中华危重病急救医学
中華危重病急救醫學
중화위중병급구의학
Chinese Critical Care Medicine
2015年
6期
498-503
,共6页
张红涛%于泳浩%马小叶%杨涛%胡南%王国林
張紅濤%于泳浩%馬小葉%楊濤%鬍南%王國林
장홍도%우영호%마소협%양도%호남%왕국림
氢气%脓毒症%肠损伤%炎性因子%存活率%小鼠
氫氣%膿毒癥%腸損傷%炎性因子%存活率%小鼠
경기%농독증%장손상%염성인자%존활솔%소서
Hydrogen%Sepsis%Intestinal injury%Inflammatory factor%Survival rate%Mouse
目的：探讨氢气吸入对严重脓毒症小鼠血清炎性因子和肠损伤的影响以及作用机制。方法按随机数字表法将176只雄性ICR小鼠分为4组：假手术组、氢气对照组（假手术+氢气吸入）、模型组（严重脓毒症模型）和氢气治疗组（严重脓毒症模型+氢气吸入），每组44只。采用盲肠结扎穿孔术（CLP）诱导小鼠严重脓毒症模型；氢气吸入为假手术或制模后1h和6h分别吸入2%氢气1h。各组取20只小鼠观察7d存活率；剩余小鼠分别于制模后6、12、24、48 h心脏取血后各处死6只，测定血清肿瘤坏死因子-α（TNF-α）、白细胞介素（IL-6、IL-10）和高迁移率族蛋白B1（HMGB1）水平；光镜下观察小肠组织病理学改变并进行评分，检测肠组织髓过氧化物酶（MPO）和天冬氨酸特异性半胱氨酸蛋白酶3（caspase-3）的活性。结果严重脓毒症小鼠7 d存活率为0；氢气治疗组小鼠7 d存活率提升至60%，与模型组比较差异有统计学意义（P＜0.05）。与假手术组比较，模型组制模后各时间点血清TNF-α、IL-6、IL-10、HMGB1含量均明显升高；肠组织损伤严重，且病理评分较高；肠组织MPO和caspase-3活性也均明显增高（均P＜0.05）。与模型组比较，氢气治疗组血清TNF-α、IL-6和HMGB1含量明显降低〔TNF-α（ng/L）：6 h：110.34±9.28比440.55±25.78，12 h：82.29±8.43比448.36±32.54，24 h：79.68±9.04比346.42±22.24，48 h：80.79±10.06比368.94±31.58；IL-6（ng/L）：12 h：58.68±8.55比158.28±16.73，24 h：46.98±7.58比146.74±18.02，48 h：38.67±8.22比136.45±15.45；HMGB1（μg/L）：6 h：15.75±4.32比55.56±10.04，12 h：32.02±9.33比89.65±15.65，24 h：35.87±8.54比86.44±20.33，48 h：23.85±9.83比98.33±18.88，均P＜0.05〕，血清IL-10含量（ng/L）于制模后24 h和48 h明显升高（24 h：135.44±16.43比79.22±12.03，48 h：110.92±12.54比74.47±11.18，均P＜0.05），肠组织损伤减轻，病理评分（分）明显降低（12 h：1.70±0.06比3.23±0.44，24 h：2.12±0.31比4.51±0.58，48 h：2.03±0.42比4.27±0.58，均P＜0.05），肠组织MPO和caspase-3活性均明显降低〔MPO （U/g）：6 h：13.75±4.21比25.56±5.34，12 h：14.72±4.22比30.53±6.87，24 h：11.62±3.14比33.58±7.24，48 h：11.33±4.03比38.57±8.12；caspase-3（荧光强度）：6 h：0.37±0.07比0.69±0.23，12 h：0.42±0.07比0.86±0.13，24 h：0.53±0.11比1.36±0.23，48 h：0.50±0.08比1.48±0.15，均P＜0.05〕。结论氢气吸入能够降低严重脓毒症小鼠全身炎症反应并减轻由其引起的肠组织损伤，从而改善脓毒症进程，提高存活率。
目的：探討氫氣吸入對嚴重膿毒癥小鼠血清炎性因子和腸損傷的影響以及作用機製。方法按隨機數字錶法將176隻雄性ICR小鼠分為4組：假手術組、氫氣對照組（假手術+氫氣吸入）、模型組（嚴重膿毒癥模型）和氫氣治療組（嚴重膿毒癥模型+氫氣吸入），每組44隻。採用盲腸結扎穿孔術（CLP）誘導小鼠嚴重膿毒癥模型；氫氣吸入為假手術或製模後1h和6h分彆吸入2%氫氣1h。各組取20隻小鼠觀察7d存活率；剩餘小鼠分彆于製模後6、12、24、48 h心髒取血後各處死6隻，測定血清腫瘤壞死因子-α（TNF-α）、白細胞介素（IL-6、IL-10）和高遷移率族蛋白B1（HMGB1）水平；光鏡下觀察小腸組織病理學改變併進行評分，檢測腸組織髓過氧化物酶（MPO）和天鼕氨痠特異性半胱氨痠蛋白酶3（caspase-3）的活性。結果嚴重膿毒癥小鼠7 d存活率為0；氫氣治療組小鼠7 d存活率提升至60%，與模型組比較差異有統計學意義（P＜0.05）。與假手術組比較，模型組製模後各時間點血清TNF-α、IL-6、IL-10、HMGB1含量均明顯升高；腸組織損傷嚴重，且病理評分較高；腸組織MPO和caspase-3活性也均明顯增高（均P＜0.05）。與模型組比較，氫氣治療組血清TNF-α、IL-6和HMGB1含量明顯降低〔TNF-α（ng/L）：6 h：110.34±9.28比440.55±25.78，12 h：82.29±8.43比448.36±32.54，24 h：79.68±9.04比346.42±22.24，48 h：80.79±10.06比368.94±31.58；IL-6（ng/L）：12 h：58.68±8.55比158.28±16.73，24 h：46.98±7.58比146.74±18.02，48 h：38.67±8.22比136.45±15.45；HMGB1（μg/L）：6 h：15.75±4.32比55.56±10.04，12 h：32.02±9.33比89.65±15.65，24 h：35.87±8.54比86.44±20.33，48 h：23.85±9.83比98.33±18.88，均P＜0.05〕，血清IL-10含量（ng/L）于製模後24 