中华神经科杂志
中華神經科雜誌
중화신경과잡지
Chinese Journal of Neurology
2015年
1期
55-60
,共6页
梗死,大脑中动脉%神经再生%新生血管化,病理性%激肽释放酶类%微气泡
梗死,大腦中動脈%神經再生%新生血管化,病理性%激肽釋放酶類%微氣泡
경사,대뇌중동맥%신경재생%신생혈관화,병이성%격태석방매류%미기포
Infarction,middle cerebral artery%Nerve regeneration%Neovascularization,pathologic%Kallikreins%Microbubbles
目的 探讨超声介导微泡携带激肽原酶靶向治疗技术对大鼠急性脑梗死后神经再生及血管新生的影响.方法 线栓法建立局灶性大脑中动脉闭塞(MCAO)模型,按随机数字表法分为超声介导激肽原酶微泡组、超声介导空白微泡对照组、超声辐照对照组、激肽原酶对照组、生理盐水对照组,每组24只.机械振荡法制备激肽原酶载药微泡.MCAO术后24h所有大鼠按不同组别予以尾静脉给药,每天1次,连续给药2d或6d.超声组在尾静脉给药的同时,予以超声辐照缺血侧颅骨10 min.分别在第3天和第7天行神经功能评分,并处死大鼠,测量脑梗死体积,观察侧脑室室管膜下区(SVZ)双皮质素的表达变化和梗死灶周围血管密度层黏连蛋白表达的变化.结果 术后7d超声介导激肽原酶微泡组、超声介导空白微泡对照组、超声辐照对照组、激肽原酶对照组、生理盐水对照组SVZ区在每20倍目镜视野下双皮质素阳性细胞数分别为251.8 ±13.1、125.7±11.6、130.2±13.7、234.5±12.4和123.7±10.0,梗死灶周围区在每10倍目镜视野下层黏连蛋白免疫阳性百分比分别为10.0％±0.8％、5.2％±0.7％、5.0％±1.0％、8.0％±1.8％和5.0％±0.9％.其中,超声介导激肽原酶微泡组和激肽原酶组梗死侧SVZ区双皮质素阳性细胞数和梗死灶周围层黏连蛋白免疫阳性百分比均明显高于同时点超声介导空白微泡对照组、超声辐照对照组、生理盐水对照组(双皮质素:超声介导激肽原酶微泡组比超声介导空白微泡对照组、超声辐照对照组、生理盐水对照组,t=17.88、17.17、18.16,P＜0.01;激肽原酶组比超声介导空白微泡对照组、超声辐照对照组、生理盐水对照组,t=15.42、14.78、15.70,P＜0.01.层黏连蛋白:超声介导激肽原酶微泡组比超声介导空白微泡对照组、超声辐照对照组、生理盐水对照组,t=7.01、6.71、7.11,P＜0.01;激肽原酶组比超声介导空白微泡对照组、超声辐照对照组、生理盐水对照组,t=4.23、3.94、4.33,P＜0.01).另外,超声介导激肽原酶微泡组与激肽原酶对照组相比,梗死侧SVZ区双皮质素阳性细胞数(t=2.46,P＜0.05)及梗死灶周围层黏连蛋白免疫阳性百分比(t =2.78,P＜0.05)均更高,差异有统计学意义.MCAO术后7d上述各组的神经功能评分超声介导激肽原酶微泡组与其他4组相比,差异均有统计学意义.结论 超声介导激肽原酶载药微泡治疗较单用激肽原酶治疗更能促进大鼠MCAO术后的神经再生和血管新生,进一步改善大鼠急性脑梗死后的神经功能,因此,超声介导载药微泡靶向治疗技术有望进一步用于神经系统疾病的治疗.
