HO-1对脓毒症大鼠血浆中HMGB-1表达的影响及其心肌保护机制
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摘要
目的探讨血红素氧合酶-1(HO-1)对脓毒症心肌损伤大鼠血浆中高迁移率族蛋白B-1(HMGB-1)变化的影响及其心肌保护机制。方法采用盲肠结扎穿孔术(CLP)建立脓毒症大鼠模型。SPF级SD雄性大鼠128只,分为4组:假手术组、脓毒症组、钴原卟啉Ⅸ组(CoppⅨ组)、锌原卟啉Ⅸ组(ZnppⅨ组),每组32只。术后分别在0、6、12、24h取全血检测血浆中肌酸激酶同工酶(CK-MB)、心肌肌钙蛋白Ⅰ(cTnⅠ)、心型脂肪酸结合蛋白(H-FABP)及HMGB-1的表达;24h除取全血检测上述指标外,留取心脏组织行苏木素-伊红(HE)染色,观察心肌损伤情况,提取心肌细胞总蛋白,Western Blot检测心肌细胞HMGB-1及HO-1蛋白的表达。结果除假手术组外,各实验组HMGB-1水平随时间增加均升高,其中ZnppⅨ组升高最明显,与脓毒症组相比,差异有统计学意义(P<0.05),与CoppⅨ组相比,差异亦有统计学意义(P<0.01)。光镜下假手术组细胞结构基本正常,CoppⅨ组细胞肿胀、胞核完整,脓毒症组及ZnppⅨ组细胞肿胀明显,核溶解坏死。Western Blot检测结果表明,ZnppⅨ组HMGB-1蛋白表达最高,HO-1蛋白表达最低,而CoppⅨ组HO-1蛋白表达最高,HMGB-1蛋白表达最低。结论 HO-1抑制脓毒症心肌损伤大鼠血浆中HMGB-1的表达,并可能通过抑制HMGB-1的表达而发挥心肌保护作用。
Objective To explore the effect of heme oxygenase-1(HO-1)on the expression of plasma high mobility group protein B1(HMGB-1)in septic rats and its myocardial protection mechanism.Methods The sepsis rat model was established by the cecal ligation and perforation(CLP).One hundred and twentyeight male SD rats of SPF grade were divided into the four groups:the sham operation group,sepsis group,CoppⅨ group and ZnppⅨ group,32 cases in each group.The expression levels of CK-MB,cTnⅠ,H-FABP and HMGB-1 in plasma were measured by collecting the whole blood at postoperative 0,6,12 and 24 h.Besides collecting the whole blood for detecting the above indicators at postoperative 24 h,the cardiac tissue was taken to conduct the HE staining for observing the myocardial injury situation and extracting the total protein of cardiac myocytes.Then the expressions of myocardial cell HMGB-1 and HO-1 protein were detected by Western blot.Results Except for the sham operation group,the level of HMGB-1 in all other groups was increased,among which the ZnppⅨ group increase was most significant,compared with the sepsis group,the difference was statistically significant(P<0.05),compared with the CoppⅨ group,the difference also was statistically significant(P<0.01).Under the light microscope,the cells structure of the sham operation group was basically normal,the cells in the CoppⅨ group were swollen and the nucleus was intact,the cells in the sepsis group and the ZnppⅨgroup were swollen obviously,and the nucleus dissolved and necrosed.The Western blot detection results showed that the expression of HMGB-1 protein was the highest in the ZnppⅨgroup,and the expression of HO-1 protein was the lowest,while the expression of HO-1 protein in the CoppⅨgroup was the highest,and the expression of HMGB-1 protein was the lowest.Conclusion HO-1 inhibits the expression of plasma HMGB-1 in septic rats,moreover may play the myocardial protection role by inhibiting the expression of HMGB-1.
Objective To explore the effect of heme oxygenase-1(HO-1)on the expression of plasma high mobility group protein B1(HMGB-1)in septic rats and its myocardial protection mechanism.Methods The sepsis rat model was established by the cecal ligation and perforation(CLP).One hundred and twentyeight male SD rats of SPF grade were divided into the four groups:the sham operation group,sepsis group,CoppⅨ group and ZnppⅨ group,32 cases in each group.The expression levels of CK-MB,cTnⅠ,H-FABP and HMGB-1 in plasma were measured by collecting the whole blood at postoperative 0,6,12 and 24 h.Besides collecting the whole blood for detecting the above indicators at postoperative 24 h,the cardiac tissue was taken to conduct the HE staining for observing the myocardial injury situation and extracting the total protein of cardiac myocytes.Then the expressions of myocardial cell HMGB-1 and HO-1 protein were detected by Western blot.Results Except for the sham operation group,the level of HMGB-1 in all other groups was increased,among which the ZnppⅨ group increase was most significant,compared with the sepsis group,the difference was statistically significant(P<0.05),compared with the CoppⅨ group,the difference also was statistically significant(P<0.01).Under the light microscope,the cells structure of the sham operation group was basically normal,the cells in the CoppⅨ group were swollen and the nucleus was intact,the cells in the sepsis group and the ZnppⅨgroup were swollen obviously,and the nucleus dissolved and necrosed.The Western blot detection results showed that the expression of HMGB-1 protein was the highest in the ZnppⅨgroup,and the expression of HO-1 protein was the lowest,while the expression of HO-1 protein in the CoppⅨgroup was the highest,and the expression of HMGB-1 protein was the lowest.Conclusion HO-1 inhibits the expression of plasma HMGB-1 in septic rats,moreover may play the myocardial protection role by inhibiting the expression of HMGB-1.
