中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2010年
7期
907-910
,共4页
卢根林%闫兴军%邓勇%斯伟宏%毛樟林%陈小英
盧根林%閆興軍%鄧勇%斯偉宏%毛樟林%陳小英
로근림%염흥군%산용%사위굉%모장림%진소영
肠缺血%再灌注损伤%硫化氢%肝脏功能%脱噬作用
腸缺血%再灌註損傷%硫化氫%肝髒功能%脫噬作用
장결혈%재관주손상%류화경%간장공능%탈서작용
Ischemia of intestine%Reperfusion injury%Hydrogen sulfide%liver function%Apoptosis
目的 观察硫化氢(H2S)在肠缺血-再灌注损伤大鼠肝脏功能障碍中的作用.方法 将24只雄性Wistar大鼠随机分为A(假手术)组、B(缺血-再灌注)组、C(缺血-再灌注+NABS)组(n=8).留取血浆测定H2S、谷丙转氨酶(ALT)、谷草转氨酶(AST)、总胆红素(TBIL),行肝脏组织匀浆检测脂质过氧化物(LPO)、髓过氧化物酶(MPO)、超氧物歧化酶(SOD)、丙二醇(MDA)和GSH/GSSG,光镜和电镜下观察肝脏组织病理变化,TUNEL染色观察肝细胞凋亡指数(AJ),逆转录.聚合酶链反应(RT-PCR)、Western blot法分别检测肝脏组织bcl-2、bax、Caspase-3、STAT3 mRNA和蛋白、pSTAT3和cytochrome C蛋白的表达.结果 C组ALT、AST、肝脏组织MPO、浸润的中性粒细胞数、MDA、LPO、AI、pSTAT3蛋白、Caspase-3mRNA、Caspase-3蛋白、bax mRNA、bax蛋白表达量分别为(47.63±5.04)U/L、(57.63±5.04)U/L、(1.71±0.17)U/mg、(14.13±1.64)个/视野、(23.42±0.69)nmol/mg、(140.13±11.33)nmol/mg、(23.39±1.40)%、0.222±0.019、0.477±0.050、0.214±0.009、0.076±0.004、0.419±0.012,均显著低于B组,高于A组(P<0.01),均与H2S负相关,与AJ正相关.C组H2S、GSH/GSSG、SOD、cytochrome C蛋白、bcl-2 mRNA、bcl-2蛋白表达量分别为(35.27±3.14)μmol/L、9.78±0.56、(90.70±5.69)U/mg、1.132±0.076、0.325±0.052、0.377±0.034,均显著高于B组,低于A组(P<0.01),均与AI负相关,与H2S正相关.pSTAT3与bax、Caspase-3基因表达正相关,与cytochrome C蛋白、bcl-2基因表达负相关.结论 H2s对肠缺血再灌注损伤大鼠肝脏功能障碍起保护作用.
目的 觀察硫化氫(H2S)在腸缺血-再灌註損傷大鼠肝髒功能障礙中的作用.方法 將24隻雄性Wistar大鼠隨機分為A(假手術)組、B(缺血-再灌註)組、C(缺血-再灌註+NABS)組(n=8).留取血漿測定H2S、穀丙轉氨酶(ALT)、穀草轉氨酶(AST)、總膽紅素(TBIL),行肝髒組織勻漿檢測脂質過氧化物(LPO)、髓過氧化物酶(MPO)、超氧物歧化酶(SOD)、丙二醇(MDA)和GSH/GSSG,光鏡和電鏡下觀察肝髒組織病理變化,TUNEL染色觀察肝細胞凋亡指數(AJ),逆轉錄.聚閤酶鏈反應(RT-PCR)、Western blot法分彆檢測肝髒組織bcl-2、bax、Caspase-3、STAT3 mRNA和蛋白、pSTAT3和cytochrome C蛋白的錶達.結果 C組ALT、AST、肝髒組織MPO、浸潤的中性粒細胞數、MDA、LPO、AI、pSTAT3蛋白、Caspase-3mRNA、Caspase-3蛋白、bax mRNA、bax蛋白錶達量分彆為(47.63±5.04)U/L、(57.63±5.04)U/L、(1.71±0.17)U/mg、(14.13±1.64)箇/視野、(23.42±0.69)nmol/mg、(140.13±11.33)nmol/mg、(23.39±1.40)%、0.222±0.019、0.477±0.050、0.214±0.009、0.076±0.004、0.419±0.012,均顯著低于B組,高于A組(P<0.01),均與H2S負相關,與AJ正相關.C組H2S、GSH/GSSG、SOD、cytochrome C蛋白、bcl-2 mRNA、bcl-2蛋白錶達量分彆為(35.27±3.14)μmol/L、9.78±0.56、(90.70±5.69)U/mg、1.132±0.076、0.325±0.052、0.377±0.034,均顯著高于B組,低于A組(P<0.01),均與AI負相關,與H2S正相關.pSTAT3與bax、Caspase-3基因錶達正相關,與cytochrome C蛋白、bcl-2基因錶達負相關.結論 H2s對腸缺血再灌註損傷大鼠肝髒功能障礙起保護作用.
