中华医学杂志
中華醫學雜誌
중화의학잡지
National Medical Journal of China
2013年
12期
939-943
,共5页
冯永强%柴家科%褚万立%段红杰%马丽%张海军
馮永彊%柴傢科%褚萬立%段紅傑%馬麗%張海軍
풍영강%시가과%저만립%단홍걸%마려%장해군
烧伤%心肌%线粒体%离子通道%大鼠
燒傷%心肌%線粒體%離子通道%大鼠
소상%심기%선립체%리자통도%대서
Burns%Myocardium%Mitochondria%Ion channels%Rats
目的 探讨电压依赖负离子通道2(VDAC2)相关的线粒体凋亡途径在严重烧伤大鼠心肌损伤的作用及其调控机制.方法 清洁级Wistar大鼠60只(体重198～219 g),按随机数字表法分为假伤组、烫伤组各30只.伤后第1、7、14天收集大鼠血清及心肌组织.酶联免疫吸附(ELISA)法检测血清内心肌钙蛋白Ⅰ(cTnI)水平,Western印迹法检测总蛋白内VDAC2、Bcl-2、Bax、细胞色素C、磷脂酰肌醇3激酶(PI3K)、磷酸化糖原合成酶激酶3β(p-GSK3β)、己糖激酶2(HK2)的表达水平,检测胞质蛋白内细胞色素C的表达水平.结果 烫伤组与假伤组第1、7、14天cTnI浓度分别为(1.41 ±0.25)、(1.93 ±0.53)、(1.62±0.34) μg/L与(0.53±0.23)、(0.43±0.23)、(0.41±0.22)μg/L,差异均有统计学意义(均P＜0.05)；Bax/Bcl-2比值分别为(3.360±0.173)、(2.736 ±0.341)、(1.290±0.234)与(0.623 ±0.044)、(0.698±0.064)、(0.718±0.063),差异均有统计学意义(均P＜0.05).烫伤组与假伤组第1、7、14天VDAC2表达水平分别为0.070±0.009、0.007±0.002、0.009±0.004和0.328±0.020、0.291±0.025、0.302±0.037,胞质内细胞色素C表达分别为0.418±0.030、1.685±0.169、0.300±0.037和0.022±0.007、0.030 ±0.011、0.098±0.014,差异均有统计学意义(均P ＜0.05).烫伤组与假伤组第14天PI3K表达水平为0.083±0.015与0.328±0.011,差异有统计学意义(P＜0.05).各时相点烫伤组p-GSK3β表达水平分别为0.098±0.014、0.064±0.002、0.074±0.010,均低于假伤组的0.446 ±0.031、0.476±0.054、0.442±0.041,差异均有统计学意义(均P＜0.05).烫伤组第7、14天HK2表达水平分别为0.390±0.027、0.267±0.018,均低于假伤组的0.611±0.070、0.490±0.042,差异均有统计学意义(均P＜0.05).结论 VDAC2相关线粒体凋亡途径加重严重烫伤后的心肌损伤,该过程可能受PI3K-GSK3β-HK2信号通路调控.
目的 探討電壓依賴負離子通道2(VDAC2)相關的線粒體凋亡途徑在嚴重燒傷大鼠心肌損傷的作用及其調控機製.方法 清潔級Wistar大鼠60隻(體重198～219 g),按隨機數字錶法分為假傷組、燙傷組各30隻.傷後第1、7、14天收集大鼠血清及心肌組織.酶聯免疫吸附(ELISA)法檢測血清內心肌鈣蛋白Ⅰ(cTnI)水平,Western印跡法檢測總蛋白內VDAC2、Bcl-2、Bax、細胞色素C、燐脂酰肌醇3激酶(PI3K)、燐痠化糖原閤成酶激酶3β(p-GSK3β)、己糖激酶2(HK2)的錶達水平,檢測胞質蛋白內細胞色素C的錶達水平.結果 燙傷組與假傷組第1、7、14天cTnI濃度分彆為(1.41 ±0.25)、(1.93 ±0.53)、(1.62±0.34) μg/L與(0.53±0.23)、(0.43±0.23)、(0.41±0.22)μg/L,差異均有統計學意義(均P＜0.05)；Bax/Bcl-2比值分彆為(3.360±0.173)、(2.736 ±0.341)、(1.290±0.234)與(0.623 ±0.044)、(0.698±0.064)、(0.718±0.063),差異均有統計學意義(均P＜0.05).燙傷組與假傷組第1、7、14天VDAC2錶達水平分彆為0.070±0.009、0.007±0.002、0.009±0.004和0.328±0.020、0.291±0.025、0.302±0.037,胞質內細胞色素C錶達分彆為0.418±0.030、1.685±0.169、0.300±0.037和0.022±0.007、0.030 ±0.011、0.098±0.014,差異均有統計學意義(均P ＜0.05).燙傷組與假傷組第14天PI3K錶達水平為0.083±0.015與0.328±0.011,差異有統計學意義(P＜0.05).各時相點燙傷組p-GSK3β錶達水平分彆為0.098±0.014、0.064±0.002、0.074±0.010,均低于假傷組的0.446 ±0.031、0.476±0.054、0.442±0.041,差異均有統計學意義(均P＜0.05).燙傷組第7、14天HK2錶達水平分彆為0.390±0.027、0.267±0.018,均低于假傷組的0.611±0.070、0.490±0.042,差異均有統計學意義(均P＜0.05).結論 VDAC2相關線粒體凋亡途徑加重嚴重燙傷後的心肌損傷,該過程可能受PI3K-GSK3β-HK2信號通路調控.
