氢饱和生理盐水对重症急性胰腺炎大鼠肺损伤的保护作用及对P38MAPK和NF-κB表达的影响

The protective effect of hydrogen-rich saline on lung injury associated with severe acute pancreatitis and its impact on P38MAPK and NF-κB expression

目的 探讨氢饱和生理盐水对重症急性胰腺炎大鼠肺损伤的保护作用及其对P38MAPK和NF-κB表达的影响.方法 雄性Wistar大鼠54只,随机数字法分为假手术组（SO组）、重症急性胰腺炎组（SAP组）和氢饱和生理盐水处理组（HRS组）,每组大鼠18只.胆胰管逆行注射5％牛磺胆酸钠制备重症急性胰腺炎模型.HRS组在造模成功后5 min尾静脉注射HRS（6 mL/kg）,并皮下HRS补液（20 mL/kg）.SO组、SAP组则在模型成功后5 min经尾静脉注射生理盐水（6 mL/kg）,并皮下生理盐水补液（20 mL/kg）.术后3、12、24 h分批剖杀大鼠,每个时间点6只.分别检测各组大鼠血清淀粉酶（AMY）、脂肪酶（LIP）水平.取新鲜肺组织检测其湿/干比（W/D）.取肺组织病理切片行光镜观察及病理评分.采用免疫组化法检测P38MAPK、p-P38MAPK及NF-κB的表达.结果 HRS组各时间点血清AMY [12 h（5306.7 ±909）vs.（5 435.0 ±441.2）]、LIP [12 h（1 897.8±149.4）vs.（1917.9±106.8）]水平较SAP组相应时间点,差异无统计学意义（P＞0.05）,肺组织W/D [12h （3.12±0.58）vs.（1.87±0.25）]及病理评分[12h （2.14±0.38）vs（3.58±0.32）]较SAP组相应时间点显著降低（P＜0.05）.肺组织12 h时间点P38MAPK的表达在各组间差异无统计学意义;SAP组p-P38MAPK及NF-κB的表达较SO组显著升高,HRS组p-P38MAPK及NF-κB的表达较SAP组显著降低.结论 氢饱和生理盐水对重症急性胰腺炎肺损伤具有保护作用,其机制可能与抗氧化作用以及抑制P38MAPK、NF-κB激活有关.

Objective To investigate the protective effect of hydrogen-rich saline on lung injury associated with severe acute pancreatitis and its impact on P38MAPK and NF-κB expressions.Methods Fifty-four male Wistar rats were randomly （random number） divided into three groups：（1） hydrogen-rich saline treatment group （HRS group,n =18）,in which the rats were treated with hydrogen-rich saline （6 mL/kg） administered intravenously via tail vein and HRS （20 mL/kg） administered subcutaneously at 5 min after successful modeling.（2） Severe acute pancreatitis model group （SAP group,n =18）,in which rats received equivalent volume of normal saline instead of hydrogen-rich saline both intravenously and subcutaneously as in HRS group.（3） Sham operation group （SO group,n =18）,in which rats were treated with sham surgery,and received equivalent volume of normal saline as in SAP group.The model of severe acute pancreatitis （SAP） was made by retrograde injection of 5% sodium taurocholate （1 mL/kg） into cholepancreatic duct.All rats were sacrificed at 3 h,12 h,and 24 h separately after the operation （n =6 at a time）.The levels of serum amylase,lipase were measured.The ratio of wet and dry lung tissues was measured.The histopathological changes of lung tissues were observed under optic microscope.The expressions of P38MAPK,p-P38MAPK and NF-κB were measured by using immunohistochemistry method.Results Compared with SAP group,there were no significant differences in levels of serum amylase [12 h （5306.7±909） vs.（5435.0 ±441.2）] and lipase [12 h （1897.8 ±149.4） vs.（1917.9± 106.8）] in HRS group （P 〉0.05）,but there were significant differences in the ratio of wet and dry lung tissues [12 h （3.12 ± 0.58） vs.（1.87 ± 0.25）] and histopathology scores [12 h （2.14 ± 0.38） vs.（3.58 ±0.32）] （P 〈0.05）.There was no significant difference in expression of P38MAPK in lung tissues among three groups at 12 h.Compared with SO group,the expressions of p-P38MAPK and NF-κB were significant increased in SAP group at 12 h,however,they were lower significantly in HRS group than those in SAP group.Conclusions Hydrogen-rich saline has a protective effect on lung injury associated with severe acute pancreatitis,and its mechanism may be likely related to the antioxidant effect and inhibiting the activation of P38MAPK and NF-κB.