临床神经病学杂志
臨床神經病學雜誌
림상신경병학잡지
Journal of Clinical Neurology
2015年
5期
356-361
,共6页
张瑞雪%吕欣欣%黄伟伟%池恒%唐吉友
張瑞雪%呂訢訢%黃偉偉%池恆%唐吉友
장서설%려흔흔%황위위%지항%당길우
聚腺苷二磷酸核糖聚合酶-1%RNA干扰%脑梗死%神经血管单元%超微结构
聚腺苷二燐痠覈糖聚閤酶-1%RNA榦擾%腦梗死%神經血管單元%超微結構
취선감이린산핵당취합매-1%RNA간우%뇌경사%신경혈관단원%초미결구
poly ( ADP-ribose ) polymerase-1%RNA interference%cerebral infarction%neurovascular unit%ultrastructure
目的 观察慢病毒介导的聚腺苷二磷酸核糖聚合酶-1(PARP-1)siRNA对大鼠脑梗死后神经血管单元(NVU)的影响. 方法 SD大鼠132只随机分为假手术组(n=32)、脑梗死组(n=30)、空病毒组(n=32)和PARP-1组(n=38). 空病毒组和PARP-1组大鼠分别于侧脑室注射空病毒和携带目的基因的病毒5μl进行RNA干扰,14 d后进行脑梗死模型制作. 造模术24 h后依据Longa 5分法进行神经功能评分. 采用TTC染色检测脑梗死体积,HE染色观察脑组织病理改变,伊文思蓝( EB)通透率检测计算脑组织EB含量,脑组织干湿重法测定脑组织含水量,电镜观察NVU超微结构的改变. 结果 假手术组与空病毒组间大鼠PARP-1 mRNA表达水平的差异无统计学意义(P=0.244). 与空病毒组比较, PARP-1组RNA干扰后3 d、5 d、8 d PARP-1 mRNA表达水平均明显下降(均P<0.05 ). 假手术组大鼠无神经功能缺损,无脑梗死. 与假手术组比较,脑梗死组和空病毒组大鼠缺血侧脑组织EB含量和脑组织含水量明显增加(均P<0.05 ). 与脑梗死组和空病毒组相比,PARP-1组大鼠神经功能评分显著降低,脑梗死体积明显减小,脑组织EB含量和脑组织含水量明显减少(均P<0.05). HE染色显示,假手术组大鼠海马神经细胞染色均匀,细胞排列紧密整齐且结构清晰;脑梗死组和空病毒组大鼠海马神经细胞肿胀、破裂、轮廓模糊,染色较深;PARP-1组神经细胞排列整齐,染色均匀. 电镜可见,假手术组海马NVU超微结构清晰完整;脑梗死组和空病毒组神经细胞变性,线粒体肿胀、嵴减少,髓鞘变薄、分层,血管内皮细胞凋亡、基底膜不连续,突触数量减少、结构破坏;而PARP-1组海马NVU超微结构损伤明显减轻. 结论 抑制PARP-1的表达,可明显改善脑梗死大鼠的神经功能,缩小脑梗死体积,减轻脑梗死后NVU的损伤.
目的 觀察慢病毒介導的聚腺苷二燐痠覈糖聚閤酶-1(PARP-1)siRNA對大鼠腦梗死後神經血管單元(NVU)的影響. 方法 SD大鼠132隻隨機分為假手術組(n=32)、腦梗死組(n=30)、空病毒組(n=32)和PARP-1組(n=38). 空病毒組和PARP-1組大鼠分彆于側腦室註射空病毒和攜帶目的基因的病毒5μl進行RNA榦擾,14 d後進行腦梗死模型製作. 造模術24 h後依據Longa 5分法進行神經功能評分. 採用TTC染色檢測腦梗死體積,HE染色觀察腦組織病理改變,伊文思藍( EB)通透率檢測計算腦組織EB含量,腦組織榦濕重法測定腦組織含水量,電鏡觀察NVU超微結構的改變. 結果 假手術組與空病毒組間大鼠PARP-1 mRNA錶達水平的差異無統計學意義(P=0.244). 與空病毒組比較, PARP-1組RNA榦擾後3 d、5 d、8 d PARP-1 mRNA錶達水平均明顯下降(均P<0.05 ). 假手術組大鼠無神經功能缺損,無腦梗死. 與假手術組比較,腦梗死組和空病毒組大鼠缺血側腦組織EB含量和腦組織含水量明顯增加(均P<0.05 ). 與腦梗死組和空病毒組相比,PARP-1組大鼠神經功能評分顯著降低,腦梗死體積明顯減小,腦組織EB含量和腦組織含水量明顯減少(均P<0.05). HE染色顯示,假手術組大鼠海馬神經細胞染色均勻,細胞排列緊密整齊且結構清晰;腦梗死組和空病毒組大鼠海馬神經細胞腫脹、破裂、輪廓模糊,染色較深;PARP-1組神經細胞排列整齊,染色均勻. 電鏡可見,假手術組海馬NVU超微結構清晰完整;腦梗死組和空病毒組神經細胞變性,線粒體腫脹、嵴減少,髓鞘變薄、分層,血管內皮細胞凋亡、基底膜不連續,突觸數量減少、結構破壞;而PARP-1組海馬NVU超微結構損傷明顯減輕. 結論 抑製PARP-1的錶達,可明顯改善腦梗死大鼠的神經功能,縮小腦梗死體積,減輕腦梗死後NVU的損傷.
