CAJ | 학술논문

目的探讨给予外源性硫化氢(H2S)供体硫氢化钠(NaHS)对氯化锂-匹罗卡品致癫痫持续状态幼鼠海马神经元损伤的影响. 方法 3～4周龄SD大鼠按随机数字表法分为5组:正常组、正常+NaHS组、癫痫组、癫痫+NaHS组、癫痫+羟胺组.后3组大鼠应用腹腔注射氯化锂+阿托品溶液+盐酸匹罗卡品溶液方法建立癫痫持续状态模型,并分别给予NaHS 10 mg/kg或羟胺1mg/kg或等量生理盐水腹腔注射.干预后7d各组取6只大鼠收集海马组织及血浆,采用分光光度法测定血浆及海马H2S浓度,采用RT-PCR检测海马胱硫醚β-合酶(CBS) mRNA表达水平,采用Nissl染色观察海马神经元形态,采用Western blotting检测海马组织神经元特异性烯醇化酶(NSE)蛋白表达水平. 结果分光光度法和RT-PCR检测结果显示:血浆和海马H2S浓度及CBSmRNA表达水平在癫痫组均明显低于正常组,在癫痫+NaHS组均显著高于癫痫组,而在癫痫+羟胺组均显著低于癫痫组,差异均有统计学意义(P＜0.05).Nissl染色结果显示:癫痫组海马CA3区尼氏体明显减少,锥体细胞排列紊乱,而癫痫+NaHS组海马尼氏体缺失及锥体细胞排列紊乱较癫痫组明显减轻.Western blotting结果显示:海马组织中NSE表达水平在癫痫组显著高于正常组,在癫痫+NaHS组显著低于癫痫组,而在癫痫+羟胺组显著高于癫痫组,差异均有统计学意义(P＜0.05).结论给予外源性H2S供体NaHS能够显著上调海马组织CBS mRNA表达水平,提高血浆及海马组织H2S含量,降低海马NSE表达水平,从而改善癫痫所致的海马神经元损伤.
목적 탐토급여외원성류화경(H2S)공체류경화납(NaHS)대록화리-필라잡품치전간지속상태유서해마신경원손상적영향. 방법 3～4주령SD대서안수궤수자표법분위5조:정상조、정상+NaHS조、전간조、전간+NaHS조、전간+간알조.후3조대서응용복강주사록화리+아탁품용액+염산필라잡품용액방법건립전간지속상태모형,병분별급여NaHS 10 mg/kg혹간알1mg/kg혹등량생리염수복강주사.간예후7d각조취6지대서수집해마조직급혈장,채용분광광도법측정혈장급해마H2S농도,채용RT-PCR검측해마광류미β-합매(CBS) mRNA표체수평,채용Nissl염색관찰해마신경원형태,채용Western blotting검측해마조직신경원특이성희순화매(NSE)단백표체수평. 결과 분광광도법화RT-PCR검측결과현시:혈장화해마H2S농도급CBSmRNA표체수평재전간조균명현저우정상조,재전간+NaHS조균현저고우전간조,이재전간+간알조균현저저우전간조,차이균유통계학의의(P＜0.05).Nissl염색결과현시:전간조해마CA3구니씨체명현감소,추체세포배렬문란,이전간+NaHS조해마니씨체결실급추체세포배렬문란교전간조명현감경.Western blotting결과현시:해마조직중NSE표체수평재전간조현저고우정상조,재전간+NaHS조현저저우전간조,이재전간+간알조현저고우전간조,차이균유통계학의의(P＜0.05).결론 급여외원성H2S공체NaHS능구현저상조해마조직CBS mRNA표체수평,제고혈장급해마조직H2S함량,강저해마NSE표체수평,종이개선전간소치적해마신경원손상.
Objective To explore the effects of exogenous supplier of hydrogen sulfide (H2S) sodium hydrosulfide (NaHS) on hippocampal neuronal damage in immature rats with status epilepticus (SE) induced by lithium-pilocarpine.Methods Three or four weeks old Sprague Dawley rats were divided into five groups according to random number table:normal group,normal+NaHS group,epilepsy group,epilepsy+NaHS group and epilepsy+hydroxylamine (HA) group (n=6).SE models were induced by lithium-pilocarpine in immature rats of epilepsy group,epilepsy+NaHS group and epilepsy+HA group;meanwhile,NaHS (10 mg/kg) or hydroxylamine (1 mg/kg) or equivalent normal saline were given by intraperitoneal injection,respectively.Hippocampus tissues and plasma were collected,and the concentrations of H2S in the plasma and hippocampus were detected by spectrophotometer.The cystathionine beta synthase (CBS) mRNA expression in the hippocampus was assayed by real-time PCR.Hippocampal neurons were observed by Nissl staining.The neuron-specific enolase (NSE) expression in the hippocampus was assayed by Western blotting.Results Spectrophotometer and real-time PCR results showed that the concentration of HzS in the plasma and hippocampus and the CBS mRNA levels in the hippocampus of the epilepsy group were significantly lower than those in the normal group (P＜0.05);these indexes in the epilepsy+NaHS group were up-regulated as compared with those in the epilepsy group,while those in the epilepsy+HA group were significantly lower than those in the epilepsy group (P＜0.05).Nissl staining showed that decreased number of nissl bodies and disordered arrangement of pyramidal cells in the hippocampus CA3 area of epilepsy group were noted as compared with those in the normal group;the loss of nissl bodies and the disordered arrangement of pyramidal ceils in the hippocampus in epilepsy+NaHS group were improved as compared with those in the epilepsy group.Western blotting showed that the NSE expression in the epilepsy group was significantly higher than that in normal group,and NSE expression in epilepsy+NaHS group was significantly lower than that in the epilepsy group,while NSE expression in epilepsy+HA group was significantly higher than that in the epilepsy group (P＜0.05).Conclusion Exogenous H2S supplier NaHS could significantly up-regulate the CBS mRNA level in the hippocampus,increase the hydrogen sulfide content in the plasma and hippocampus,decrease the NSE expression in the hippocampus,and eventually attenuate neurons damage of hippocampus caused by epilepsy in rats.