背景:山莨菪碱是从茄科唐古特莨菪中提取的一种生物碱,是一种良好的细胞保护剂.而线粒体功能变化是反映细胞损伤的最敏感指标之一.目的:观察山莨菪碱对家兔全脑缺血再灌流后脑线粒体损伤的影响,探讨山莨菪碱的脑保护作用.设计:完全随机对照实验.单位:华中科技大学同济医学院附属同济医院.材料:实验于2002-09/12在华中科技大学同济医学院附属同济医院急诊科实验室完成.选择30只健康家兔,随机分为假手术组、缺血再灌流组、山莨菪碱组,每组10只.方法:采用"结扎双侧椎动脉和夹闭双侧颈总动脉+体循环低血压"建立全脑缺血再灌流模型,缺血20 min,再灌流2 h.山莨菪碱组于再灌流前1 min由股静脉给予山莨菪碱,剂量为10 mg/kg,并以5 mg/h速度维持治疗2 h.缺血再灌流组再灌流期间给予等量生理盐水治疗;假手术组只接受手术,但不结扎和夹闭血管.①采用氧化电极法测定脑线粒体呼吸功能,包括呼吸控制率,磷氧比,氧化磷酸化效率.②采用氧电极法测定脑线粒体内还原型烟酰胺腺嘌呤二核苷酸氧化酶,琥珀酸氧化酶,细胞色素C氧化酶活性.③采用原子吸收光谱法测定脑线粒体钙含量.④采用改良八木国夫法测定脑线粒体丙二醛含量.主要观察指标:①各组家兔大脑皮质线粒体呼吸功能、呼吸链氧化酶活性变化.②各组家兔大脑皮质线粒体内钙和丙二醛含量.结果:30只家兔均进入结果分析.①各组家兔大脑皮质线粒体呼吸控制率、磷氧比、氧化磷酸化效率:缺血再灌流组和山莨菪碱组低于假手术组[烟酰胺腺嘌呤二核苷酸链:呼吸控制率:2.34±0.18,3.58±0.29,4.07±0.38,P＜0.05～0.01;磷氧比:1.77±0.10,2.23±0.14,2.41±0.17,P＜0.05～0.01;氧化磷酸化效率:(527±0.78),(8.03±1.30),(9.63±1.50)μkat/g,P＜0.05～0.01;黄素腺嘌呤二核苷酸链:呼吸控制率:1.47±0.23,2.53±0.28,2.84±0.36,P＜0.05～0.01;磷氧比:0.88±0.09,1.58±0.11,1.73±0.17,P＜0.05～0.01;氧化磷酸化效率:(6.05±1.13),(7.47±1.40),(8.62±1.60)μkat/g,P＜0.05～0.01],山莨菪碱组明显高于缺血再灌注组(P＜0.01).②各组家兔大脑皮质还原型烟酰胺腺嘌呤二核苷酸氧化酶、琥珀酸氧化酶、细胞色素C氧化酶活性:缺血再灌流组和山莨菪碱组低于假手术组[还原型烟酰胺腺嘌呤二核苷酸氧化酶:(2.62±0.35),(4.55±0.48),(5.07±0.60)μkat/g;琥珀酸氧化酶:(1.48±0.17),(1.83±0.22),(2.10±0.28)μkat/g;细胞色素C氧化酶:(5.03±1.12),(7.62±1.23),(9.00±1.53)μkat/g,(P＜0.05～0.01)],山莨菪碱组明显高于缺血再灌注组(P＜0.01).③各组大鼠脑线粒体钙和丙二醛含量:缺血再灌流组和山莨菪碱组高于假手术组[钙:(2.36±0.23),(1.39±0.17),(1.22±0.12)mg/g;丙二醛:[(36.38±10.42),(22.69±9.56),(19.74±7.26)μmol/g,(P＜0.05～0.01)].山莨菪碱组低于缺血再灌流组(P＜0.01).结论:山莨菪碱可通过钙拮抗、抑制脂质过氧化、稳定线粒体膜,减轻线粒体结构损伤,并促进缺血再灌流后线粒体呼吸功能、呼吸链酶活性的恢复,从线粒体水平发挥脑保护作用.
