中华物理医学与康复杂志
中華物理醫學與康複雜誌
중화물리의학여강복잡지
CHINESE JOURNAL OF PHYSICAL MEDICINE AND REHABILITATION
2015年
7期
498-502
,共5页
电针预处理%脑缺血再灌注%血脑屏障%水通道蛋白-4%紧密连接蛋白-5
電針預處理%腦缺血再灌註%血腦屏障%水通道蛋白-4%緊密連接蛋白-5
전침예처리%뇌결혈재관주%혈뇌병장%수통도단백-4%긴밀련접단백-5
Acupuncture%Cerebral ischemia%Reperfusion%Blood-brain barrier%AQP4%CLN5
目的 观察不同时程电针(取穴百会、水沟)预处理对继发脑缺血再灌注大鼠血脑屏障水通道蛋白-4(AQP4)及紧密连接蛋白-5(CLN5)表达的影响.方法 采用随机数字表法将64只雄性SD大鼠分为模型组、假手术组、电针预处理7d组及电针预处理15d组,电针预处理7d组及电针预处理15d组分别给予持续7d、15d的电针(取穴百会、水沟)预处理.待上述预处理结束后,参照Longa改良线栓法将模型组、电针预处理7d组及电针预处理15d组大鼠制成单侧大脑中动脉栓塞(MCAO)模型,假手术组大鼠术中仅分离颈总动脉,不栓塞中动脉血流.各组大鼠于大脑中动脉栓塞90 min后进行再灌注.于制模24 h后采用免疫组化法、荧光定量PCR法检测各组大鼠血脑屏障AQP4、CLN5阳性细胞及其mRNA表达情况.结果 与假手术组比较,模型组及各电针预处理组大鼠血脑屏障AQP4阳性细胞及其mRNA表达均明显上调(P＜0.05),CLN5阳性细胞及其mRNA表达均明显下调(P＜0.05);与模型组比较,电针预处理7d组及15d组AQP4阳性细胞及其mRNA表达均明显下调(P＜0.05),CLN5阳性细胞及其mRNA表达均明显上调(P＜0.05);与电针预处理7d组比较,电针预处理15d组AQP4阳性细胞数[(25.15 ±0.55)个/每高倍镜视野]及其mRNA表达(1.16±0.04)下调幅度、CLN5阳性细胞数[(62.88±0.61)个/每高倍镜视野]及其mRNA表达(0.80±0.03)上调幅度均更显著(P＜0.05).结论 不同时程电针(取穴百会、水沟)预处理可抑制脑缺血再灌注大鼠血脑屏障AQP4阳性细胞及其mRNA表达,促进CLN5阳性细胞及其mRNA表达;以电针预处理15 d对脑缺血再灌注大鼠血脑屏障的保护作用相对较好;电针预处理诱导大鼠脑缺血耐受可能与调节脑缺血再灌后血脑屏障AQP4、CLN5及其mRNA表达有关.
目的 觀察不同時程電針(取穴百會、水溝)預處理對繼髮腦缺血再灌註大鼠血腦屏障水通道蛋白-4(AQP4)及緊密連接蛋白-5(CLN5)錶達的影響.方法 採用隨機數字錶法將64隻雄性SD大鼠分為模型組、假手術組、電針預處理7d組及電針預處理15d組,電針預處理7d組及電針預處理15d組分彆給予持續7d、15d的電針(取穴百會、水溝)預處理.待上述預處理結束後,參照Longa改良線栓法將模型組、電針預處理7d組及電針預處理15d組大鼠製成單側大腦中動脈栓塞(MCAO)模型,假手術組大鼠術中僅分離頸總動脈,不栓塞中動脈血流.各組大鼠于大腦中動脈栓塞90 min後進行再灌註.于製模24 h後採用免疫組化法、熒光定量PCR法檢測各組大鼠血腦屏障AQP4、CLN5暘性細胞及其mRNA錶達情況.結果 與假手術組比較,模型組及各電針預處理組大鼠血腦屏障AQP4暘性細胞及其mRNA錶達均明顯上調(P＜0.05),CLN5暘性細胞及其mRNA錶達均明顯下調(P＜0.05);與模型組比較,電針預處理7d組及15d組AQP4暘性細胞及其mRNA錶達均明顯下調(P＜0.05),CLN5暘性細胞及其mRNA錶達均明顯上調(P＜0.05);與電針預處理7d組比較,電針預處理15d組AQP4暘性細胞數[(25.15 ±0.55)箇/每高倍鏡視野]及其mRNA錶達(1.16±0.04)下調幅度、CLN5暘性細胞數[(62.88±0.61)箇/每高倍鏡視野]及其mRNA錶達(0.80±0.03)上調幅度均更顯著(P＜0.05).結論 不同時程電針(取穴百會、水溝)預處理可抑製腦缺血再灌註大鼠血腦屏障AQP4暘性細胞及其mRNA錶達,促進CLN5暘性細胞及其mRNA錶達;以電針預處理15 d對腦缺血再灌註大鼠血腦屏障的保護作用相對較好;電針預處理誘導大鼠腦缺血耐受可能與調節腦缺血再灌後血腦屏障AQP4、CLN5及其mRNA錶達有關.