목적 탐토전압의뢰부리자통도2(VDAC2)상관적선립체조망도경재엄중소상대서심기손상적작용급기조공궤제.방법 청길급Wistar대서60지(체중198～219 g),안수궤수자표법분위가상조、탕상조각30지.상후제1、7、14천수집대서혈청급심기조직.매련면역흡부(ELISA)법검측혈청내심기개단백Ⅰ(cTnI)수평,Western인적법검측총단백내VDAC2、Bcl-2、Bax、세포색소C、린지선기순3격매(PI3K)、린산화당원합성매격매3β(p-GSK3β)、기당격매2(HK2)적표체수평,검측포질단백내세포색소C적표체수평.결과 탕상조여가상조제1、7、14천cTnI농도분별위(1.41 ±0.25)、(1.93 ±0.53)、(1.62±0.34) μg/L여(0.53±0.23)、(0.43±0.23)、(0.41±0.22)μg/L,차이균유통계학의의(균P＜0.05)；Bax/Bcl-2비치분별위(3.360±0.173)、(2.736 ±0.341)、(1.290±0.234)여(0.623 ±0.044)、(0.698±0.064)、(0.718±0.063),차이균유통계학의의(균P＜0.05).탕상조여가상조제1、7、14천VDAC2표체수평분별위0.070±0.009、0.007±0.002、0.009±0.004화0.328±0.020、0.291±0.025、0.302±0.037,포질내세포색소C표체분별위0.418±0.030、1.685±0.169、0.300±0.037화0.022±0.007、0.030 ±0.011、0.098±0.014,차이균유통계학의의(균P ＜0.05).탕상조여가상조제14천PI3K표체수평위0.083±0.015여0.328±0.011,차이유통계학의의(P＜0.05).각시상점탕상조p-GSK3β표체수평분별위0.098±0.014、0.064±0.002、0.074±0.010,균저우가상조적0.446 ±0.031、0.476±0.054、0.442±0.041,차이균유통계학의의(균P＜0.05).탕상조제7、14천HK2표체수평분별위0.390±0.027、0.267±0.018,균저우가상조적0.611±0.070、0.490±0.042,차이균유통계학의의(균P＜0.05).결론 VDAC2상관선립체조망도경가중엄중탕상후적심기손상,해과정가능수PI3K-GSK3β-HK2신호통로조공.
Objective To explore the role of voltage dependent anion channel 2 (VDAC2) involved mitochondrial apoptosis in heart injury of rats with severe scald injury and elucidate its possible regulatory signal pathway.Methods A total of 60 Wistar rats were divided into sham scald group (n =30) and scald group (n =30) according to a random digital table.Blood and heart tissue samples were harvested at Day 1,7,14 post scalding.Myocardial injury was assessed with cardiac troponin Ⅰ (cTnI) by enzyme-linked immunosorbent assay (ELISA).Mitochondrial apoptosis activation was evaluated by the expressions of Bax/Bcl-2 ratio,cytoplasmic cytochrome C and VDAC2.And the levels of phosphatidylinositol 3-kinase,p-Glycogen Synthase Kinase-3β and hexokinase 2 protein were determined by Western blot.Results The serum levels of cTnI were significantly higher in scald group than those in sham scald group at Day 1,7,14((1.41 ±0.25) vs (0.53 ±0.23) μg/L,(1.93 ±0.53) vs (0.43 ±0.23) pμg/L,(1.62 ±0.34) vs (0.41 ± 0.22) μg/L respectively,all P ＜ 0.05).Compared with sham scald group,Bax/Bcl-2 ratio increased significantly in scald group at Day 1,7 day post-scalding (3.360 ± 0.173 vs 0.623 ± 0.044,2.736 ± 0.341 vs 0.698 ± 0.064,1.290 ± 0.234 vs 0.718 ± 0.063 respectively,all P ＜ 0.05),VDAC2 protein level in scald group decreased significantly at Day 1,7,14 (0.070 ± 0.009 vs 0.328 ± 0.026,0.007 ±0.002 vs 0.291 ± 0.025,0.009 ± 0.004 vs 0.302 ± 0.037 respectively,all P ＜ 0.05),the cytoplasmic levels of cytochrome increased significantly in scald group at Day 1,7,14 (0.418 ± 0.030 vs 0.022 ±0.007,1.685 ±0.169 vs 0.030 ±0.011,0.300 ±0.037 vs 0.098 ±0.014 respectively,all P ＜0.05),the expression of PI3K was significantly lower in scald group at Day 14 post-scalding (0.083 ± 0.015 vs 0.328 ± 0.011,P ＜ 0.05),the expressions of p-GSK3β all reduced significantly at Day 1,7,14(0.098 ±0.014 vs 0.446 ± 0.031,0.064 ± 0.002 vs 0.476 ± 0.054、0.074 ± 0.010 vs 0.442 ± 0.041,respectively,all P ＜ 0.05) and the expressions of HK2 were lower at Day 7,14 post-scalding (0.390 ±0.027 vs 0.611 ±0.070,0.267 ±0.018 vs 0.490 ±0.042,respectively,all P ＜0.05).Conclusions VDAC2 involved mitochondrial apoptosis is activated in myocardium after severe scalds.And it may be regulated by the pathway of PI3K-GSK-HK2.