揹景:山莨菪堿是從茄科唐古特莨菪中提取的一種生物堿,是一種良好的細胞保護劑.而線粒體功能變化是反映細胞損傷的最敏感指標之一.目的:觀察山莨菪堿對傢兔全腦缺血再灌流後腦線粒體損傷的影響,探討山莨菪堿的腦保護作用.設計:完全隨機對照實驗.單位:華中科技大學同濟醫學院附屬同濟醫院.材料:實驗于2002-09/12在華中科技大學同濟醫學院附屬同濟醫院急診科實驗室完成.選擇30隻健康傢兔,隨機分為假手術組、缺血再灌流組、山莨菪堿組,每組10隻.方法:採用"結扎雙側椎動脈和夾閉雙側頸總動脈+體循環低血壓"建立全腦缺血再灌流模型,缺血20 min,再灌流2 h.山莨菪堿組于再灌流前1 min由股靜脈給予山莨菪堿,劑量為10 mg/kg,併以5 mg/h速度維持治療2 h.缺血再灌流組再灌流期間給予等量生理鹽水治療;假手術組隻接受手術,但不結扎和夾閉血管.①採用氧化電極法測定腦線粒體呼吸功能,包括呼吸控製率,燐氧比,氧化燐痠化效率.②採用氧電極法測定腦線粒體內還原型煙酰胺腺嘌呤二覈苷痠氧化酶,琥珀痠氧化酶,細胞色素C氧化酶活性.③採用原子吸收光譜法測定腦線粒體鈣含量.④採用改良八木國伕法測定腦線粒體丙二醛含量.主要觀察指標:①各組傢兔大腦皮質線粒體呼吸功能、呼吸鏈氧化酶活性變化.②各組傢兔大腦皮質線粒體內鈣和丙二醛含量.結果:30隻傢兔均進入結果分析.①各組傢兔大腦皮質線粒體呼吸控製率、燐氧比、氧化燐痠化效率:缺血再灌流組和山莨菪堿組低于假手術組[煙酰胺腺嘌呤二覈苷痠鏈:呼吸控製率:2.34±0.18,3.58±0.29,4.07±0.38,P＜0.05～0.01;燐氧比:1.77±0.10,2.23±0.14,2.41±0.17,P＜0.05～0.01;氧化燐痠化效率:(527±0.78),(8.03±1.30),(9.63±1.50)μkat/g,P＜0.05～0.01;黃素腺嘌呤二覈苷痠鏈:呼吸控製率:1.47±0.23,2.53±0.28,2.84±0.36,P＜0.05～0.01;燐氧比:0.88±0.09,1.58±0.11,1.73±0.17,P＜0.05～0.01;氧化燐痠化效率:(6.05±1.13),(7.47±1.40),(8.62±1.60)μkat/g,P＜0.05～0.01],山莨菪堿組明顯高于缺血再灌註組(P＜0.01).②各組傢兔大腦皮質還原型煙酰胺腺嘌呤二覈苷痠氧化酶、琥珀痠氧化酶、細胞色素C氧化酶活性:缺血再灌流組和山莨菪堿組低于假手術組[還原型煙酰胺腺嘌呤二覈苷痠氧化酶:(2.62±0.35),(4.55±0.48),(5.07±0.60)μkat/g;琥珀痠氧化酶:(1.48±0.17),(1.83±0.22),(2.10±0.28)μkat/g;細胞色素C氧化酶:(5.03±1.12),(7.62±1.23),(9.00±1.53)μkat/g,(P＜0.05～0.01)],山莨菪堿組明顯高于缺血再灌註組(P＜0.01).③各組大鼠腦線粒體鈣和丙二醛含量:缺血再灌流組和山莨菪堿組高于假手術組[鈣:(2.36±0.23),(1.39±0.17),(1.22±0.12)mg/g;丙二醛:[(36.38±10.42),(22.69±9.56),(19.74±7.26)μmol/g,(P＜0.05～0.01)].山莨菪堿組低于缺血再灌流組(P＜0.01).結論:山莨菪堿可通過鈣拮抗、抑製脂質過氧化、穩定線粒體膜,減輕線粒體結構損傷,併促進缺血再灌流後線粒體呼吸功能、呼吸鏈酶活性的恢複,從線粒體水平髮揮腦保護作用.