目的 探討超聲介導微泡攜帶激肽原酶靶嚮治療技術對大鼠急性腦梗死後神經再生及血管新生的影響.方法 線栓法建立跼竈性大腦中動脈閉塞(MCAO)模型,按隨機數字錶法分為超聲介導激肽原酶微泡組、超聲介導空白微泡對照組、超聲輻照對照組、激肽原酶對照組、生理鹽水對照組,每組24隻.機械振盪法製備激肽原酶載藥微泡.MCAO術後24h所有大鼠按不同組彆予以尾靜脈給藥,每天1次,連續給藥2d或6d.超聲組在尾靜脈給藥的同時,予以超聲輻照缺血側顱骨10 min.分彆在第3天和第7天行神經功能評分,併處死大鼠,測量腦梗死體積,觀察側腦室室管膜下區(SVZ)雙皮質素的錶達變化和梗死竈週圍血管密度層黏連蛋白錶達的變化.結果 術後7d超聲介導激肽原酶微泡組、超聲介導空白微泡對照組、超聲輻照對照組、激肽原酶對照組、生理鹽水對照組SVZ區在每20倍目鏡視野下雙皮質素暘性細胞數分彆為251.8 ±13.1、125.7±11.6、130.2±13.7、234.5±12.4和123.7±10.0,梗死竈週圍區在每10倍目鏡視野下層黏連蛋白免疫暘性百分比分彆為10.0％±0.8％、5.2％±0.7％、5.0％±1.0％、8.0％±1.8％和5.0％±0.9％.其中,超聲介導激肽原酶微泡組和激肽原酶組梗死側SVZ區雙皮質素暘性細胞數和梗死竈週圍層黏連蛋白免疫暘性百分比均明顯高于同時點超聲介導空白微泡對照組、超聲輻照對照組、生理鹽水對照組(雙皮質素:超聲介導激肽原酶微泡組比超聲介導空白微泡對照組、超聲輻照對照組、生理鹽水對照組,t=17.88、17.17、18.16,P＜0.01;激肽原酶組比超聲介導空白微泡對照組、超聲輻照對照組、生理鹽水對照組,t=15.42、14.78、15.70,P＜0.01.層黏連蛋白:超聲介導激肽原酶微泡組比超聲介導空白微泡對照組、超聲輻照對照組、生理鹽水對照組,t=7.01、6.71、7.11,P＜0.01;激肽原酶組比超聲介導空白微泡對照組、超聲輻照對照組、生理鹽水對照組,t=4.23、3.94、4.33,P＜0.01).另外,超聲介導激肽原酶微泡組與激肽原酶對照組相比,梗死側SVZ區雙皮質素暘性細胞數(t=2.46,P＜0.05)及梗死竈週圍層黏連蛋白免疫暘性百分比(t =2.78,P＜0.05)均更高,差異有統計學意義.MCAO術後7d上述各組的神經功能評分超聲介導激肽原酶微泡組與其他4組相比,差異均有統計學意義.結論 超聲介導激肽原酶載藥微泡治療較單用激肽原酶治療更能促進大鼠MCAO術後的神經再生和血管新生,進一步改善大鼠急性腦梗死後的神經功能,因此,超聲介導載藥微泡靶嚮治療技術有望進一步用于神經繫統疾病的治療.
목적 탐토초성개도미포휴대격태원매파향치료기술대대서급성뇌경사후신경재생급혈관신생적영향.방법 선전법건립국조성대뇌중동맥폐새(MCAO)모형,안수궤수자표법분위초성개도격태원매미포조、초성개도공백미포대조조、초성복조대조조、격태원매대조조、생리염수대조조,매조24지.궤계진탕법제비격태원매재약미포.MCAO술후24h소유대서안불동조별여이미정맥급약,매천1차,련속급약2d혹6d.초성조재미정맥급약적동시,여이초성복조결혈측로골10 min.분별재제3천화제7천행신경공능평분,병처사대서,측량뇌경사체적,관찰측뇌실실관막하구(SVZ)쌍피질소적표체변화화경사조주위혈관밀도층점련단백표체적변화.결과 술후7d초성개도격태원매미포조、초성개도공백미포대조조、초성복조대조조、격태원매대조조、생리염수대조조SVZ구재매20배목경시야하쌍피질소양성세포수분별위251.8 ±13.1、125.7±11.6、130.2±13.7、234.5±12.4화123.7±10.0,경사조주위구재매10배목경시야하층점련단백면역양성백분비분별위10.0％±0.8％、5.2％±0.7％、5.0％±1.0％、8.0％±1.8％화5.0％±0.9％.기중,초성개도격태원매미포조화격태원매조경사측SVZ구쌍피질소양성세포수화경사조주위층점련단백면역양성백분비균명현고우동시점초성개도공백미포대조조、초성복조대조조、생리염수대조조(쌍피질소:초성개도격태원매미포조비초성개도공백미포대조조、초성복조대조조、생리염수대조조,t=17.88、17.17、18.16,P＜0.01;격태원매조비초성개도공백미포대조조、초성복조대조조、생리염수대조조,t=15.42、14.78、15.70,P＜0.01.층점련단백:초성개도격태원매미포조비초성개도공백미포대조조、초성복조대조조、생리염수대조조,t=7.01、6.71、7.11,P＜0.01;격태원매조비초성개도공백미포대조조、초성복조대조조、생리염수대조조,t=4.23、3.94、4.33,P＜0.01).령외,초성개도격태원매미포조여격태원매대조조상비,경사측SVZ구쌍피질소양성세포수(t=2.46,P＜0.05)급경사조주위층점련단백면역양성백분비(t =2.78,P＜0.05)균경고,차이유통계학의의.MCAO술후7d상술각조적신경공능평분초성개도격태원매미포조여기타4조상비,차이균유통계학의의.결론 초성개도격태원매재약미포치료교단용격태원매치료경능촉진대서MCAO술후적신경재생화혈관신생,진일보개선대서급성뇌경사후적신경공능,인차,초성개도재약미포파향치료기술유망진일보용우신경계통질병적치료.
