中华医学杂志
中華醫學雜誌
중화의학잡지
National Medical Journal of China
2014年
9期
704-708
,共5页
崔华楠%李燕%孙美艳%赵晓勇%吴明营
崔華楠%李燕%孫美豔%趙曉勇%吳明營
최화남%리연%손미염%조효용%오명영
组蛋白类%休克,出血性%炎症%凋亡
組蛋白類%休剋,齣血性%炎癥%凋亡
조단백류%휴극,출혈성%염증%조망
Histones%Shock,hemorrhagic%Inflammation%Apoptosis
目的 探讨晚期炎症因子高迁移率组蛋白1(HMGB1)中和抗体在出血性休克导致小鼠心肌损伤中的保护作用及机制.方法 复制KM小鼠的失血性休克模型,将KM小鼠分为假手术组、对照组、失血性休克模型组、HMGB1中和抗体治疗组(各组n=8).并比较测定血清肌酸激酶(CK)及乳酸脱氢酶(LDH)水平及白细胞介素(IL)-1β、IL-6、肿瘤坏死因子(TNF) -α及HMGB1的浓度.取小鼠的心肌组织固定,做HE染色及电镜观察组织病理变化的分析,免疫组织化学法测定Fas/FasL系统及HMGB1在心肌组织中的表达.逆转录多聚酶链反应实验检测 Bax蛋白、凋亡相关基因(Bcl-2)与Caspase-3蛋白的RNA水平的变化,免疫印迹实验观察心肌中HMGB1、Bax、Bcl-2与Caspase-3的蛋白水平的表达.结果 失血性休克/复苏造成明显的心肌细胞的损伤和凋亡,明显增加了血清中炎症因子的表达,升高了促凋亡因子的mRNA水平的升高,HMGB1中和抗体干预后显著减轻了失血性休克导致的心肌细胞的损伤和凋亡[α-HMGB1干预组与失血性休克组比较(4.5±1.3) 比(8.9±1.9)],减轻了炎症反应[α-HMGB1干预组与失血性休克组比较,IL-1β:(127.7±21.2)比(164.3±30.2);IL-6:(226.7±33.4) 比(402.5±59.5);TNF-α:(100.6±10.7) 比(146.5±15.4)],调节了凋亡相关因子的mRNA水平的变化[α-HMGB1干预组与失血性休克组比较,Bcl-2:(0.25±0.02 )比(0.19±0.02;) Bax:( 0.38±0.04) 比(0.50±0.03);Caspase-3:(0.38±0.04)比(0.56±0.04)].结论 HMGB1中和抗体保护了失血性休克/复苏导致的小鼠心肌细胞的损伤及凋亡,其保护作用可能与其减轻炎症反应及凋亡相关基因的表达有关.
目的 探討晚期炎癥因子高遷移率組蛋白1(HMGB1)中和抗體在齣血性休剋導緻小鼠心肌損傷中的保護作用及機製.方法 複製KM小鼠的失血性休剋模型,將KM小鼠分為假手術組、對照組、失血性休剋模型組、HMGB1中和抗體治療組(各組n=8).併比較測定血清肌痠激酶(CK)及乳痠脫氫酶(LDH)水平及白細胞介素(IL)-1β、IL-6、腫瘤壞死因子(TNF) -α及HMGB1的濃度.取小鼠的心肌組織固定,做HE染色及電鏡觀察組織病理變化的分析,免疫組織化學法測定Fas/FasL繫統及HMGB1在心肌組織中的錶達.逆轉錄多聚酶鏈反應實驗檢測 Bax蛋白、凋亡相關基因(Bcl-2)與Caspase-3蛋白的RNA水平的變化,免疫印跡實驗觀察心肌中HMGB1、Bax、Bcl-2與Caspase-3的蛋白水平的錶達.結果 失血性休剋/複囌造成明顯的心肌細胞的損傷和凋亡,明顯增加瞭血清中炎癥因子的錶達,升高瞭促凋亡因子的mRNA水平的升高,HMGB1中和抗體榦預後顯著減輕瞭失血性休剋導緻的心肌細胞的損傷和凋亡[α-HMGB1榦預組與失血性休剋組比較(4.5±1.3) 比(8.9±1.9)],減輕瞭炎癥反應[α-HMGB1榦預組與失血性休剋組比較,IL-1β:(127.7±21.2)比(164.3±30.2);IL-6:(226.7±33.4) 比(402.5±59.5);TNF-α:(100.6±10.7) 比(146.5±15.4)],調節瞭凋亡相關因子的mRNA水平的變化[α-HMGB1榦預組與失血性休剋組比較,Bcl-2:(0.25±0.02 )比(0.19±0.02;) Bax:( 0.38±0.04) 比(0.50±0.03);Caspase-3:(0.38±0.04)比(0.56±0.04)].結論 HMGB1中和抗體保護瞭失血性休剋/複囌導緻的小鼠心肌細胞的損傷及凋亡,其保護作用可能與其減輕炎癥反應及凋亡相關基因的錶達有關.
