中华肝脏病杂志
中華肝髒病雜誌
중화간장병잡지
CHINESE JOURNAL OF HEPATOLOGY
2012年
7期
537-541
,共5页
张金池%郑国富%吴明祥%吴佳文%欧阳亮远%刘学强
張金池%鄭國富%吳明祥%吳佳文%歐暘亮遠%劉學彊
장금지%정국부%오명상%오가문%구양량원%류학강
肝脏%缺血%再灌注损伤%大鼠%异甘草酸镁
肝髒%缺血%再灌註損傷%大鼠%異甘草痠鎂
간장%결혈%재관주손상%대서%이감초산미
Liver,Ischemia%Reperfusioninjury%Rat%Magnesium isoglycyrrhizinate
目的 探讨异甘草酸镁(MI)对大鼠肢体缺血再灌注肝损伤时磷脂酶A2(PLA2)的影响.方法 选取健康、雄性、清洁级SD大鼠24只,体质量(230±30)g,随机分成3组(每组8只):对照组(C组)、缺血再灌注组(I/R组)、异甘草酸镁治疗组(MI组).C组仅麻醉,肢体没有缺血操作; I/R组于再灌注前颈外静脉注入生理盐水1 ml;MI组于再灌注前同法给异甘草酸镁30mg/kg,计1ml.以橡皮带环绕结扎大鼠左后肢根部至趾掌无血流信号达4h后放开橡皮带,再灌注6h建立大鼠后肢缺血再灌注肝损伤模型.再灌注6h后各组处死动物采集标本,分别取大鼠的血清测定ALT、AST、乳酸脱氢酶(LDH)、肌酸激酶(CK),取肝组织做10％的匀浆,采用硫代巴比妥酸法测匀浆及血清的丙二醛(MDA)、髓过氧化物酶(MPO)值,采用酶联免疫法测匀浆及血清PLA2、肿瘤坏死因子α(TNF α),电镜观察肝组织的超微结构改变.结果 (1)C组肝匀浆PLA2、TNFα、MDA、MPO分别为(74.1± 3.1)μg/g、(152.4±7.9)ng/g、(2.02±0.16)nmol/g、(0.20±0.03)活力单位/g；血清PLA2、TNFα、MDA、MPO分别为(80.3±3.9)μg/L、(121.7± 6.6) ng/L、(4.89±0.64) nmol/ml、(65.62±8.33)活力单位/ml; I/R组肝匀浆PLA2、TNFα、MDA、MPO分别为(94.83±21.99)μg/g、(361.3±46.6)ng/g、(3.038±0.391)nmol/g、(0.422±0.062)活力单位/g;血清PLA2、TNFα、MDA、MPO分别为(118.4±9.3) μg/L、(152.7±15.7) ng/L、(7.30±0.73) nmol/ml、(94.83±21.99)活力单位/ml;MI组肝匀浆PLA2、TNFα、MDA、MFO分别为(84.3±5.8) μg/g、(314.4±13.9)ng/g、(2.49±0.07) nmool/g、(0.31±0.05)活力单位/g ;血清PLA2、TNFα、MDA、MPO分别为(96.0±4.4) μg/L、(137.0±5.5) ng/L、(5.61±0.36) nmol/ml、(74.26±5.81)活力单位/ml,与C组比较,I/R组肝匀浆及血清PLA2、TNFα、MDA、MPO均升高,t值分别为10.743、12.504、5.458、8.939和12.303、5.154、7.019、3.513,P值均＜0.01,差异有统计学意义.与I/R组比较,MI组肝匀浆及血清PLA2、TNFα、MDA、MPO均降低,t值分别为6.170、2.726、3.409、3.946和6.543、2.676、5.926、2.558,P值均＜0.05,差异有统计学意义.(2)C组血清中ALT、AST、CK、LDH分别为(64.0±8.9)U/L、(113.4±18.2) U/L、(286.5±26.5) U/L、(190.8±39.0) U/L;I/R组分别为(381.0±133.8) U/L、(1227.0±383.8)U/L、(3944.9±552.0)U/L、(3050.0±833.3)U/L; MI组分别为(205.6±68.7) U/L、(851.8±126.7)U/L、(1252.5±196.3) U/L、(1177.5±244.0)U/L,与C组比较,I/R组血清中ALT、AST、CK、LDH均明显升高,t值分别为6.687、8.197、19.188、9.376,P值均＜0.01,差异有统计学意义.与I/R组比较,MI组血清中ALT、AST、CK、LDH均明显降低,t值分别为3.298、2.626、12.998、6.100,P值均＜0.05,差异有统计学意义.(3)电镜下组织超微结构I/R组中肝组织损伤明显,而用MI组肝组织的损伤明显减轻. 结论 异甘草酸镁能够通过降低PLA2和TNFα生成而减少炎症介质的释放、抑制氧自由基代谢产物MDA的产生和减少MPO的活性,对肢体缺血再灌注过程中的肝损伤有一定的保护作用.
