安徽医科大学学报
安徽醫科大學學報
안휘의과대학학보
Acta Universitatis Medicinalis Anhui
2015年
9期
1233-1237
,共5页
童晓琴%周方杰%蔡圣年%陈硕%陈志武%郭岩
童曉琴%週方傑%蔡聖年%陳碩%陳誌武%郭巖
동효금%주방걸%채골년%진석%진지무%곽암
内皮衍生超级化因子%硫化氢%局灶性脑缺血%大脑中动脉%血管舒张%超级化反应
內皮衍生超級化因子%硫化氫%跼竈性腦缺血%大腦中動脈%血管舒張%超級化反應
내피연생초급화인자%류화경%국조성뇌결혈%대뇌중동맥%혈관서장%초급화반응
endothelium-derived hyperpolarizing factor%H2 S%focal cerebral ischemia%MCA%vasodilation%hy-perpolarization
目的 探讨内皮衍生超极化因子( EDHF)对局灶性脑缺血再灌注(I/R)损伤大鼠大脑中动脉(MCA)的血管舒张作用以及对血管平滑肌细胞( VSMC)超级化反应的影响.方法 线栓法制备大鼠局灶性脑缺血( MCAO)再灌注损伤模型,按照Longa 法对实验动物进行神经功能缺陷评分;采用全自动酶标仪检测大鼠脑组织硫化氢( H2 S)含量的变化;取局灶性I/R损伤大鼠 MCA,显微镜下测量 MCA 的血管直径;用微电极记录MCA的VSMC的膜电位;RT-PCR 法测定I/R大鼠 MCA 内皮细胞胱硫醚-γ-裂解酶( CSE ) mRNA 表达.结果 I/R大鼠脑组织中的H2 S含量较正常大鼠明显增加;在离体I/R大鼠MCA,用3 × 10 -5 mol/L的 L-NAME (NO合酶抑制剂)和1 × 10 -5 mol/L的Indo(PGI2 合成酶抑制剂) 预灌30 min后,乙酰胆碱(ACh)(10 -7 ~10 -4.5 mol/L)可显著舒张血管,即 EDHF 介导的舒张反应明显增强;I/R大鼠 MCA 平滑肌细胞膜电位检测实验中,L-NAME 和 In-do 存在的条件下,ACh(10 -7 L~10 -4.5 mol/L )随着浓度增大,产生浓度依赖性的超极化作用,并且与正常大鼠MCA相比,超极化作用有明显的增强;I/R 大鼠 MCA 内皮细胞中CSE mRNA表达上调.结论 EDHF明显增强I/R大鼠MCA的血管舒张反应和平滑肌细胞超极化反应,表明内源性ED-HF( H2 S)对局灶性I/R损伤有保护作用.
目的 探討內皮衍生超極化因子( EDHF)對跼竈性腦缺血再灌註(I/R)損傷大鼠大腦中動脈(MCA)的血管舒張作用以及對血管平滑肌細胞( VSMC)超級化反應的影響.方法 線栓法製備大鼠跼竈性腦缺血( MCAO)再灌註損傷模型,按照Longa 法對實驗動物進行神經功能缺陷評分;採用全自動酶標儀檢測大鼠腦組織硫化氫( H2 S)含量的變化;取跼竈性I/R損傷大鼠 MCA,顯微鏡下測量 MCA 的血管直徑;用微電極記錄MCA的VSMC的膜電位;RT-PCR 法測定I/R大鼠 MCA 內皮細胞胱硫醚-γ-裂解酶( CSE ) mRNA 錶達.結果 I/R大鼠腦組織中的H2 S含量較正常大鼠明顯增加;在離體I/R大鼠MCA,用3 × 10 -5 mol/L的 L-NAME (NO閤酶抑製劑)和1 × 10 -5 mol/L的Indo(PGI2 閤成酶抑製劑) 預灌30 min後,乙酰膽堿(ACh)(10 -7 ~10 -4.5 mol/L)可顯著舒張血管,即 EDHF 介導的舒張反應明顯增彊;I/R大鼠 MCA 平滑肌細胞膜電位檢測實驗中,L-NAME 和 In-do 存在的條件下,ACh(10 -7 L~10 -4.5 mol/L )隨著濃度增大,產生濃度依賴性的超極化作用,併且與正常大鼠MCA相比,超極化作用有明顯的增彊;I/R 大鼠 MCA 內皮細胞中CSE mRNA錶達上調.結論 EDHF明顯增彊I/R大鼠MCA的血管舒張反應和平滑肌細胞超極化反應,錶明內源性ED-HF( H2 S)對跼竈性I/R損傷有保護作用.
