目的 评价乳果糖对大鼠脊髓缺血再灌注损伤的影响.方法 选择胃管置入成功的健康雄性清洁级SD大鼠144只,3月龄,体重300～ 350 g,采用随机数字表法,将其分为4组(n=36):假手术组(S组)、脊髓缺血再灌注组(I/R组)、乳果糖组(L组)和乳果糖+抗生素组(LA组).采用阻断胸主动脉联合体循环低血压的方法诱导脊髓缺血9 min,然后恢复灌注,建立脊髓缺血再灌注损伤模型.L组于再灌注即刻胃内灌注乳果糖0.5 g/kg.LA组于术前1～3d胃内灌注甲硝唑和庆大霉素,每日单次剂量分别为30和40 mg/kg,3次/d,余处理同L组.于缺血前、再灌注10、30、60、90、120、150和180 min时采用氢气探针测定脑脊液氢气浓度.于再灌注12、24和48 h时,采用Westernblot法测定脊髓核因子E2相关因子-2(Nrf2)表达水平,黄嘌呤氧化酶法测定超氧化物歧化酶(SOD)活性,钼酸铵法测定过氧化氢酶(CAT)活性,ELISA法测定脊髓8-羟基脱氧鸟苷酸(8-OHdG)、3-硝基酪氨酸(3-NT)和丙二醛(MDA)的含量.于再灌注48 h时行后肢神经行为学评分,然后处死大鼠,取L3-5脊髓组织,测定神经元存活率和凋亡率.结果 与S组比较,I/R组再灌注各时点脑脊液氢气浓度差异无统计学意义,再灌注12、24和48 h时脊髓Nrf2、CAT和SOD水平差异无统计学意义(P＞0.05),8-OHdG、3-NT、MDA含量增加,后肢神经行为学评分和神经元存活率降低,神经元凋亡率升高,L组再灌注30～ 180 min时脑脊液氢气浓度升高,再灌注12、24和48 h时脊髓Nrf2、CAT和SOD水平增加(P＜0.05),8-OHdG、3-NT、MDA含量、后肢神经行为学评分、神经元存活率和凋亡率差异无统计学意义(P＞0.05).与I/R组比较,L组再灌注30～ 180 min时脑脊液氢气浓度升高,脊髓Nrf2、CAT和SOD水平增加,8-OHdG、3-NT和MDA含量降低,后肢神经行为学评分和神经元存活率升高,神经元凋亡率降低(P＜0.05),LA组上述指标差异无统计学意义(P＞0.05).结论 乳果糖可减轻大鼠脊髓缺血再灌注损伤.
目的 評價乳果糖對大鼠脊髓缺血再灌註損傷的影響.方法 選擇胃管置入成功的健康雄性清潔級SD大鼠144隻,3月齡,體重300～ 350 g,採用隨機數字錶法,將其分為4組(n=36):假手術組(S組)、脊髓缺血再灌註組(I/R組)、乳果糖組(L組)和乳果糖+抗生素組(LA組).採用阻斷胸主動脈聯閤體循環低血壓的方法誘導脊髓缺血9 min,然後恢複灌註,建立脊髓缺血再灌註損傷模型.L組于再灌註即刻胃內灌註乳果糖0.5 g/kg.LA組于術前1～3d胃內灌註甲硝唑和慶大黴素,每日單次劑量分彆為30和40 mg/kg,3次/d,餘處理同L組.于缺血前、再灌註10、30、60、90、120、150和180 min時採用氫氣探針測定腦脊液氫氣濃度.于再灌註12、24和48 h時,採用Westernblot法測定脊髓覈因子E2相關因子-2(Nrf2)錶達水平,黃嘌呤氧化酶法測定超氧化物歧化酶(SOD)活性,鉬痠銨法測定過氧化氫酶(CAT)活性,ELISA法測定脊髓8-羥基脫氧鳥苷痠(8-OHdG)、3-硝基酪氨痠(3-NT)和丙二醛(MDA)的含量.于再灌註48 h時行後肢神經行為學評分,然後處死大鼠,取L3-5脊髓組織,測定神經元存活率和凋亡率.結果 與S組比較,I/R組再灌註各時點腦脊液氫氣濃度差異無統計學意義,再灌註12、24和48 h時脊髓Nrf2、CAT和SOD水平差異無統計學意義(P＞0.05),8-OHdG、3-NT、MDA含量增加,後肢神經行為學評分和神經元存活率降低,神經元凋亡率升高,L組再灌註30～ 180 min時腦脊液氫氣濃度升高,再灌註12、24和48 h時脊髓Nrf2、CAT和SOD水平增加(P＜0.05),8-OHdG、3-NT、MDA含量、後肢神經行為學評分、神經元存活率和凋亡率差異無統計學意義(P＞0.05).與I/R組比較,L組再灌註30～ 180 min時腦脊液氫氣濃度升高,脊髓Nrf2、CAT和SOD水平增加,8-OHdG、3-NT和MDA含量降低,後肢神經行為學評分和神經元存活率升高,神經元凋亡率降低(P＜0.05),LA組上述指標差異無統計學意義(P＞0.05).結論 乳果糖可減輕大鼠脊髓缺血再灌註損傷.
