活化沉默信息调节因子1对大鼠严重烫伤早期心肌损害的影响
활화침묵신식조절인자1대대서엄중탕상조기심기손해적영향
Effects of activating silent information regulator 1 on early myocardial damage in severely burned rats
  • 万方数据
    中华烧伤杂志 중화소상잡지
    16
    2014年 3期 194-198 ,共5页

    樊磊%白晓智%杨龙龙%王姝月%杨晨%李超%苏琳琳%吕根法%胡大海

    번뢰%백효지%양룡룡%왕주월%양신%리초%소림림%려근법%호대해

    烧伤%心肌%半胱氨酸天冬氨酸蛋白酶3%白细胞介素1β%肿瘤坏死因子α%沉默信息调节因子1 燒傷%心肌%半胱氨痠天鼕氨痠蛋白酶3%白細胞介素1β%腫瘤壞死因子α%沉默信息調節因子1
    소상%심기%반광안산천동안산단백매3%백세포개소1β%종류배사인자α%침묵신식조절인자1
    Burns%Myocardium%Caspase 3%Interleukin-1 beta%Tumor necrosis factor-alpha%Silent information regulator 1
    目的 探讨活化沉默信息调节因子1(SIRT1)对大鼠严重烫伤早期心肌损害的作用.方法 取24只健康雄性SD大鼠,按照随机数字表法分为假伤组、单纯烫伤组、白藜芦醇治疗组,每组8只.单纯烫伤组和白藜芦醇治疗组大鼠背部于95℃热水浴18s,制成30％ TBSAⅢ度烫伤模型,2组大鼠即刻分别腹腔注射生理盐水10 mL(50 mL/kg)与相同剂量生理盐水+白藜芦醇10 μL(1 g/mL,50 mg/kg);假伤组大鼠仅背部于20℃室温水浴18s模拟烫伤过程.伤后6h收集3组大鼠心脏组织及腹主动脉血,HE染色观察组织形态,ELISA法检测血清肌酸激酶(CK)、乳酸脱氢酶(LDH)含量,蛋白质印迹法和实时荧光定量RT-PCR法检测组织中SIRT1、半胱氨酸天冬氨酸蛋白酶3(caspase-3)的蛋白和SIRT1、caspase-3、IL-1β、TNF-α mRNA表达水平(蛋白水平以灰度值表示).对数据行单因素方差分析、LSD-t检验. 结果 (1)假伤组大鼠心肌纤维呈不规则圆柱形、排列整齐、横纹清晰.单纯烫伤组大鼠心肌间质水肿,心肌纤维排列紊乱并断裂、溶解.白藜芦醇治疗组大鼠心肌间质水肿、心肌纤维排列紊乱及溶解程度均较单纯烫伤组减轻.(2)单纯烫伤组大鼠血清CK、LDH含量分别为(2 385 ±712)、(2 551±196)U/L,显著高于假伤组的(290±59)、(759±60) U/L(￡值分别为9.466、25.452,P值均小于0.01).白藜芦醇治疗组大鼠血清CK、LDH含量[(1 336±149)、(2 209±133)U/L]显著低于单纯烫伤组(￡值分别为-4.506、-4.860,P值均小于0.01).(3)单纯烫伤组大鼠心脏组织SIRT1、caspase-3蛋白表达量分别为0.47±0.11、0.48 ±0.12,显著高于假伤组的0.18±0.06、0.09 ±0.05(t值分别为4.813、9.014,P值均小于0.01).白藜芦醇治疗组大鼠心脏组织SIRT1蛋白表达量(0.74±0.18)显著高于单纯烫伤组(t=4.561,P＜0.01),caspase-3蛋白表达量(0.21 ±0.08)显著低于单纯烫伤组(￡=-6.239,P＜0.01).(4)单纯烫伤组大鼠心脏组织中SIRT1、caspase-3、IL-1β、TNF-α的mRNA表达量分别为2.33 ±0.24、1.96 ±0.20、2.46 ±0.21、1.89±0.37,显著高于假伤组的1.00±0.07、1.00±0.06、1.00±0.08、1.00±0.09(t值分别为14.961、12.823、18.559、6.679,P值均小于0.01).白藜芦醇治疗组大鼠心脏组织中SIRT1的mRNA表达量(2.89±0.31)显著高于单纯烫伤组(t=3.997,P＜0.01),caspase-3、IL-1β、TNF-α的mRNA表达量(1.31±0.08、1.64 ±0.09、1.25 ±0.08)显著低于单纯烫伤组(￡值分别为-8.264、-10.245、-4.818,P值均小于0.01). 结论 SIRT1在大鼠严重烫伤早期的心脏内表达增高,使用白藜芦醇进一步活化SIRT1后,可以减轻心肌组织损伤,减少心肌细胞凋亡,降低IL-1β和TNF-α表达.