목적 탐토만기염증인자고천이솔조단백1(HMGB1)중화항체재출혈성휴극도치소서심기손상중적보호작용급궤제.방법 복제KM소서적실혈성휴극모형,장KM소서분위가수술조、대조조、실혈성휴극모형조、HMGB1중화항체치료조(각조n=8).병비교측정혈청기산격매(CK)급유산탈경매(LDH)수평급백세포개소(IL)-1β、IL-6、종류배사인자(TNF) -α급HMGB1적농도.취소서적심기조직고정,주HE염색급전경관찰조직병리변화적분석,면역조직화학법측정Fas/FasL계통급HMGB1재심기조직중적표체.역전록다취매련반응실험검측 Bax단백、조망상관기인(Bcl-2)여Caspase-3단백적RNA수평적변화,면역인적실험관찰심기중HMGB1、Bax、Bcl-2여Caspase-3적단백수평적표체.결과 실혈성휴극/복소조성명현적심기세포적손상화조망,명현증가료혈청중염증인자적표체,승고료촉조망인자적mRNA수평적승고,HMGB1중화항체간예후현저감경료실혈성휴극도치적심기세포적손상화조망[α-HMGB1간예조여실혈성휴극조비교(4.5±1.3) 비(8.9±1.9)],감경료염증반응[α-HMGB1간예조여실혈성휴극조비교,IL-1β:(127.7±21.2)비(164.3±30.2);IL-6:(226.7±33.4) 비(402.5±59.5);TNF-α:(100.6±10.7) 비(146.5±15.4)],조절료조망상관인자적mRNA수평적변화[α-HMGB1간예조여실혈성휴극조비교,Bcl-2:(0.25±0.02 )비(0.19±0.02;) Bax:( 0.38±0.04) 비(0.50±0.03);Caspase-3:(0.38±0.04)비(0.56±0.04)].결론 HMGB1중화항체보호료실혈성휴극/복소도치적소서심기세포적손상급조망,기보호작용가능여기감경염증반응급조망상관기인적표체유관.
Objective To explore the possible protective effects and mechanisms of neutralizing antibody of HMGB1 on hemorrhagic shock-induced cardiac injury in mice. Methods The KM mice of hemorrhagic shock model were divided into sham (sham), control+IgG, hemorrhagic shock (HS) and HMGB1 neutralizing antibody treatment (HS+α-HMGB1) groups (n=8 each). After modeling, the animals were anesthetized and blood samples collected. The concentrations of creatine kinase (CK) and lactate dehydrogenase (LDH) were measured. And the serum levels of such inflammatory factors as interleukin-1β (IL-1β), interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-α) and HMGB1 were analyzed by enzyme-linked immunosorbent assay (ELISA). The cardiac tissues were harvested, fixed; hemotoxylin-eosin stained and observed under transmission electron microscopy. The histopathological cardiac changes were examined after hemorrhagic shock and resuscitation (HS/R). Immunohistochemical staining was performed to detect the cardiac expressions of HMGB1 and Fas/FasL after HS/R. Reverse transcription polymerase chain reaction (PCR) was performed to analyze the RNA levels of Bax, Bcl-2 and Caspase-3 in cardiac tissues. And the protein levels of HMGB1, Bax, Bcl-2 and Caspase-3 in cardiac tissues were tested by Western blot. Results Hemorrhagic shock and resuscitation resulted in significantly cardiac cell damages, enhanced inflammatory factors in sera and up-regulated the expressions of pro-apoptotic genes and proteins in murine hearts. The lowered protein level of Bcl-2 induced by HS/R was reversed by neutralizing HMGB1 antibody treatment. Neutralizing HMGB1 antibody administration obviously attenuated HS/R-induced cardiac damages and apoptosis (HS +α-HMGB1 group vs HS group, 4.5±1.3 vs 8.9±1.9), inhibited inflammatory responses (HS+α-HMGB1 vs HS, IL-1β: 127.7±21.2 vs 164.3±30.2; IL-6: 226.7±33.4 vs 402.5±59.5; TNF-α:100.6±10.7 vs 146.5±15.4), and modulated apoptosis-associated genes (HS+α-HMGB1 group vs HS group, Bcl-2: 0.25±0.02 vs 0.19±0.02; Bax: 0.38±0.04 vs 0.50±0.03; Caspase-3: 0.38±0.04 vs 0.56±0.04) and proteins expression (HS+α-HMGB1 group vs HS group, Bcl-2: 0.47±0.04 vs 0.3±0.03; Bax: 0.11±0.02 vs 0.17±0.02; Caspase-3: 0.62±0.04 vs 0.8±0.04) in murine hearts after HS/R. Conclusion Neutralizing HMGB1 antibody may protect mice from HS/R-induced cardiac damages and apoptosis. Such an effect is probably due to its anti-inflammatory responses and anti-apoptosis related gene expression.
