(CVI).建立高原大鼠模型;NC组大鼠正常饲养;HC组大鼠造模前给予尾静脉内注射生理盐水;SC组在造模前给予脑室注射生理盐水;IVI组在造模前给予尾静脉注射VEGF特异性siRNA,CVI组给予脑室内注射VEGF特异性siRNA.造模24 h后迅速断头取前脑,干湿重法测定脑组织含水百分率及应用荧光素钠透过率测定血脑屏障通透性;实时荧光定量RT-PCR检测VEGF-mRNA表达,Western印迹法测定大鼠脑组织VEGF表达.结果 HC组脑组织VEGF-mRNA、VEGF表达及NaFl含量均显著高于NC组[VEGF-mRNA从(21.6±3.5)Kcopies/μg上升到(36.3 ±3.9)Kcopies/μg,VEGF从0.48±0.09增加到0.77±0.12,NaFl含量从(548±48)rfu/mg增加到(674±32)rfu/mg,均P＜0.01].与HC组相比,IVI组脑组织VEGF mRNA、VEGF表达及NaFl含量差异无统计学意义(均P＞0.05).CVI组腩组织VEGF-mRNA、VEGF表达及NaFl显著低于HC组[VEGF-mRNA从(36.3±3.9)Kcopies/μg降低到(19.9 ±4.3)Kcopies/μg,P＜0.01;VEGF从0.77±0.12降低到0.44 ±0.13,P＜0.01;NaFl含量从(674±32)rfu/mg下降到(542±77)rfu/mg,P＜0.05].各组间脑组织含水率差异无统计学意义(P＞0.05).结论 高原低氧诱发的VEGF表达增高是高原脑水肿的重要病理环节,脑室内应用特异性沉默VEGF的siRNA可阻断VEGF的表达,抑制毛细血管通透性增高,可成为预防高原脑水肿的新途径.
目的 觀察siRNA沉默腦組織血管內皮生長因子(VEGF)錶達後對高原大鼠血腦屏障通透性的影響,探討高原腦水腫的病理機製和預防方法.方法 50隻Wistar大鼠隨機分入正常對照組(NC)、高原對照組(HC)、腦室生理鹽水對照組(SC)、siRNA靜脈榦預組(IVI)和siRNA腦室榦預組(CVI).建立高原大鼠模型;NC組大鼠正常飼養;HC組大鼠造模前給予尾靜脈內註射生理鹽水;SC組在造模前給予腦室註射生理鹽水;IVI組在造模前給予尾靜脈註射VEGF特異性siRNA,CVI組給予腦室內註射VEGF特異性siRNA.造模24 h後迅速斷頭取前腦,榦濕重法測定腦組織含水百分率及應用熒光素鈉透過率測定血腦屏障通透性;實時熒光定量RT-PCR檢測VEGF-mRNA錶達,Western印跡法測定大鼠腦組織VEGF錶達.結果 HC組腦組織VEGF-mRNA、VEGF錶達及NaFl含量均顯著高于NC組[VEGF-mRNA從(21.6±3.5)Kcopies/μg上升到(36.3 ±3.9)Kcopies/μg,VEGF從0.48±0.09增加到0.77±0.12,NaFl含量從(548±48)rfu/mg增加到(674±32)rfu/mg,均P＜0.01].與HC組相比,IVI組腦組織VEGF mRNA、VEGF錶達及NaFl含量差異無統計學意義(均P＞0.05).CVI組腩組織VEGF-mRNA、VEGF錶達及NaFl顯著低于HC組[VEGF-mRNA從(36.3±3.9)Kcopies/μg降低到(19.9 ±4.3)Kcopies/μg,P＜0.01;VEGF從0.77±0.12降低到0.44 ±0.13,P＜0.01;NaFl含量從(674±32)rfu/mg下降到(542±77)rfu/mg,P＜0.05].各組間腦組織含水率差異無統計學意義(P＞0.05).結論 高原低氧誘髮的VEGF錶達增高是高原腦水腫的重要病理環節,腦室內應用特異性沉默VEGF的siRNA可阻斷VEGF的錶達,抑製毛細血管通透性增高,可成為預防高原腦水腫的新途徑.