    목적 탐토활화침묵신식조절인자1(SIRT1)대대서엄중탕상조기심기손해적작용.방법 취24지건강웅성SD대서,안조수궤수자표법분위가상조、단순탕상조、백려호순치료조,매조8지.단순탕상조화백려호순치료조대서배부우95℃열수욕18s,제성30％ TBSAⅢ도탕상모형,2조대서즉각분별복강주사생리염수10 mL(50 mL/kg)여상동제량생리염수+백려호순10 μL(1 g/mL,50 mg/kg);가상조대서부배부우20℃실온수욕18s모의탕상과정.상후6h수집3조대서심장조직급복주동맥혈,HE염색관찰조직형태,ELISA법검측혈청기산격매(CK)、유산탈경매(LDH)함량,단백질인적법화실시형광정량RT-PCR법검측조직중SIRT1、반광안산천동안산단백매3(caspase-3)적단백화SIRT1、caspase-3、IL-1β、TNF-α mRNA표체수평(단백수평이회도치표시).대수거행단인소방차분석、LSD-t검험. 결과 (1)가상조대서심기섬유정불규칙원주형、배렬정제、횡문청석.단순탕상조대서심기간질수종,심기섬유배렬문란병단렬、용해.백려호순치료조대서심기간질수종、심기섬유배렬문란급용해정도균교단순탕상조감경.(2)단순탕상조대서혈청CK、LDH함량분별위(2 385 ±712)、(2 551±196)U/L,현저고우가상조적(290±59)、(759±60) U/L(￡치분별위9.466、25.452,P치균소우0.01).백려호순치료조대서혈청CK、LDH함량[(1 336±149)、(2 209±133)U/L]현저저우단순탕상조(￡치분별위-4.506、-4.860,P치균소우0.01).(3)단순탕상조대서심장조직SIRT1、caspase-3단백표체량분별위0.47±0.11、0.48 ±0.12,현저고우가상조적0.18±0.06、0.09 ±0.05(t치분별위4.813、9.014,P치균소우0.01).백려호순치료조대서심장조직SIRT1단백표체량(0.74±0.18)현저고우단순탕상조(t=4.561,P＜0.01),caspase-3단백표체량(0.21 ±0.08)현저저우단순탕상조(￡=-6.239,P＜0.01).(4)단순탕상조대서심장조직중SIRT1、caspase-3、IL-1β、TNF-α적mRNA표체량분별위2.33 ±0.24、1.96 ±0.20、2.46 ±0.21、1.89±0.37,현저고우가상조적1.00±0.07、1.00±0.06、1.00±0.08、1.00±0.09(t치분별위14.961、12.823、18.559、6.679,P치균소우0.01).백려호순치료조대서심장조직중SIRT1적mRNA표체량(2.89±0.31)현저고우단순탕상조(t=3.997,P＜0.01),caspase-3、IL-1β、TNF-α적mRNA표체량(1.31±0.08、1.64 ±0.09、1.25 ±0.08)현저저우단순탕상조(￡치분별위-8.264、-10.245、-4.818,P치균소우0.01). 결론 SIRT1재대서엄중탕상조기적심장내표체증고,사용백려호순진일보활화SIRT1후,가이감경심기조직손상,감소심기세포조망,강저IL-1β화TNF-α표체.