배경:산랑탕감시종가과당고특랑탕중제취적일충생물감,시일충량호적세포보호제.이선립체공능변화시반영세포손상적최민감지표지일.목적:관찰산랑탕감대가토전뇌결혈재관류후뇌선립체손상적영향,탐토산랑탕감적뇌보호작용.설계:완전수궤대조실험.단위:화중과기대학동제의학원부속동제의원.재료:실험우2002-09/12재화중과기대학동제의학원부속동제의원급진과실험실완성.선택30지건강가토,수궤분위가수술조、결혈재관류조、산랑탕감조,매조10지.방법:채용"결찰쌍측추동맥화협폐쌍측경총동맥+체순배저혈압"건립전뇌결혈재관류모형,결혈20 min,재관류2 h.산랑탕감조우재관류전1 min유고정맥급여산랑탕감,제량위10 mg/kg,병이5 mg/h속도유지치료2 h.결혈재관류조재관류기간급여등량생리염수치료;가수술조지접수수술,단불결찰화협폐혈관.①채용양화전겁법측정뇌선립체호흡공능,포괄호흡공제솔,린양비,양화린산화효솔.②채용양전겁법측정뇌선립체내환원형연선알선표령이핵감산양화매,호박산양화매,세포색소C양화매활성.③채용원자흡수광보법측정뇌선립체개함량.④채용개량팔목국부법측정뇌선립체병이철함량.주요관찰지표:①각조가토대뇌피질선립체호흡공능、호흡련양화매활성변화.②각조가토대뇌피질선립체내개화병이철함량.결과:30지가토균진입결과분석.①각조가토대뇌피질선립체호흡공제솔、린양비、양화린산화효솔:결혈재관류조화산랑탕감조저우가수술조[연선알선표령이핵감산련:호흡공제솔:2.34±0.18,3.58±0.29,4.07±0.38,P＜0.05～0.01;린양비:1.77±0.10,2.23±0.14,2.41±0.17,P＜0.05～0.01;양화린산화효솔:(527±0.78),(8.03±1.30),(9.63±1.50)μkat/g,P＜0.05～0.01;황소선표령이핵감산련:호흡공제솔:1.47±0.23,2.53±0.28,2.84±0.36,P＜0.05～0.01;린양비:0.88±0.09,1.58±0.11,1.73±0.17,P＜0.05～0.01;양화린산화효솔:(6.05±1.13),(7.47±1.40),(8.62±1.60)μkat/g,P＜0.05～0.01],산랑탕감조명현고우결혈재관주조(P＜0.01).②각조가토대뇌피질환원형연선알선표령이핵감산양화매、호박산양화매、세포색소C양화매활성:결혈재관류조화산랑탕감조저우가수술조[환원형연선알선표령이핵감산양화매:(2.62±0.35),(4.55±0.48),(5.07±0.60)μkat/g;호박산양화매:(1.48±0.17),(1.83±0.22),(2.10±0.28)μkat/g;세포색소C양화매:(5.03±1.12),(7.62±1.23),(9.00±1.53)μkat/g,(P＜0.05～0.01)],산랑탕감조명현고우결혈재관주조(P＜0.01).③각조대서뇌선립체개화병이철함량:결혈재관류조화산랑탕감조고우가수술조[개:(2.36±0.23),(1.39±0.17),(1.22±0.12)mg/g;병이철:[(36.38±10.42),(22.69±9.56),(19.74±7.26)μmol/g,(P＜0.05～0.01)].산랑탕감조저우결혈재관류조(P＜0.01).결론:산랑탕감가통과개길항、억제지질과양화、은정선립체막,감경선립체결구손상,병촉진결혈재관류후선립체호흡공능、호흡련매활성적회복,종선립체수평발휘뇌보호작용.
