背景:现代药理学研究证实,赤芍总苷具有抑制血小板和红细胞聚集、抗凝和抗血栓、抗动脉粥样硬化、保护心脏和肝脏、抗肿瘤等广泛的药理活性.目的:体外培养制备乳鼠损伤心肌细胞,通过细胞培养液中超氧化物歧化酶活性、丙二醛和一氧化氮含量的变化,分析赤芍总苷对损伤心肌细胞的保护作用.设计:观察对比实验.单位:西安交通大学生命科学与技术学院生物工程系,西安交通大学医学院骨病研究所.材料:实验于2003-02/06在西安交通大学生命科学与技术学院生物工程系完成.选取出生1～3 d的SD乳鼠44只,将培养48 h的心肌细胞制备成42瓶用于实验,随机分为6组:正常对照组、药物损伤组、赤芍总苷0.625 mg/L组、赤芍总苷3.125 mg/L组、赤芍总苷15.625 mg/L组、阳性对照组,7瓶/组.方法:无菌条件下进行心肌细胞原代培养.正常对照组不加任何药物,药物损伤组加入异丙基肾上腺素使其终浓度为100 mg/L,赤芍总苷0.625,3.125,15.625 mg/L组加入异丙基肾上腺素30 min后分别加入浓度为0.625,3.125,15.625 mg/L的赤芍总苷,阳性对照组加入异丙基肾上腺素30min后加入辅酶Q10使其终浓度为100 mg/L.然后进行各项指标检测,超氧化物歧化酶活性采用黄嘌呤氧化酶法测定,丙二醛含量为硫代巴比妥酸法测定,一氧化氮含量用硝酸还原酶法测定.主要观察指标:①各组细胞培养液中超氧化物歧化酶活性的测定.②各组细胞培养液中丙二醛和一氧化氮含量的比较.结果:①各组细胞培养液中超氧化物歧化酶活性的测定:与正常对照组比较,药物损伤组总超氧化物歧化酶、CuZn-超氧化物歧化酶和Mn-超氧化物歧化酶水平均明显下降(P＜0.05或＜0.01);而赤芍总苷各剂量组和阳性对照组上述指标均有不同程度的改善(P＜0.05或＜0.01),其中赤芍总苷15.625 mg/L组的保护作用接近或优于阳性对照组[(79.50±10.67),(80.30±13.50);(48.24±13.26),(49.73±10.23);(31.26±10.22),(30.57±9.57)μkat/L;P＞0.05 ].②各组细胞培养液中丙二醛和一氧化氮含量的比较:药物损伤组明显高于正常对照组(P＜0.01),赤芍总苷各剂量组和阳性对照组均有降低丙二醛和一氧化氮含量的作用,并且赤芍总苷的保护作用呈剂量依赖性,随着赤芍总苷加入量的增高而加强,其中高剂量组的丙二醛含量接近正常对照组水平[(5.41±1.81),(4.48±0.94)μmol/L,P＞0.05],一氧化氮含量与阳性对照组相近[(81.83±9.08),(82.41±12.37)mol/L,P＞0.05].结论:赤芍总苷对异丙基肾上腺素造成的心肌损伤具有保护作用,且呈现剂量依赖关系,可能与增强细胞抗氧化作用、减少自由基和脂质过氧化物导致的细胞膜损伤有关.