목적 평개유과당대대서척수결혈재관주손상적영향.방법 선택위관치입성공적건강웅성청길급SD대서144지,3월령,체중300～ 350 g,채용수궤수자표법,장기분위4조(n=36):가수술조(S조)、척수결혈재관주조(I/R조)、유과당조(L조)화유과당+항생소조(LA조).채용조단흉주동맥연합체순배저혈압적방법유도척수결혈9 min,연후회복관주,건립척수결혈재관주손상모형.L조우재관주즉각위내관주유과당0.5 g/kg.LA조우술전1～3d위내관주갑초서화경대매소,매일단차제량분별위30화40 mg/kg,3차/d,여처리동L조.우결혈전、재관주10、30、60、90、120、150화180 min시채용경기탐침측정뇌척액경기농도.우재관주12、24화48 h시,채용Westernblot법측정척수핵인자E2상관인자-2(Nrf2)표체수평,황표령양화매법측정초양화물기화매(SOD)활성,목산안법측정과양화경매(CAT)활성,ELISA법측정척수8-간기탈양조감산(8-OHdG)、3-초기락안산(3-NT)화병이철(MDA)적함량.우재관주48 h시행후지신경행위학평분,연후처사대서,취L3-5척수조직,측정신경원존활솔화조망솔.결과 여S조비교,I/R조재관주각시점뇌척액경기농도차이무통계학의의,재관주12、24화48 h시척수Nrf2、CAT화SOD수평차이무통계학의의(P＞0.05),8-OHdG、3-NT、MDA함량증가,후지신경행위학평분화신경원존활솔강저,신경원조망솔승고,L조재관주30～ 180 min시뇌척액경기농도승고,재관주12、24화48 h시척수Nrf2、CAT화SOD수평증가(P＜0.05),8-OHdG、3-NT、MDA함량、후지신경행위학평분、신경원존활솔화조망솔차이무통계학의의(P＞0.05).여I/R조비교,L조재관주30～ 180 min시뇌척액경기농도승고,척수Nrf2、CAT화SOD수평증가,8-OHdG、3-NT화MDA함량강저,후지신경행위학평분화신경원존활솔승고,신경원조망솔강저(P＜0.05),LA조상술지표차이무통계학의의(P＞0.05).결론 유과당가감경대서척수결혈재관주손상.
Objective To evaluate the effect of lactulose on spinal cord ischemia-reperfusion (I/R) injury in rats.Methods One hundred forty-four male Sprague-Dawley rats, aged 3 months, weighing 300-350 g, in which gastric tube was successfully inserted, were randomly divided into 4 groups (n=36 each) using a random number table: sham operation group (group S), spinal cord I/R group (group I/R), lactulose group (group L), and lactulose + antibiotics group (group LA).Spinal cord ischemia was induced by occlusion of the thoracic aorta combined with controlled hypotension for 9 min, followed by reperfusion.Lactulose 0.5 g/kg was administered intragastrically immediately after onset of reperfusion in group L.Metronidazole 30 mg/kg and gentamicin 40 mg/kg were administered intragastrically three times a day during 1-3 days before operation in group LA, and the other procedures were similar to those previously described in group L.Hydrogen concentration in cerebrospinal fluid was detected before ischemia and at 10, 30, 60, 90, 120, 150 and 180 min after reperfusion.At 12, 24 and 48 h of reperfusion, 8 rats in each group were sacrificed, and the L3-5 segment of the spinal cord was isolated for determination of nuclear factor E2-related factor 2 (Nrf2) expression (by Western blot), superoxide dismutase (SOD) activity (by using xanthine oxidase method), catalase (CAT) activity (ammonium molybdate method), and 8-hydroxy-2'-deoxyguanosine (8-OHdG), 3-nitrotyrosine (3-NT) and malondialdehyde (MDA) contents (by ELISA).Neurological function was assessed and scored at 48 h of reperfusion.Six animals in each group were then sacrificed after assessment of neurological function, and the L3-5 segment of the spinal cord was removed for detection of apoptotic neurons.The cell survival rate and apoptotic rate were calculated.Results Compared with group S, no significant change was found in the hydrogen concentration in the cerebrospinal fluid at each time point, and in the level of Nrf2, CAT and SOD at 12, 24 and 48 h of reperfusion, the contents of 8-OHdG, 3-NT and MDA were significantly increased, the neurological scores and cell survival rate were decreased, and the apoptotic rate was increased in group I/R, and the hydrogen concentration in the cerebrospinal fluid at 30-180 min of reperfusion was increased, the level of Nrf2, CAT and SOD was increased at 12, 24 and 48 h of reperfusion, and no significant change was found in the contents of 8-OHdG, 3-NT and MDA, neurological scores, cell survival rate, and apoptotic rate in group L.Compared with group I/R, the hydrogen concentration in the cerebrospinal fluid at 30-180 min of reperfusion was increased, the level of Nrf2, CAT and SOD was increased, the contents of 8-OHdG, 3-NT and MDA were decreased, the neurological scores and cell survival rate were increased, and the apoptotic rate was decreased in group L, and no significant change was found in the parameters mentioned above in group LA.Conclusion Lactulose can reduce spinal cord I/R injury in rats.