h和48 h明顯升高（24 h：135.44±16.43比79.22±12.03，48 h：110.92±12.54比74.47±11.18，均P＜0.05），腸組織損傷減輕，病理評分（分）明顯降低（12 h：1.70±0.06比3.23±0.44，24 h：2.12±0.31比4.51±0.58，48 h：2.03±0.42比4.27±0.58，均P＜0.05），腸組織MPO和caspase-3活性均明顯降低〔MPO （U/g）：6 h：13.75±4.21比25.56±5.34，12 h：14.72±4.22比30.53±6.87，24 h：11.62±3.14比33.58±7.24，48 h：11.33±4.03比38.57±8.12；caspase-3（熒光彊度）：6 h：0.37±0.07比0.69±0.23，12 h：0.42±0.07比0.86±0.13，24 h：0.53±0.11比1.36±0.23，48 h：0.50±0.08比1.48±0.15，均P＜0.05〕。結論氫氣吸入能夠降低嚴重膿毒癥小鼠全身炎癥反應併減輕由其引起的腸組織損傷，從而改善膿毒癥進程，提高存活率。
목적：탐토경기흡입대엄중농독증소서혈청염성인자화장손상적영향이급작용궤제。방법안수궤수자표법장176지웅성ICR소서분위4조：가수술조、경기대조조（가수술+경기흡입）、모형조（엄중농독증모형）화경기치료조（엄중농독증모형+경기흡입），매조44지。채용맹장결찰천공술（CLP）유도소서엄중농독증모형；경기흡입위가수술혹제모후1h화6h분별흡입2%경기1h。각조취20지소서관찰7d존활솔；잉여소서분별우제모후6、12、24、48 h심장취혈후각처사6지，측정혈청종류배사인자-α（TNF-α）、백세포개소（IL-6、IL-10）화고천이솔족단백B1（HMGB1）수평；광경하관찰소장조직병이학개변병진행평분，검측장조직수과양화물매（MPO）화천동안산특이성반광안산단백매3（caspase-3）적활성。결과엄중농독증소서7 d존활솔위0；경기치료조소서7 d존활솔제승지60%，여모형조비교차이유통계학의의（P＜0.05）。여가수술조비교，모형조제모후각시간점혈청TNF-α、IL-6、IL-10、HMGB1함량균명현승고；장조직손상엄중，차병리평분교고；장조직MPO화caspase-3활성야균명현증고（균P＜0.05）。여모형조비교，경기치료조혈청TNF-α、IL-6화HMGB1함량명현강저〔TNF-α（ng/L）：6 h：110.34±9.28비440.55±25.78，12 h：82.29±8.43비448.36±32.54，24 h：79.68±9.04비346.42±22.24，48 h：80.79±10.06비368.94±31.58；IL-6（ng/L）：12 h：58.68±8.55비158.28±16.73，24 h：46.98±7.58비146.74±18.02，48 h：38.67±8.22비136.45±15.45；HMGB1（μg/L）：6 h：15.75±4.32비55.56±10.04，12 h：32.02±9.33비89.65±15.65，24 h：35.87±8.54비86.44±20.33，48 h：23.85±9.83비98.33±18.88，균P＜0.05〕，혈청IL-10함량（ng/L）우제모후24 h화48 h명현승고（24 h：135.44±16.43비79.22±12.03，48 h：110.92±12.54비74.47±11.18，균P＜0.05），장조직손상감경，병리평분（분）명현강저（12 h：1.70±0.06비3.23±0.44，24 h：2.12±0.31비4.51±0.58，48 h：2.03±0.42비4.27±0.58，균P＜0.05），장조직MPO화caspase-3활성균명현강저〔MPO （U/g）：6 h：13.75±4.21비25.56±5.34，12 h：14.72±4.22비30.53±6.87，24 h：11.62±3.14비33.58±7.24，48 h：11.33±4.03비38.57±8.12；caspase-3（형광강도）：6 h：0.37±0.07비0.69±0.23，12 h：0.42±0.07비0.86±0.13，24 h：0.53±0.11비1.36±0.23，48 h：0.50±0.08비1.48±0.15，균P＜0.05〕。결론경기흡입능구강저엄중농독증소서전신염증반응병감경유기인기적장조직손상，종이개선농독증진정，제고존활솔。
Objective To investigate the effects and mechanisms of hydrogen inhalation on serum levels of pro-inflammatory factors and intestinal injury in severe septic mice. Methods 176 male ICR mice were randomly divided into four groups: sham operation group, hydrogen control group ( sham + hydrogen inhalation ), model group ( severe sepsis model ) and hydrogen treatment group ( severe sepsis model+hydrogen inhalation ), with 44 mice in each group. Severe sepsis model was reproduced by cecal ligation and puncture ( CLP ). 