揹景:現代藥理學研究證實,赤芍總苷具有抑製血小闆和紅細胞聚集、抗凝和抗血栓、抗動脈粥樣硬化、保護心髒和肝髒、抗腫瘤等廣汎的藥理活性.目的:體外培養製備乳鼠損傷心肌細胞,通過細胞培養液中超氧化物歧化酶活性、丙二醛和一氧化氮含量的變化,分析赤芍總苷對損傷心肌細胞的保護作用.設計:觀察對比實驗.單位:西安交通大學生命科學與技術學院生物工程繫,西安交通大學醫學院骨病研究所.材料:實驗于2003-02/06在西安交通大學生命科學與技術學院生物工程繫完成.選取齣生1～3 d的SD乳鼠44隻,將培養48 h的心肌細胞製備成42瓶用于實驗,隨機分為6組:正常對照組、藥物損傷組、赤芍總苷0.625 mg/L組、赤芍總苷3.125 mg/L組、赤芍總苷15.625 mg/L組、暘性對照組,7瓶/組.方法:無菌條件下進行心肌細胞原代培養.正常對照組不加任何藥物,藥物損傷組加入異丙基腎上腺素使其終濃度為100 mg/L,赤芍總苷0.625,3.125,15.625 mg/L組加入異丙基腎上腺素30 min後分彆加入濃度為0.625,3.125,15.625 mg/L的赤芍總苷,暘性對照組加入異丙基腎上腺素30min後加入輔酶Q10使其終濃度為100 mg/L.然後進行各項指標檢測,超氧化物歧化酶活性採用黃嘌呤氧化酶法測定,丙二醛含量為硫代巴比妥痠法測定,一氧化氮含量用硝痠還原酶法測定.主要觀察指標:①各組細胞培養液中超氧化物歧化酶活性的測定.②各組細胞培養液中丙二醛和一氧化氮含量的比較.結果:①各組細胞培養液中超氧化物歧化酶活性的測定:與正常對照組比較,藥物損傷組總超氧化物歧化酶、CuZn-超氧化物歧化酶和Mn-超氧化物歧化酶水平均明顯下降(P＜0.05或＜0.01);而赤芍總苷各劑量組和暘性對照組上述指標均有不同程度的改善(P＜0.05或＜0.01),其中赤芍總苷15.625 mg/L組的保護作用接近或優于暘性對照組[(79.50±10.67),(80.30±13.50);(48.24±13.26),(49.73±10.23);(31.26±10.22),(30.57±9.57)μkat/L;P＞0.05 ].②各組細胞培養液中丙二醛和一氧化氮含量的比較:藥物損傷組明顯高于正常對照組(P＜0.01),赤芍總苷各劑量組和暘性對照組均有降低丙二醛和一氧化氮含量的作用,併且赤芍總苷的保護作用呈劑量依賴性,隨著赤芍總苷加入量的增高而加彊,其中高劑量組的丙二醛含量接近正常對照組水平[(5.41±1.81),(4.48±0.94)μmol/L,P＞0.05],一氧化氮含量與暘性對照組相近[(81.83±9.08),(82.41±12.37)mol/L,P＞0.05].結論:赤芍總苷對異丙基腎上腺素造成的心肌損傷具有保護作用,且呈現劑量依賴關繫,可能與增彊細胞抗氧化作用、減少自由基和脂質過氧化物導緻的細胞膜損傷有關.
배경:현대약이학연구증실,적작총감구유억제혈소판화홍세포취집、항응화항혈전、항동맥죽양경화、보호심장화간장、항종류등엄범적약리활성.목적:체외배양제비유서손상심기세포,통과세포배양액중초양화물기화매활성、병이철화일양화담함량적변화,분석적작총감대손상심기세포적보호작용.설계:관찰대비실험.단위:서안교통대학생명과학여기술학원생물공정계,서안교통대학의학원골병연구소.재료:실험우2003-02/06재서안교통대학생명과학여기술학원생물공정계완성.선취출생1～3 d적SD유서44지,장배양48 h적심기세포제비성42병용우실험,수궤분위6조:정상대조조、약물손상조、적작총감0.625 mg/L조、적작총감3.125 mg/L조、적작총감15.625 mg/L조、양성대조조,7병/조.방법:무균조건하진행심기세포원대배양.정상대조조불가임하약물,약물손상조가입이병기신상선소사기종농도위100 mg/L,적작총감0.625,3.125,15.625 mg/L조가입이병기신상선소30 min후분별가입농도위0.625,3.125,15.625 mg/L적적작총감,양성대조조가입이병기신상선소30min후가입보매Q10사기종농도위100 mg/L.연후진행각항지표검측,초양화물기화매활성채용황표령양화매법측정,병이철함량위류대파비타산법측정,일양화담함량용초산환원매법측정.주요관찰지표:①각조세포배양액중초양화물기화매활성적측정.②각조세포배양액중병이철화일양화담함량적비교.결과:①각조세포배양액중초양화물기화매활성적측정:여정상대조조비교,약물손상조총초양화물기화매、CuZn-초양화물기화매화Mn-초양화물기화매수평균명현하강(P＜0.05혹＜0.01);이적작총감각제량조화양성대조조상술지표균유불동정도적개선(P＜0.05혹＜0.01),기중적작총감15.625 mg/L조적보호작용접근혹우우양성대조조[(79.50±10.67),(80.30±13.50);(48.24±13.26),(49.73±10.23);(31.26±10.22),(30.57±9.57)μkat/L;P＞0.05 ].②각조세포배양액중병이철화일양화담함량적비교:약물손상조명현고우정상대조조(P＜0.01),적작총감각제량조화양성대조조균유강저병이철화일양화담함량적작용,병차적작총감적보호작용정제량의뢰성,수착적작총감가입량적증고이가강,기중고제량조적병이철함량접근정상대조조수평[(5.41±1.81),(4.48±0.94)μmol/L,P＞0.05],일양화담함량여양성대조조상근[(81.83±9.08),(82.41±12.37)mol/L,P＞0.05].결론:적작총감대이병기신상선소조성적심기손상구유보호작용,차정현제량의뢰관계,가능여증강세포항양화작용、감소자유기화지질과양화물도치적세포막손상유관.