BACKGROUND: An isodamine, a kind of alkaloid, is extracted from Anisodus tanguticus (Maxim.) Pascher and is also a good protective agent of cell. However, functional change of mitochondrion is the most sensitive index reflecting cell injury.OBJECTIVE: To study the effects of anisodamine on brain mitochondrial damage following global cerebral ischemia and reperfusion in domestic rabbits and explore its mechanism.DESIGN: Totally randomized controlled trials.SETTING: Emergency Department of Tongji Hospital, Tongji Medical College of Huazhong University of Science and Technology.MATERIALS: The experiment was carried out in the laboratory of Emergency Department, Tongji Hospital Affiliated to Tongji Medical College of Huazhong University of Science and Technology, from September to December in 2002. Thirty healthy domestic rabbits of either sex were used and randomized into sham-operation group, ischemia-reperfusion group and anisodamine group with 10 rabbits in each group.METHODS: The models of complete cerebral ischemia and reperfusion injury in rabbits were established by ligation of bilateral common carotids and vertebral arteries with systemic hypotension, ischemia lasting for 20 minutes followed by 2-hour reperfusion. Anisodamine group was injected with anisodamine at a dose of 10 mg/kg body mass via femoral vein one minute before reperfusion, and lasted for 2 hours at a dose of 5 mg/hour by micro-pump. Ischemia-reperfusion group was treated with normal saline of the same volume. Sham-operation group only underwent used to determine mitochondrial respiratory functions, including respiratory control rate (RCR), the ratio of adenosine diphosphate to oxygen nicotinamide adenine dinucleotide hydrogenated (NADH) oxidase, succinate oxidase and cytochrome C oxidase were measured by the oxygenmethod of Yagi.drial calcium (Ca2+) and malondiadhyde (MDA) in cortex.reperfusion group and anisodamine group, RCR, ADP/O, OPR levels were lower than those in sham-operation group [nicotinamide adenine dinucleotide chain: RCR: 2.34±0.18,3.58±0.29,4.07±0.38,P ＜ 0.05-0.01;ADP/O: 1.77±0.10,2.23±0.14,2.41±0.17,P ＜ 0.05-0.01; OPR: (5.27±0.78),(8.03±1.30), (9.63±1.50)μkat/g, P ＜ 0.05-0.01; flavin adenine dinucleotide chain: RCR: 1.47±0.23,2.53±0.28,2.84±0.36,P ＜ 0.05-0.01;ADP/O: 0.88±0.09,1.58±0.11,1.73±0.17 ,P ＜ 0.05-0.01; OPR: (6.05±1.13),(7.47±1.40), (8.62±1.60)μkat/g,P ＜ 0.05-0.01], and those were higher in chemia-reperfusion group and anisodamine group, the activities of respiratory chain oxidase of NADH, succinate and cytochrome C were lower than those in sham-operation group [NADH: (2.62±0.35), (4.55±0.48), (5.07±0.60)μkat/g;succinate: (1.48±0.17), (1.83±0.22), (2.10±0.28)μkat/g; cytochrome C:(5.03±1.12), (7.62±1.23), (9.00±1.53)μkat/g, P ＜ 0.05-0.01], and those were higher in anisodamine group than in ischemia-reperfusion group, the content of mitochondrial Ca2+ [(2.36±0.23), (1.39±0.17),(1.22±0.12) mg/g] and MDA [(36.38±10.42), (22.69±9.56), (19.74±7.26)μmol/g,(P ＜ 0.05-0.01 )] was higher than that in sham-operation group, and it was lower in anisodamine group than in ischemia-reperfusion group (P ＜ 0.01).CONCLUSION: Anisodamine can protect the brain against ischemiareperfusion injury at the level of mitochondria by antagonism of Ca2+, inhibition of lipid peroxidation, stabilization of mitochondrial membrane, alleviation of mitochondrial damage, and improvement of motochondrial respiratory functions and the activities of enzymes of respiratory chain.