2%hydrogen inhalation was given for 1 hour at 1 hour and 6 hours after sham or CLP operation. Twenty animals in each group were selected and observed for 7-day survival rate. Six animals in each group were selected and sacrificed at 6, 12, 24 and 48 hours after sham or CLP, the concentrations of tumor necrosis factor-α ( TNF-α), interleukins ( IL-6, IL-10 ) and high mobility group box 1 ( HMGB1 ) in serum were determined, the intestinal histopathological changes and scores were evaluated by light microscopy, and the activities of myeloperoxidase ( MPO ) and caspase-3 were determined. Results The 7-day survival rate of severe sepsis mice was 0; the 7-day survival rate was increased to 60% in hydrogen treatment group, with statistical significance in variables compared with model group ( P<0.05 ). Compared with sham operation group, the serum concentrations of TNF-α, IL-6, IL-10 and HMGB1 were obviously increased, the intestine were heavily injured along with higher histopathological scores, and the intestinal MPO and caspase-3 activities were significantly enhanced at different time points after CLP in model group ( all P<0.05 ). Compared with model group, the serum concentrations of TNF-α, IL-6 and HMGB1 were significantly decreased [ TNF-α( ng/L ):6 hours:110.34±9.28 vs. 440.55±25.78, 12 hours: 82.29±8.43 vs. 448.36±32.54, 24 hours: 79.68±9.04 vs. 346.42±22.24, 48 hours: 80.79±10.06 vs. 368.94±31.58; IL-6 ( ng/L ): 12 hours: 58.68±8.55 vs. 158.28±16.73, 24 hours: 46.98±7.58 vs. 146.74±18.02, 48 hours: 38.67±8.22 vs. 136.45±15.45; HMGB1 (μg/L ): 6 hours: 15.75±4.32 vs. 55.56±10.04, 12 hours:32.02±9.33 vs. 89.65±15.65, 24 hours: 35.87±8.54 vs. 86.44±20.33, 48 hours: 23.85±9.83 vs. 98.33±18.88, all P<0.05 ], the serum concentrations of IL-10 ( ng/L ) at 24 hours and 48 hours after CLP were obviously increased ( 24 hours:135.44±16.43 vs. 79.22±12.03, 48 hours:110.92±12.54 vs. 74.47±11.18, both P<0.05 ), the intestinal injury were ameliorated with decreased histopathological scores ( 12 hours: 1.70±0.06 vs. 3.23±0.44, 24 hours:2.12±0.31 vs. 4.51±0.58, 48 hours:2.03±0.42 vs. 4.27±0.58, all P<0.05 ), and the intestinal MPO and caspase-3 activities were significantly decreased [ MPO ( U/g ):6 hours:13.75±4.21 vs. 25.56±5.34, 12 hours:14.72±4.22 vs. 30.53±6.87, 24 hours:11.62±3.14 vs. 33.58±7.24, 48 hours:11.33±4.03 vs. 38.57±8.12;caspase-3 ( fluorescence intensity ): 6 hours: 0.37±0.07 vs. 0.69±0.23, 12 hours: 0.42±0.07 vs. 0.86±0.13, 24 hours: 0.53±0.11 vs. 1.36±0.23, 48 hours:0.50±0.08 vs. 1.48±0.15, all P<0.05 ] in hydrogen treatment group. Conclusion Hydrogen inhalation can down-regulate the systemic inflammatory response to ameliorate the intestinal injury, and it may improve the septic process and increase the survival rate of mice with severe sepsis.