BACKGROUND: It is demonstrated in modern pharmacologic study that total paeony glycoside (TPG) provides extensive pharmacologic activities,such as inhibiting aggregation of platelets and erythrocytes, anticoagulation,antithrombsis, anti-arterial sclerosis, protecting heart and liver, anti-tumor,etc.OBJECTIVE: Neonatal rat cardiomyocytes were cultured in vitro and by the changes of superoxide dismutase (SOD) activity, malondialdehyde (MDA) and nitric oxide (NO) contents in cell culture solution, the protection of TPG on injured cardiomyocytes was analyzed.DESIGN: Controlled observation was designed.SETTING: Bioengineering Department in School of Life Science and Technology of Xi 'an Jiaotong University and Institute of Bone Diseases in Medical School of Xi'an Jiaotong University.MATERIALS: The experiment was performed in Bioengineering Department in School of Life Science and Technology of Xi 'an Jiaotong University from February to June in 2003, in which, 44 SD neonatal rats aged 1-3 days were employed. The 48-hour-cultured cardioryocytes were prepared in 42 bottles and randomized into 6 groups, named normal control (normal group), medicated-injury group (injury group), TPG 0.625 mg/L group,TPG 3.125 mg/L group, TPG 15.625 mg/L group and positive control, 7 bottles in each group.METHODS: Cardiomyocytic primary culture was performed under aseptic condition. No any drug was used in normal group, isoprenaline was added in injury group to terminate the concentration at 100 mg/L, in TPG 0.625, 3.125 and 15.625 mg/L groups, 30 minutes after isoprenaline added, RGP at dosages of 0.625, 3.125 and 15.625 mg/L were added respectively; in positive control, 30 minutes after isoprenaline added, coenzyme Q10 was used to terminate the concentration as 100 mg/L.Afterwards, the assay of every index was performed. Xanthine oxidase (XOD) method was used to assay SOD activity, thiobarbituric acid (TBA)method was to assay MDA content and nitrate reductase (NR) method was to assay NO content.cell culture solution in each group.Compared with normal group, the levels of total SOD, CuZn-SOD and MnSOD were reduced remarkably in injury group (P ＜ 0.05 or P ＜ 0.01).The above-indexes in every TPG group and positive control were improved to different extents (P ＜ 0.05 or ＜ 0.01), in which, the protection of TPG 15.625 mg/L group was near to or superior to positive control [(79.50±10.67), (80.30±13.50); (48.24±13.26), (49.73±10.23); (31.26±10.22),in cell culture solution in each group: Those in injury group were higher remarkably than normal group (P ＜ 0.01). MDA and NO contents were all reduced in every TPG group and positive control and dose dependence presented in TPG protection, the higher the dose was, the stronger the action of TPG on protection was, in which, in high-dose group, MDA content was near to normal group [(5.41±1.81), (4.48±0.94) μmol/L, P ＞ 0.05] and NO content was similar to positive control [(81.83± 9.08), (82.41±12.37) mol/L,P ＞ 0.05].CONCLUSION: TPG protects myocardial injury induced by isoprenaline,indicating dose-dependence relationship, which is probably associated with enhanced anti-oxidation of cell, reduced injury of cellular membrane induced by free radical and lipid oxidant.