目的 探討異甘草痠鎂(MI)對大鼠肢體缺血再灌註肝損傷時燐脂酶A2(PLA2)的影響.方法 選取健康、雄性、清潔級SD大鼠24隻,體質量(230±30)g,隨機分成3組(每組8隻):對照組(C組)、缺血再灌註組(I/R組)、異甘草痠鎂治療組(MI組).C組僅痳醉,肢體沒有缺血操作; I/R組于再灌註前頸外靜脈註入生理鹽水1 ml;MI組于再灌註前同法給異甘草痠鎂30mg/kg,計1ml.以橡皮帶環繞結扎大鼠左後肢根部至趾掌無血流信號達4h後放開橡皮帶,再灌註6h建立大鼠後肢缺血再灌註肝損傷模型.再灌註6h後各組處死動物採集標本,分彆取大鼠的血清測定ALT、AST、乳痠脫氫酶(LDH)、肌痠激酶(CK),取肝組織做10％的勻漿,採用硫代巴比妥痠法測勻漿及血清的丙二醛(MDA)、髓過氧化物酶(MPO)值,採用酶聯免疫法測勻漿及血清PLA2、腫瘤壞死因子α(TNF α),電鏡觀察肝組織的超微結構改變.結果 (1)C組肝勻漿PLA2、TNFα、MDA、MPO分彆為(74.1± 3.1)μg/g、(152.4±7.9)ng/g、(2.02±0.16)nmol/g、(0.20±0.03)活力單位/g；血清PLA2、TNFα、MDA、MPO分彆為(80.3±3.9)μg/L、(121.7± 6.6) ng/L、(4.89±0.64) nmol/ml、(65.62±8.33)活力單位/ml; I/R組肝勻漿PLA2、TNFα、MDA、MPO分彆為(94.83±21.99)μg/g、(361.3±46.6)ng/g、(3.038±0.391)nmol/g、(0.422±0.062)活力單位/g;血清PLA2、TNFα、MDA、MPO分彆為(118.4±9.3) μg/L、(152.7±15.7) ng/L、(7.30±0.73) nmol/ml、(94.83±21.99)活力單位/ml;MI組肝勻漿PLA2、TNFα、MDA、MFO分彆為(84.3±5.8) μg/g、(314.4±13.9)ng/g、(2.49±0.07) nmool/g、(0.31±0.05)活力單位/g ;血清PLA2、TNFα、MDA、MPO分彆為(96.0±4.4) μg/L、(137.0±5.5) ng/L、(5.61±0.36) nmol/ml、(74.26±5.81)活力單位/ml,與C組比較,I/R組肝勻漿及血清PLA2、TNFα、MDA、MPO均升高,t值分彆為10.743、12.504、5.458、8.939和12.303、5.154、7.019、3.513,P值均＜0.01,差異有統計學意義.與I/R組比較,MI組肝勻漿及血清PLA2、TNFα、MDA、MPO均降低,t值分彆為6.170、2.726、3.409、3.946和6.543、2.676、5.926、2.558,P值均＜0.05,差異有統計學意義.(2)C組血清中ALT、AST、CK、LDH分彆為(64.0±8.9)U/L、(113.4±18.2) U/L、(286.5±26.5) U/L、(190.8±39.0) U/L;I/R組分彆為(381.0±133.8) U/L、(1227.0±383.8)U/L、(3944.9±552.0)U/L、(3050.0±833.3)U/L; MI組分彆為(205.6±68.7) U/L、(851.8±126.7)U/L、(1252.5±196.3) U/L、(1177.5±244.0)U/L,與C組比較,I/R組血清中ALT、AST、CK、LDH均明顯升高,t值分彆為6.687、8.197、19.188、9.376,P值均＜0.01,差異有統計學意義.與I/R組比較,MI組血清中ALT、AST、CK、LDH均明顯降低,t值分彆為3.298、2.626、12.998、6.100,P值均＜0.05,差異有統計學意義.(3)電鏡下組織超微結構I/R組中肝組織損傷明顯,而用MI組肝組織的損傷明顯減輕. 結論 異甘草痠鎂能夠通過降低PLA2和TNFα生成而減少炎癥介質的釋放、抑製氧自由基代謝產物MDA的產生和減少MPO的活性,對肢體缺血再灌註過程中的肝損傷有一定的保護作用.