Objective To evaluate the influence of ultrasound-mediated contrast agent microbubbles carrying kallidinogenase targeted therapy on neurogenesis and angiogenesis after experimental acute cerebral infarction.Methods Kallidinogenase-loaded microbubbles were prepared using mechanical shaking method.Middle cerebral artery occlusion (MCAO) model was established in male Wistar rats by sutureoccluded method.MCAO rats (n =120) were randomly divided into 5 groups:ultrasound-mediated kallidinogenase-loaded microbubbles group (group 1),ultrasound-mediated microbubbles group (group 2),ultrasound-mediated saline group (group 3),kallidinogenase group (group 4),saline group (group 5).Medication was given through tail vein daily for 2-6 consecutive days starting 24 h after MCAO.Ultrasound groups (groups 1,2 and 3) were given 2 MHz pulse ultrasonic irradiation on the ischemia lateral skull for 10 min after injection.Cell proliferation was examined using 5'-bromo-2'-deoxyuridine (BrdU,50 mg/kg).Infarction volume and neurological function were evaluated on the 3rd,7th day after MCAO respectively.Doublecortin (DCX) + cells in the subventricular zone and laminin + in the peri-infarction region were observed at the same time.Results The number of DCX + cells in the subventricular zone (under 20 × ocular) of groups 1-5 was 251.8 ± 13.1,125.7 ± 11.6,130.2 ± 13.7,234.5 ± 12.4 and 123.7 ± 10.0 respectively.The percentage of laminin + cells in the peri-infarction region (× 10 objective) of groups 1-5 was 10.0％ ± 0.8％,5.2％ ± 0.7％,5.0％ ± 1.0％,8.0％ ± 1.8％ and 5.0％ ± 0.9％ respectively.The number of DCX + cells in the subventricular zone and laminin + cells in the peri-infarction region of the group 1 and the group 4 was significantly more on the 7th day after MCAO compared with those of the other three groups (DCX:t values of group 1 vs groups 2,3,5 were 17.88,17.17 and 18.16,all P ＜ 0.01 ; t values of group 4 vs groups 2,3,5 were 15.42,14.78 and 15.70,all P ＜0.01.Laminin:t values of group 1 vs groups 2,3,5 were 7.01,6.71 and 7.11,all P ＜ 0.01 ; t values of group 4 vs groups 2,3,5 were 4.23,3.94 and 4.33,all P ＜0.01).Moreover,the number of DCX+ cells in the subventricular zone (t =2.46,P ＜ 0.05) and laminin + cells in the peri-infarction region (t =2.78,P ＜ 0.05) of the group 1 was much more than those of the group 4.Neurologic scores on the 7th day of groups 1-5 were 1.00 (0.75,1.25),2.0 (2.00,3.00),2.0 (1.00,2.00),1.5 (0.75,2.00),2.0 (2.00,2.50).Compared with other four groups,group 1 showed better functional improvement after stroke (U values of group 1 vs groups 2-5 were 2.0,4.0,7.5 and 2.5,all P ＜ 0.05).While there was no significant difference in infarction volume among five groups at all the time points after MCAO.Conclusions Our study demonstrates ultrasound-mediated kallidinogenase-loaded contrast agent microbubbles targeted therapy promotes neuroblasts proliferation and vascular regeneration compared with mediated and non-medicine-loaded microbubbles therapy,which attributes to functional improvement after MCAO.Therefore,ultrasound-mediated ultrasound contrast agent microbubbles carrying drugs targeted therapy may have a perspective on ischemic stroke.