引文
[1]邵义明,姚华国,梁小仲,等.高迁移率族蛋白B-表达水平与大鼠脓毒症严重程度及预后关系的实验研究[J].中国危重病急救医学,2006,18(11):668-672.
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[4]YU Y,YANG Y,BIAN Y,et al.Hydrogen gas protects against intestinal injury in wild type but not NRF2knockout mice with severe sepsis by regulating HO-1 and HMGB-1release[J].Shock,2017,48(3):364-370.
[5]TESSIER J P,THURNER B,JNGLING E,et al.Impairment of glucose metabolism in hearts from rats treated with endotoxin[J].Cardiovasc Res,2003,60(1):119-130.
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[7]ZHAO B,FEI J,CHEN Y,et al.Vitamin C treatment attenuates hemorrhagic shock related multi-organ injuries through the induction of heme oxygenase-1[J].BMCComplement Altern Med,2014,14(1):442.
[8]ZHANG Y,YU J B,LUO X Q,et al.Effect of ERK1/2signaling pathway in electro-acupuncture mediated upregulation of heme oxygenase-1in lungs of rabbits with endotoxic shock[J].Med Sci Monit,2014,20(20):1452-1460.
[9]CHUNG S W,LIU X,MACIAS A A,et al.Heme oxygenase-1-derived Carbon monoxide enhances the host defense response to microbial sepsis in mice[J].J Clin Invest,2008,118(1):239-247.
[10]KIM Y M,KIM H J,CHANG K C.Glycyrrhizin reduces HMGB-1secretion in lipopolysaccharide-activated RAW264.7cells and endotoxemic mice by p38/Nrf2-dependent induction of HO-1[J].Int Immunopharmacol,2015,26(1):112-118.
[2]HUANG L F,YAO Y M,DONG N,et al.Association of high mobility group box-1protein levels with sepsis and outcome of severely burned patients[J].Cytokine,2011,53(1):29-34.
[3]LI Y,XIE K L,CHEN H G,et al.Hydrogen gas inhibits high-mobility group box 1release in septic mice by upregulation of heme oxygenase 1[J].J Surg Res,2015,196(1):136-148.
[4]YU Y,YANG Y,BIAN Y,et al.Hydrogen gas protects against intestinal injury in wild type but not NRF2knockout mice with severe sepsis by regulating HO-1 and HMGB-1release[J].Shock,2017,48(3):364-370.
[5]TESSIER J P,THURNER B,JNGLING E,et al.Impairment of glucose metabolism in hearts from rats treated with endotoxin[J].Cardiovasc Res,2003,60(1):119-130.
[6]LUCKE C,RADEMACHER M,ZIMMERMAN A W,et al.Spin-ystem heterogeneities indicate aSelected-fit mechanism in fatty acid binding to heart-type fatty acid binding protein(HFABP)[J].Biochem J,2001,354(2):259-266.
[7]ZHAO B,FEI J,CHEN Y,et al.Vitamin C treatment attenuates hemorrhagic shock related multi-organ injuries through the induction of heme oxygenase-1[J].BMCComplement Altern Med,2014,14(1):442.
[8]ZHANG Y,YU J B,LUO X Q,et al.Effect of ERK1/2signaling pathway in electro-acupuncture mediated upregulation of heme oxygenase-1in lungs of rabbits with endotoxic shock[J].Med Sci Monit,2014,20(20):1452-1460.
[9]CHUNG S W,LIU X,MACIAS A A,et al.Heme oxygenase-1-derived Carbon monoxide enhances the host defense response to microbial sepsis in mice[J].J Clin Invest,2008,118(1):239-247.
[10]KIM Y M,KIM H J,CHANG K C.Glycyrrhizin reduces HMGB-1secretion in lipopolysaccharide-activated RAW264.7cells and endotoxemic mice by p38/Nrf2-dependent induction of HO-1[J].Int Immunopharmacol,2015,26(1):112-118.