목적 관찰siRNA침묵뇌조직혈관내피생장인자(VEGF)표체후대고원대서혈뇌병장통투성적영향,탐토고원뇌수종적병리궤제화예방방법.방법 50지Wistar대서수궤분입정상대조조(NC)、고원대조조(HC)、뇌실생리염수대조조(SC)、siRNA정맥간예조(IVI)화siRNA뇌실간예조(CVI).건립고원대서모형;NC조대서정상사양;HC조대서조모전급여미정맥내주사생리염수;SC조재조모전급여뇌실주사생리염수;IVI조재조모전급여미정맥주사VEGF특이성siRNA,CVI조급여뇌실내주사VEGF특이성siRNA.조모24 h후신속단두취전뇌,간습중법측정뇌조직함수백분솔급응용형광소납투과솔측정혈뇌병장통투성;실시형광정량RT-PCR검측VEGF-mRNA표체,Western인적법측정대서뇌조직VEGF표체.결과 HC조뇌조직VEGF-mRNA、VEGF표체급NaFl함량균현저고우NC조[VEGF-mRNA종(21.6±3.5)Kcopies/μg상승도(36.3 ±3.9)Kcopies/μg,VEGF종0.48±0.09증가도0.77±0.12,NaFl함량종(548±48)rfu/mg증가도(674±32)rfu/mg,균P＜0.01].여HC조상비,IVI조뇌조직VEGF mRNA、VEGF표체급NaFl함량차이무통계학의의(균P＞0.05).CVI조남조직VEGF-mRNA、VEGF표체급NaFl현저저우HC조[VEGF-mRNA종(36.3±3.9)Kcopies/μg강저도(19.9 ±4.3)Kcopies/μg,P＜0.01;VEGF종0.77±0.12강저도0.44 ±0.13,P＜0.01;NaFl함량종(674±32)rfu/mg하강도(542±77)rfu/mg,P＜0.05].각조간뇌조직함수솔차이무통계학의의(P＞0.05).결론 고원저양유발적VEGF표체증고시고원뇌수종적중요병리배절,뇌실내응용특이성침묵VEGF적siRNA가조단VEGF적표체,억제모세혈관통투성증고,가성위예방고원뇌수종적신도경.
Objective To observe the change of expression of vascular endothelial growth factor (VEGF)and vascular leakage in the brain of rats exposed to high altitude with siRNA targeting Vascular endothelial growth factor and explore the pathological mechanism and preventive approach of high altitude cerebral edema(HACE).Methods Fifty male Wistar rats were divided randomly into normal control group (Ncon),hish altitude control group(Hcon),intraventricular normal saline control group(Scon),intraventricular siRNA group(CVI)and intravenous siRNA group(IVI).Rats in Ncon were raised normally.Rats in Hcon,Scon,CVI and IVI pretreated with intravenous injection of normal saline,intraventricular injection of normal saline,intraventricular injection of siRNA and intravenous injection of siRNA respectively were exposed to a low-pressure cabin mimicking a high altitude of 7000 m for 24 h.The ratio of dry and wet brain weight was calculated and the sodium fluorescein leakage calculated to evaluate the cerebral edema and the blood brain barrier permeability.Also the real-time quantitative RT-PCR was employed to detect the expression of VEGF mRNA and the Western blot the expression of VEGF.Results Compared with rats in NC,high altitude exposure led to a significant increase in the levels of VEGF mRNA (from 21.6±3.5 Kcopies/μg to 36.3±3.9 Kcopies/μg,P＜0.01)and protein(from 48±0.09 to 0.77±0.12,P＜0.01)in rat brain and fluorescence intensity of sodium fluorescein increased significantly(from548±48 rfu/mg to 674±32 rfu/mg.P＜0.01).Intravenous injection of siRNA targeting to VEGF caused no significant change of expression VEGF mRNA and protein and fluorescence intensity of sodium fluorescein in rat brain(P＞0.05,respectively).While compared with rats in HC,intraventricular injection of siRNA targeting to VEGF caused the significant reduction of expression of VEGF mRNA (from 36.3±3.9 to 19.9±4.3,P＜0.01)and protein(from 0.77±0.12 to 0.44±0.13,P＜0.01)and fluorescence intensity of sodium fluorescein(from 674±32 rfu/mg to 542±77 rfu/mg,P＜0.05)in rat brain.There were no significant change in the ratio of dry and wet brain weight among five groups.Conclusion VEGF may play a key role in the pathologic process of HACE.Intraventricular injection of siRNA targeting to VEGF inhibits the expression of VEGF and prevent the high altitude-induced vascular leakage.These findings might provide a basis for new preventive approaches of cerebral edema.
