饱和氢气生理盐水对盲肠结扎穿孔大鼠肺损伤的干预作用*

doi:10.12047/j.cjap.5767.2019.024

摘要/Abstract

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250~300 g)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10 min腹腔注射生理盐水(10 mg/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10 min腹腔注射生理盐水(10 mg/kg);③Sham+H₂组:手术前10 min腹腔注射饱和氢气生理盐水(10 mg/kg); ④CLP+ H₂组:手术前腹腔注射饱和氢气生理盐水(10 mg/kg)。各组于手术后8 h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250~300 g),随机分为4组(每组8只):①生理盐水组:腹腔注射生理盐水(10 mg/kg);②H₂S组:腹腔注射H₂S供体NaHS(56 μmol/kg);③H₂S+H₂组:腹腔注射NaHS前10 min注射饱和氢气生理盐水(10 mg/kg);④H₂组:腹腔注射生理盐水前10 min注射饱和氢气生理盐水(10 mg/kg)。于给药后8 h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。

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

Abstract: 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, 56 μmol/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-α and 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.

Key words: hydrogen-rich saline, hydrogen sulfide, lung injury, cecal ligation and puncture, rat

翟羽, 周晓红, 刘会, 付洪义, 范亚敏, 黄新莉. 饱和氢气生理盐水对盲肠结扎穿孔大鼠肺损伤的干预作用^*[J]. 中国应用生理学杂志, 2019, 35(2): 107-112.

ZHAI Yu, ZHOU Xiao-hong, LIU Hui, FU Hong-yi, FAN Ya-min, HUANG Xin-li. The interventional effects of saturated hydrogen saline on lung injury in rats with cecal ligation and puncture operation[J]. CJAP, 2019, 35(2): 107-112.

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饱和氢气生理盐水对盲肠结扎穿孔大鼠肺损伤的干预作用^*

The interventional effects of saturated hydrogen saline on lung injury in rats with cecal ligation and puncture operation

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