摘要
目的:观察饱和氢气生理盐水对盲肠结扎穿孔(CLP)大鼠肺组织的作用。方法:将24只健康雄性SD大鼠(体重250~300g)随机分成4组(每组6只):①假手术对照组(Sham组)+生理盐水组:盲肠根部穿过丝线,不行结扎和穿孔,手术前10min腹腔注射生理盐水(10mg/kg);②CLP组+生理盐水组:结扎盲肠根部并用18号针头穿刺2个孔,2个孔相距约1cm,手术前10min腹腔注射生理盐水(10mg/kg);③Sham+H2组:手术前10min腹腔注射饱和氢气生理盐水(10mg/kg);④CLP+H2组:手术前腹腔注射饱和氢气生理盐水(10mg/kg)。各组于手术后8h进行观察:采用生物化学和RT-PCR的方法分别检测大鼠肺组织中CSE/H2S体系的变化。采用H2S供体硫氢化钠(NaHS)诱导的肺组织损伤动物模型,另取32只健康雄性SD大鼠(体重250~300g),随机分为4组(每组8只):①生理盐水组:腹腔注射生理盐水(10mg/kg);②H2S组:腹腔注射H2S供体NaHS(56μmol/kg);③H2S+H2组:腹腔注射NaHS前10min注射饱和氢气生理盐水(10mg/kg);④H2组:腹腔注射生理盐水前10min注射饱和氢气生理盐水(10mg/kg)。于给药后8h测定肺系数,检测肺组织中MDA含量、MPO活性及细胞因子TNF-α、IL-6和IL-10含量,观察肺组织形态学变化。结果:饱和氢气生理盐水可抑制CLP大鼠肺组织中CSE/H2S体系:减少CLP大鼠肺组织中H2S的生成,抑制H2S的合成酶CSE的活性及mRNA表达(P均<0.05);外源性给予H2S(NaHS)可造成肺组织损伤,饱和氢气生理盐水可明显减轻H2S所致的肺组织损伤:大鼠肺系数明显减小(P<0.05),MDA含量下降(P<0.05);肺组织MPO活性下降(P<0.05);大鼠肺间质和肺泡中PMN的浸润程度明显减轻,肺组织形态及IQA接近正常(P<0.05)。结论:饱和氢气生理盐水可通过抑制CLP大鼠肺组织中CSE/H2S体系,发挥其改善CLP大鼠肺组织损伤的作用。
Objective:To investigate the hypothesis that hydrogen could ameliorate cecal ligation and puncture(CLP)-induced lung injury of rats by inhibiting cystathionine-gamma-lyase/hydrogen sulfide(CSE/H2S)system.Methods:A total number of 24 healthy male SD rats weighting 250~300 g were randomly divided into four groups(n=6 in each group):sham operation group(sham group),hydrogen-rich saline control group(H2 group),CLP group and hydrogen-rich saline treatment group(CLP+H2 group).The rats were treated with hydrogen-rich saline or saline 10 min before CLP or sham operation.At 8 h of sham or CLP operation,lung samples were obtained to detect the changes of the CSE/H2S system using biochemical and RT-PCR methods.In order to further confirm the role of H2S during hydrogen improve the lung injury of CLP rats,we also observed the effect of hydrogen-rich saline on the lung injury induced by H2S donor-sodium sodium hydrosulfide(NaHS).Thirty-two healthy male SD rats(250~300 g)were randomly divided into four groups(n=8 in each group):control group,H2S group,H2S+H2 group and H2 group.Saline(10 mg/kg)or NaHS(H2S donor,56μmol/kg)was injected intraperitoneally(10 mg/kg)respectively into rats in the control rats or H2S group.For rats in the H2S+H2 and H2 group,hydrogen-rich saline(10 mg/kg)was injected 10 min before saline or NaHS administration.Eight hours after the LPS saline or NaHS administration,lung coefficient,MDA content,and MPO activity were detected.The contents of TNF-α,IL-6 and IL-10 in lung tissue were measured,and the morphological changes of lung tissue were also observed.Results:CSE/H2S system up-regulating were observed in animals exposed to CLP.Hydrogen-rich saline treatment significantly inhibited CSE/H2S system as indicated by significantly reduced H2S production in lung,along with a decreased CSE activity and CSE mRNA expression(all P<0.05).Importantly,the results showed that lung injury and lung tissue inflammation were observed in animals exposed to NaHS.Hydrogen-rich saline treatment significantly attenuated lung injury as indicated by significantly improved histological changes in lung,significantly reduced index of quantitative assessment(IQA),MDA content and lung coefficient(all P<0.05).MPO activity in lung tissue was significantly reduced along with decreased productions of TNF-αand IL-6,and an increased production of IL-10 in the presence of hydrogen(all P<0.05),demonstrating antioxidant and anti-inflammatory effect of hydrogen in NaHS-induced ALI.Conclusion:These results indicate that hydrogen-rich saline peritoneal injection improves the lung injury induced by CLP operation.The therapeutic effects of hydrogen-rich saline may be related to suppressing the production of H2S.
作者
翟羽
周晓红
刘会
付洪义
范亚敏
黄新莉
ZHAI Yu;ZHOU Xiao-hong;LIU Hui;FU Hong-yi;FAN Ya-min;HUANG Xin-li(Department of Basic Medicine,Hebei University of Chinese Medicine,Shijiazhuang 050200;Department of Tuberculosis and Respiratory Medicine,Chest Hospital of Hebei Province,Shijiazhuang 050041;Department of Pathophysiology,Hebei Medical University,Shijiazhuang 050017,China)
出处
《中国应用生理学杂志》
CAS
CSCD
北大核心
2019年第2期107-113,共7页
Chinese Journal of Applied Physiology
基金
国家自然科学基金资助项目(30800440)
河北省自然科学基金资助项目(C2008001040,H2016206191)
河北省教育厅重点项目(ZD2017050)
关键词
饱和氢气生理盐水
硫化氢
肺损伤
盲肠结扎穿孔
大鼠
hydrogen-rich saline
hydrogen sulfide
lung injury
cecal ligation and puncture
rat