목적 관찰류화경(H2S)재장결혈-재관주손상대서간장공능장애중적작용.방법 장24지웅성Wistar대서수궤분위A(가수술)조、B(결혈-재관주)조、C(결혈-재관주+NABS)조(n=8).류취혈장측정H2S、곡병전안매(ALT)、곡초전안매(AST)、총담홍소(TBIL),행간장조직균장검측지질과양화물(LPO)、수과양화물매(MPO)、초양물기화매(SOD)、병이순(MDA)화GSH/GSSG,광경화전경하관찰간장조직병리변화,TUNEL염색관찰간세포조망지수(AJ),역전록.취합매련반응(RT-PCR)、Western blot법분별검측간장조직bcl-2、bax、Caspase-3、STAT3 mRNA화단백、pSTAT3화cytochrome C단백적표체.결과 C조ALT、AST、간장조직MPO、침윤적중성립세포수、MDA、LPO、AI、pSTAT3단백、Caspase-3mRNA、Caspase-3단백、bax mRNA、bax단백표체량분별위(47.63±5.04)U/L、(57.63±5.04)U/L、(1.71±0.17)U/mg、(14.13±1.64)개/시야、(23.42±0.69)nmol/mg、(140.13±11.33)nmol/mg、(23.39±1.40)%、0.222±0.019、0.477±0.050、0.214±0.009、0.076±0.004、0.419±0.012,균현저저우B조,고우A조(P<0.01),균여H2S부상관,여AJ정상관.C조H2S、GSH/GSSG、SOD、cytochrome C단백、bcl-2 mRNA、bcl-2단백표체량분별위(35.27±3.14)μmol/L、9.78±0.56、(90.70±5.69)U/mg、1.132±0.076、0.325±0.052、0.377±0.034,균현저고우B조,저우A조(P<0.01),균여AI부상관,여H2S정상관.pSTAT3여bax、Caspase-3기인표체정상관,여cytochrome C단백、bcl-2기인표체부상관.결론 H2s대장결혈재관주손상대서간장공능장애기보호작용.
Objective To investigate the role of hydrogen sulfide(H2S) in liver dysfunction during gut ischemia-reperfusion injury in rats. Methods Twenty-four male Wistar rats were randomly divided into three groups(n=8): Group A (sham), B (ischemia-reperfusion), C (ischemia-reperfusion and H2S). Plasma was obtained for the determination of H2S, alanine aminotransferase (ALT) , aspartate aminotrans-ferase (AST) , total bilirubin (TBIL). Hepatic homogenate was gained for the detection of lipid hydroperox-ide (LPO), malondialdehyd (MDA), superoxide dismutase (SOD), GSH/GSSG, myeloperoxidase (MPO). Hepatic morphology was studied by HE staining and transmission electron microscopy. Apoptotic index (AI) was monitored by TUNEL staining. The expression of bcl-2, bax, Caspase-3, STAT3 mRNA was detected by reverse transcription-polymerase chain reaction (RT-PCR). The expression of bcl-2, bax, STAT3, Caspase-3, cytochrome C, pSTAT3 protein was analyzed by Western blotting. Results The contents of ALT, AST, MPO, infiltrating neutrophils, MDA, LPO, AI and the expression of pSTAT3 protein, Caspase-3 mRNA, Caspase-3 protein, bax mRNA, bax protein in group C was (47.63 ±5.04) U/L, (57. 63 ±5.04) U/L, (1.71 ±0.17) U/mg, (14.13 ± 1. 64)/HP, (23.42 ±0.69) nmol/mg, (140.13 ± 11.33) nmol/mg, (23. 39 ± 1.40)% , 0.222 ±0.019, 0.477 ±0.050, 0.214 ±0.009, 0.076 ±0.004, 0.419±0.012 respectively, dramatically lower than in group B, predominantly higher than in group A, which were positively correlated with AI, but negatively with H2S. The contents of HjS, GSH/GSSG, SOD and the expression of cytochrome C protein, bcl-2 mRNA, bcl-2 protein in group C were (35.27 ±3. 14) μnol/L, 9. 78 ±0.56, (90.70 ±5.69) U/mg, 1. 132 ±0.076, 0. 325 ±0.052, 0. 377 ±0.034 respectively , significantly lower than in group A, strikingly higher than in group B, which were negatively correlated with AI, but positively with H2S. The expression of pSTAT3 protein was positively correlated with bax, Caspase-3 gene, but negatively with cytochrome C protein and bcl-2 gene. Conclusion H2S plays a impor-tant role in inhibiting liver dysfunction during gut ischemia-repeifusion injury by attenuating neutrophil infiltration and activation, relieving oxidative stress to hepatocytes, maintaining the antioxidant capacity of hepa-tocytes, down-regulating STAT3 signaling pathway by which bcl-2 gene was down-regulated, Caspase-3 and bax gene up-regulated, the release of cytochrome C from mitochondria to cytoplasm lessened, resulting in less hepatocyte apoptosis.