목적 관찰만병독개도적취선감이린산핵당취합매-1(PARP-1)siRNA대대서뇌경사후신경혈관단원(NVU)적영향. 방법 SD대서132지수궤분위가수술조(n=32)、뇌경사조(n=30)、공병독조(n=32)화PARP-1조(n=38). 공병독조화PARP-1조대서분별우측뇌실주사공병독화휴대목적기인적병독5μl진행RNA간우,14 d후진행뇌경사모형제작. 조모술24 h후의거Longa 5분법진행신경공능평분. 채용TTC염색검측뇌경사체적,HE염색관찰뇌조직병리개변,이문사람( EB)통투솔검측계산뇌조직EB함량,뇌조직간습중법측정뇌조직함수량,전경관찰NVU초미결구적개변. 결과 가수술조여공병독조간대서PARP-1 mRNA표체수평적차이무통계학의의(P=0.244). 여공병독조비교, PARP-1조RNA간우후3 d、5 d、8 d PARP-1 mRNA표체수평균명현하강(균P<0.05 ). 가수술조대서무신경공능결손,무뇌경사. 여가수술조비교,뇌경사조화공병독조대서결혈측뇌조직EB함량화뇌조직함수량명현증가(균P<0.05 ). 여뇌경사조화공병독조상비,PARP-1조대서신경공능평분현저강저,뇌경사체적명현감소,뇌조직EB함량화뇌조직함수량명현감소(균P<0.05). HE염색현시,가수술조대서해마신경세포염색균균,세포배렬긴밀정제차결구청석;뇌경사조화공병독조대서해마신경세포종창、파렬、륜곽모호,염색교심;PARP-1조신경세포배렬정제,염색균균. 전경가견,가수술조해마NVU초미결구청석완정;뇌경사조화공병독조신경세포변성,선립체종창、척감소,수초변박、분층,혈관내피세포조망、기저막불련속,돌촉수량감소、결구파배;이PARP-1조해마NVU초미결구손상명현감경. 결론 억제PARP-1적표체,가명현개선뇌경사대서적신경공능,축소뇌경사체적,감경뇌경사후NVU적손상.
Objective To investigate the effects of Lentivirus-mediated poly ( ADP-ribose ) polymerase-1 ( PARP-1) siRNA transfer on neurovascular unit ( NVU) in rats following cerebral infarction ( CI) .Methods One hundred and thirty-two rats were randomly divided into sham group ( n=32 ) , CI group ( n=30 ) , control siRNA group (n=32), PARP-1 group (n=38).Lateral ventricle of rats in control siRNA group and PARP-1 group were injected with 5μl control siRNA or PARP-1 siRNA for RNA inference.CI model was performed 14 days later.Then, 24 h later, neurological evaluations were performed according to Longa five-point scale.TTC straining was conducted to assess CI volume.HE straining was used to examine cerebral pathological changes.Evans blue ( EB) leakage was conducted to assess the level of EB in brain.Brain water content was measured by dry-wet method.Electron microscopy was used to observe ultrastructure of NVU.Results No significant differences of PARP-1 mRNA were observed between sham group and control siRNA group (P=0.244).Compared with control siRNA group, PARP-1 mRNA was significantly decreased in PARP-1 group at 3 d, 5 d, 8 d after RNA interference ( all P<0.05 ) .No neurological deficits and CI were observed in sham group.Compared with sham group, EB leakage and brain water content were significantly increased in CI group and control siRNA group ( all P<0.05 ) .Compared with CI group and control siRNA group, neurobehavioral scores, CI volume, EB leakage and brain water content of PARP-1 group were significantly decreased ( all P<0 .0 5 ) .HE staining showed that neurons in hippocampus of sham group was stained evenly, cells were arranged orderly and apparented clearly;hippocampal neurons of the CI group and control siRNA group were swollen, fractured, indistinct and stained deeply;neurons in PARP-1 group were well-distributed and stained evenly.The electron microscope showed that ultrastructures of NVU in sham group were distinct and normal;neurons in the CI group and control siRNA group were denatured, mitochondria was swollen, mitochondrial cristae was fractured, myelin became thinner and delaminated, endothelialcell and basal lamina were deformed, synapse was reduced and destroyed; while ultrastructure deficits of NVU were ameliorated in PARP-1 group. Conclusion PARP-1 inhibition may improve neurological function, ameliorate CI volume, alleviate NVU deficits following CI.