목적 관찰불동시정전침(취혈백회、수구)예처리대계발뇌결혈재관주대서혈뇌병장수통도단백-4(AQP4)급긴밀련접단백-5(CLN5)표체적영향.방법 채용수궤수자표법장64지웅성SD대서분위모형조、가수술조、전침예처리7d조급전침예처리15d조,전침예처리7d조급전침예처리15d조분별급여지속7d、15d적전침(취혈백회、수구)예처리.대상술예처리결속후,삼조Longa개량선전법장모형조、전침예처리7d조급전침예처리15d조대서제성단측대뇌중동맥전새(MCAO)모형,가수술조대서술중부분리경총동맥,불전새중동맥혈류.각조대서우대뇌중동맥전새90 min후진행재관주.우제모24 h후채용면역조화법、형광정량PCR법검측각조대서혈뇌병장AQP4、CLN5양성세포급기mRNA표체정황.결과 여가수술조비교,모형조급각전침예처리조대서혈뇌병장AQP4양성세포급기mRNA표체균명현상조(P＜0.05),CLN5양성세포급기mRNA표체균명현하조(P＜0.05);여모형조비교,전침예처리7d조급15d조AQP4양성세포급기mRNA표체균명현하조(P＜0.05),CLN5양성세포급기mRNA표체균명현상조(P＜0.05);여전침예처리7d조비교,전침예처리15d조AQP4양성세포수[(25.15 ±0.55)개/매고배경시야]급기mRNA표체(1.16±0.04)하조폭도、CLN5양성세포수[(62.88±0.61)개/매고배경시야]급기mRNA표체(0.80±0.03)상조폭도균경현저(P＜0.05).결론 불동시정전침(취혈백회、수구)예처리가억제뇌결혈재관주대서혈뇌병장AQP4양성세포급기mRNA표체,촉진CLN5양성세포급기mRNA표체;이전침예처리15 d대뇌결혈재관주대서혈뇌병장적보호작용상대교호;전침예처리유도대서뇌결혈내수가능여조절뇌결혈재관후혈뇌병장AQP4、CLN5급기mRNA표체유관.
Objective To observe the effect of electroacupuncture (EA) pretreatment of different durations on the expressions of AQP4 and CLN5 in the blood-brain barrier after cerebral ischemia and reperfusion Methods Sixty-four male Sprague-Dawley (SD) rats were randomly divided into a model group,a sham operation group,a 7-day EA-pretreatment group (EAP-7 group) and a 15-day EA-pretreatment group (EAP-15 group),each of 16.After EA-pretreatment on the baihui and shuigou acupoints for 7 days or 15 days,a model of unilateral middle cerebral artery embolism (MCAO) model was induced in the rats of the model,EAP-7 and EAP-15 group using a modified Longa method.In the sham operation group the carotid artery was separated without middle cerebral artery embolism.The reperfusion was begun 90 min after the MCAO modeling.Immunohistochemical methods and fluorescence quantitative PCR were applied to detect the expression of AQP4,CLN5 and their mRNAs expression in the blood brain barrier (BBB) 24h after the operation.Results Compared with the sham operation group,the expression of AQP4 positive cells and AQP4 mRNA in the BBB in the other three groups had increased significantly,while the expression of CLN5 positive cells and CLN5 mRNA was significantly less.Compared with the model group,the expression of AQP4 positive cells and AQP4 mRNA in the EAP-7 and EAP-15 groups was significantly reduced,while the expression of CLN5 positive cells and CLN5 mRNA was significantly increased.Moreover,the expression of AQP4 positive cells and AQP4 mRNA in the EAP-15 group were significantly higher than in the EAP-7 group,while the expression of CLN5 positive cells and CLN5 mRNA were significantly lower.Conclusions EA-pretreatment on the baihui and shuigou acupoints can restrain the expression of AQP4-positive cells and AQP4 mRNA and promote that of CLN5-positive cells and CLN5 mRNA in the BBB after cerebral ischemia and reperfusion,at least in rats.The BBB protection effect is better when the EA-pretreatment lasts longer.The mechanisms of cerebral ischemia tolerance may be related to the regulation of AQP4,CLN5 and their mRNAs in the blood-brain barrier after cerebral injury.
