CAJ | 학술논문

目的研究乙酰胆碱( ACh)诱导内源性内皮衍生超极化因子( EDHF)对缺氧/再给氧大鼠海马神经细胞凋亡的影响。方法取原代培养的大鼠海马神经细胞制备缺氧/再给氧损伤模型；大鼠大脑中动脉( MCA)血管段用一氧化氮合酶抑制剂NG-nitro-L-argininemethyl ester ( L-NAME)和前列环素抑制剂indomethacin ( Indo)预处理，用ACh诱导血管内皮细胞合成释放内源性EDHF；分别用流式细胞仪法和Ho-echst荧光法检测海马神经元凋亡。结果流式细胞仪法和Hoechst荧光法检测均表明假缺氧组海马神经细胞的凋亡率很低，分别为3.1%和2.2%左右，而缺氧/再给氧的模型组细胞凋亡率有显著升高；与缺氧/再给氧模型组相比，内皮完整血管段合用1μmol/L ACh 或合用1μmol/L ACh、30μmol/L L-NAME及10μmol/L Indo均可明显抑制缺氧/再给氧所致的细胞凋亡，而单用ACh或内皮完整的血管或ACh合用去内皮的血管段对细胞凋亡无明显影响。结论 ACh诱导大鼠脑血管内皮细胞释放的内源性EDHF对缺氧/再给氧海马神经细胞凋亡有明显的抑制作用。
목적연구을선담감( ACh)유도내원성내피연생초겁화인자( EDHF)대결양/재급양대서해마신경세포조망적영향。방법취원대배양적대서해마신경세포제비결양/재급양손상모형；대서대뇌중동맥( MCA)혈관단용일양화담합매억제제NG-nitro-L-argininemethyl ester ( L-NAME)화전렬배소억제제indomethacin ( Indo)예처리，용ACh유도혈관내피세포합성석방내원성EDHF；분별용류식세포의법화Ho-echst형광법검측해마신경원조망。결과류식세포의법화Hoechst형광법검측균표명가결양조해마신경세포적조망솔흔저，분별위3.1%화2.2%좌우，이결양/재급양적모형조세포조망솔유현저승고；여결양/재급양모형조상비，내피완정혈관단합용1μmol/L ACh 혹합용1μmol/L ACh、30μmol/L L-NAME급10μmol/L Indo균가명현억제결양/재급양소치적세포조망，이단용ACh혹내피완정적혈관혹ACh합용거내피적혈관단대세포조망무명현영향。결론 ACh유도대서뇌혈관내피세포석방적내원성EDHF대결양/재급양해마신경세포조망유명현적억제작용。
Objective To study the effect of acetylcholine ( ACh) induced endogenous endothelium-derived hyper-polarizing factor ( EDHF ) on apoptosis of rat hippocampal neurons. Methods Primarily cultured hippocampal neu-rons were insulted by hypoxia/reoxygenation ( H/R) , and EDHF from rat MCA segment was induced by acetylcho-line ( ACh) in the presence of NG-nitro-L-argininemethyl ester ( L-NAME) and indomethacin ( Indo) , the rate of hippocampal neuron apoptosis was detected by Hoechst staining and flow cytometry ( FC) methods. Results Ex-aminations of FC and Hoechst staining showed that the apoptosic rate of rat hippocampal neurons was very low in the sham group, only about 3. 1% and 2. 2% respectively,but H/R injury caused an obvious increase of apoptosis of rat hippocampal neurons. Co-adminstration of endothelium-intact MCA segment with 1 μmol/L ACh or co-admin-stration of endothelium-intact MCA segment with 1 μmol/L ACh + 30 μmol/L L-NAME + 10 μmol/L Indo markedly inhibits H/R-induced apoptosis of rat hippocamal neurons. However, administration of ACh or MCA seg-ment separately had not effect on H/R-induced apoptosis of rat hippocamal neurons, and neither did co-adminstra-tion of endothelium-denuded MCA segment with ACh. Conclusion Endogenous EDHF released from ACh-induced vascular endothelial cells significantly inhibits H/R-caused apoptosis of rat primarily cultured neurons.