목적 탐토내피연생초겁화인자( EDHF)대국조성뇌결혈재관주(I/R)손상대서대뇌중동맥(MCA)적혈관서장작용이급대혈관평활기세포( VSMC)초급화반응적영향.방법 선전법제비대서국조성뇌결혈( MCAO)재관주손상모형,안조Longa 법대실험동물진행신경공능결함평분;채용전자동매표의검측대서뇌조직류화경( H2 S)함량적변화;취국조성I/R손상대서 MCA,현미경하측량 MCA 적혈관직경;용미전겁기록MCA적VSMC적막전위;RT-PCR 법측정I/R대서 MCA 내피세포광류미-γ-렬해매( CSE ) mRNA 표체.결과 I/R대서뇌조직중적H2 S함량교정상대서명현증가;재리체I/R대서MCA,용3 × 10 -5 mol/L적 L-NAME (NO합매억제제)화1 × 10 -5 mol/L적Indo(PGI2 합성매억제제) 예관30 min후,을선담감(ACh)(10 -7 ~10 -4.5 mol/L)가현저서장혈관,즉 EDHF 개도적서장반응명현증강;I/R대서 MCA 평활기세포막전위검측실험중,L-NAME 화 In-do 존재적조건하,ACh(10 -7 L~10 -4.5 mol/L )수착농도증대,산생농도의뢰성적초겁화작용,병차여정상대서MCA상비,초겁화작용유명현적증강;I/R 대서 MCA 내피세포중CSE mRNA표체상조.결론 EDHF명현증강I/R대서MCA적혈관서장반응화평활기세포초겁화반응,표명내원성ED-HF( H2 S)대국조성I/R손상유보호작용.
Objective To explore the dilation and hyperpolarization induced by endothelium-derived hyperpolar-izing factor ( EDHF) in middle cerebral artery of rats subjected to cerebral ischemia/reperfusion ( I/R) . Methods The model of focal cerebral I/R injury in rat was established by middle cerebral artery occlusion ( MCAO) . Hy-drogen sulphide( H2S) level in the cerebrum was detected, and the dilation and resting membrane potential of VSMC induced by EDHF in MCA subjected to cerebral I/R were observed. RT-PCR method was used to detect the expression of cystathionine-γ-lyase ( CSE) mRNA in the endothelial cells of rat MCA. Results Compared with the control group, the contents of H2 S increased in the cerebrum of rat subjected to cerebral I/R. After 30 minutes per-fusion with PSS containing L-NAME (3 × 10 -5 mol/L) and Indo (1 × 10 -5 mol/L), the relaxant effects of ACh ( 10 -7 ~10 -4. 5 mol/L) on isolated rat MCAs subjected to cerebral I/R were more obvious by vasomotoricity experi-ment in vitro, namely, the vasodilatation induced by EDHF in cerebral I/R MCAs increased. In the presence of L-NAME (3 × 10 -5 mol/L) and Indo (1 × 10 -5 mol/L) , ACh(10 -7 ~10 -4. 5 mol/L) had stronger concentration de-pendent hyperpolarizing effect on the resting membrane potential of VSMC of cerebral I/R MCAs than the normal group. During cerebral I/R, the expression of CSEmRNA in rat MCAs endothelial cells was upregulated. Conclu-sion EDHF enhances dilation and hyperlarzation of VSMC in rat MCA subjected to cerebral I/R injury.