    Objective To explore the effects of activating silent information regulator 1 (SIRT1) on early myocardial damage in severely burned rats.Methods Twenty-four healthy male SD rats were divided into sham injury group (SI),scald group (S),and resveratrol (RSV) treatment group (RT) according to the random number table,with 8 rats in each group.Rats in groups S and RT were inflicted with 30％ TBSA full-thickness scald on the back by immersing in 95 ℃ water for 18 s.Immediately after injury,rats in group S were intraperitoneally injected with 10 mL normal saline (50 mL/kg) and those in group RT with 10 mL normal saline (50 mL/kg) + 10 μL RSV in the concentration of 1 g/mL (50 mg/kg).Backs of rats in group SI were immersed in 20 ℃ room temperature water for 18 s to simulate the scald process.Heart tissues and aorta abdominalis blood samples were collected at post injury hour (PIH) 6.The histomorphology of heart tissues was observed with HE staining.The serum contents of creatine kinase (CK) and lactate dehydrogenase (LDH) were determined with ELISA.The protein expressions of SIRT1 and caspase-3 and mRNA expressions of SIRT1,caspase-3,IL-1β,and TNF-α in heart tissue specimens were determined with Western blotting and real-time fluorescent quantitative RT-PCR (with protein level denoted as gray value).Data were processed with one-way analysis of variance and LSD-t test.Results (1) In group SI,myocardial fibers were in irregularly cylindrical shape,neatly arranged,and the transverse striation were distinct.In group S,myocardial interstitial edema,disorder of myocardial fiber arrangement,and cytoplasm destruction were observed.In group RT,the degrees of myocardial interstitial edema,disorder of myocardial fiber arrangement,and cytoplasm destruction were alleviated in comparison with those of group S.(2) The serum contents of CK and LDH of rats in group S were respectively (2 385 ±712) and (2 551 ± 196) U/L,which were significantly higher than those in the group SI [(290 ± 59) and (759 ± 60) U/L,with t values respectively 9.466 and 25.452,P values below 0.01].The serum contents of CK and LDH of rats in group RT were respectively (1 336 ± 149) and (2 209 ± 133) U/L,which were significantly lower than those of group S (with t values respectively-4.506 and-4.860,P values below 0.01).(3) The protein expressions of SIRT1 and caspase-3 in heart tissue of rats in group S were respectively 0.47 ± 0.11 and 0.48 ± 0.12,which were significantly higher than those in group SI [0.18 ± 0.06 and 0.09 ± 0.05,with t values respectively 4.813 and 9.014,P values below 0.01].The protein expression of SIRT1 in heart tissue of rats in group RT was 0.74 ± 0.18,which was significantly higher than that of group S (t =4.561,P ＜ 0.01) ; the protein expression of caspase-3 in heart tissue of rats in group RT was 0.21 ± 0.08,which was significantly lower than that of group S (t =-6.239,P ＜0.01).(4) The mRNA expressions of SIRT1,caspase-3,IL-1β,and TNF-α in heart tissue of rats in group S were respectively 2.33 ± 0.24,1.96 ± 0.20,2.46 ±0.21,1.89 ± 0.37,which were significantly higher than those in group SI (1.00 ± 0.07,1.00±0.06,1.00±0.08,1.00±0.09,witht values respectively 14.961,12.823,18.559,6.679,P values below 0.01).The mRNA expression of SIRT1 in heart tissue of rats in group RT was 2.89 ±0.31,which was significantly higher than that of group S (t =3.997,P ＜ 0.01).The mRNA expressions of caspase-3,IL-1β,and TNF-α in heart tissue of rats in group RT were respectively 1.31 ±0.08,1.64 ± 0.09,1.25 ± 0.08,which were significantly lower than those of group S (with t values respectively -8.264,-10.245,-4.818,Pvalues below0.01).Conclusions The expression of SIRT1 in heart tissue is upregulated in the early stage of severely burned rats.Activation of SIRT1 by RSV can alleviate myocardial tissue injury and reduce apoptosis of cardiac myocytes and secretion of IL-1β and TNF-α.
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