목적 탐토이감초산미(MI)대대서지체결혈재관주간손상시린지매A2(PLA2)적영향.방법 선취건강、웅성、청길급SD대서24지,체질량(230±30)g,수궤분성3조(매조8지):대조조(C조)、결혈재관주조(I/R조)、이감초산미치료조(MI조).C조부마취,지체몰유결혈조작; I/R조우재관주전경외정맥주입생리염수1 ml;MI조우재관주전동법급이감초산미30mg/kg,계1ml.이상피대배요결찰대서좌후지근부지지장무혈류신호체4h후방개상피대,재관주6h건립대서후지결혈재관주간손상모형.재관주6h후각조처사동물채집표본,분별취대서적혈청측정ALT、AST、유산탈경매(LDH)、기산격매(CK),취간조직주10％적균장,채용류대파비타산법측균장급혈청적병이철(MDA)、수과양화물매(MPO)치,채용매련면역법측균장급혈청PLA2、종류배사인자α(TNF α),전경관찰간조직적초미결구개변.결과 (1)C조간균장PLA2、TNFα、MDA、MPO분별위(74.1± 3.1)μg/g、(152.4±7.9)ng/g、(2.02±0.16)nmol/g、(0.20±0.03)활력단위/g；혈청PLA2、TNFα、MDA、MPO분별위(80.3±3.9)μg/L、(121.7± 6.6) ng/L、(4.89±0.64) nmol/ml、(65.62±8.33)활력단위/ml; I/R조간균장PLA2、TNFα、MDA、MPO분별위(94.83±21.99)μg/g、(361.3±46.6)ng/g、(3.038±0.391)nmol/g、(0.422±0.062)활력단위/g;혈청PLA2、TNFα、MDA、MPO분별위(118.4±9.3) μg/L、(152.7±15.7) ng/L、(7.30±0.73) nmol/ml、(94.83±21.99)활력단위/ml;MI조간균장PLA2、TNFα、MDA、MFO분별위(84.3±5.8) μg/g、(314.4±13.9)ng/g、(2.49±0.07) nmool/g、(0.31±0.05)활력단위/g ;혈청PLA2、TNFα、MDA、MPO분별위(96.0±4.4) μg/L、(137.0±5.5) ng/L、(5.61±0.36) nmol/ml、(74.26±5.81)활력단위/ml,여C조비교,I/R조간균장급혈청PLA2、TNFα、MDA、MPO균승고,t치분별위10.743、12.504、5.458、8.939화12.303、5.154、7.019、3.513,P치균＜0.01,차이유통계학의의.여I/R조비교,MI조간균장급혈청PLA2、TNFα、MDA、MPO균강저,t치분별위6.170、2.726、3.409、3.946화6.543、2.676、5.926、2.558,P치균＜0.05,차이유통계학의의.(2)C조혈청중ALT、AST、CK、LDH분별위(64.0±8.9)U/L、(113.4±18.2) U/L、(286.5±26.5) U/L、(190.8±39.0) U/L;I/R조분별위(381.0±133.8) U/L、(1227.0±383.8)U/L、(3944.9±552.0)U/L、(3050.0±833.3)U/L; MI조분별위(205.6±68.7) U/L、(851.8±126.7)U/L、(1252.5±196.3) U/L、(1177.5±244.0)U/L,여C조비교,I/R조혈청중ALT、AST、CK、LDH균명현승고,t치분별위6.687、8.197、19.188、9.376,P치균＜0.01,차이유통계학의의.여I/R조비교,MI조혈청중ALT、AST、CK、LDH균명현강저,t치분별위3.298、2.626、12.998、6.100,P치균＜0.05,차이유통계학의의.(3)전경하조직초미결구I/R조중간조직손상명현,이용MI조간조직적손상명현감경. 결론 이감초산미능구통과강저PLA2화TNFα생성이감소염증개질적석방、억제양자유기대사산물MDA적산생화감소MPO적활성,대지체결혈재관주과정중적간손상유일정적보호작용.
Objective To investigate the effects of magnesium isoglycyrrhizinate (MI) on the changes of phospholipase A2 (PLA2) induced during liver tissue injury following limb ischemia/reperfusion (I/R) in rats.Method Twenty-four healthy male Sprague-Dawley rats weighing (230±30)g were randomly divided into three groups (n=8 each) as follows:control (Group C:anesthetization without any ischemia); I/R injury (Group I/R:4 h ischemia induced by rubber band ligation of the left hind limb around the roots of the hind limb,followed by 6 h of reperfusion,with 1 mL normal saline given via tail vein prior to reperfusion); MI-treated group (Group MI:underwent ischemia and reperfusion,with 1 mL MI (30 mg/kg) infused prior to reperfusion).Levels of TNFα and PLA2 in plasma and liver tissue were measured by enzyme-linked immumosorbent assay (ELISA).Levels of plasma alanine aminotransferase (ALT),aspartate aminotransferase (AST),lactate dehydrogenase (LDH),creatine kinase (CK),myeloperoxidase (MPO),and malondialdehyde (MDA),and activities of MPO and MDA in liver tissue were measured by colorimetry.Ultrastructural changes of liver tissue were observed by electron microscopy.Results The MI group had significantly lower PLA2 and TNFα in liver homogenates and serum than the I/R group (both P＜ 0.05).Serum ALT,AST,LDH,and CK were significantly lower in the MI group than in the FR group (all P＜ 0.05),as were the levels of MPO and MDA in liver homogenates and serum (all P＜ 0.05).The I/R group showed significantly more liver tissue damage,which appeared to be attenuated in the MI group.Conclusion MI treatment can inhibit the I/R-induced TNFα,PLA2,and MDA in plasma and liver tissue,as well as decrease the I/R-induced MPO activity in rats.Thus,MI may have protective effects against liver tissue injury following limb ischemia/reperfusion.
