静脉注射含饱和氢气生理盐水对大鼠急性重症胰腺炎的作用及其机制

Experimental studies of effects of hydrogen-rich saline in rats with severe acute pancreatitis

目的探讨静脉注射含饱和氢气生理盐水对大鼠急性重症胰腺炎的作用及其机制。方法采用5％牛磺胆酸钠胰胆管逆行注射方法建立急性重症胰腺炎动物模型，54只sD大鼠用数字随机法随机分成对照组、模型组和治疗组，各18只大鼠。对照组大鼠开腹翻动胰腺后随即关腹，不作其他处理，模型组和治疗组在建模成功后1h经尾静脉分别注射生理盐水或含饱和氢气生理盐水（5ml／kg）。成模24h后处死所有大鼠，检测胰腺组织中丙二醛、髓过氧化物酶（MPO）、超氧化物歧化酶（SOD）及谷胱甘肽的含量，采用酶联免疫吸附试验（ELISA）法检测各组大鼠血清中的肿瘤坏死因子（TNF）-仪，白细胞介素（IL）-6及IL-18水平。反转录（RT）．PCR法检测胰腺组织TNF-αmRNA的表达，免疫组化法检测胰腺组织细胞间黏附分子1（ICAM-1）的表达，并对胰腺行常规病理检查。结果治疗组的胰腺病理评分明显低于模型组（9．7±2．0比13．2±2．5，P〈0．05）。治疗组血清TNF-α、IL-6及IL-18水平均明显低于模型组[（88±17）比（1171±18）pg／ml，（25．2±5．9）比（37．3±4．6）ng／L，（401±100）比（439±103）ng／L，均P〈0．05]。治疗组胰腺组织中丙二醛和MPO的含量均明显低于模型组[（8．2±2．8）比（14．7±2．7）nmol／mg，（0．040±0．叭1）比（0．170±0．071）U／g，均P〈0．05]，而SOD及谷胱甘肽的含量明显高于模型组[（22．1±1．3）比（15．1±1．7）U／mg，（4．8±0．4）比（2．5±0．3）U／mg，均P〈0．05]。治疗组胰腺TNF—αmRNA及ICAM．1的表达均明显低于模型组（0．33±0．17比0．94±0．31，2．0±0．4比2．8±0．8，均P〈0．05）。结论静脉注射含饱和氢气生理盐水能够在一定程度上减轻急性重症胰腺炎病理损害，其机制可能与通过显著抑制急性重症胰腺炎发展主寸稃中的氢化府瀚捅伤反席．减少费忡涕盾的释旃髓减榔彬一陆细群府府右芒．

Objective To explore the mechanism and effectiveness of inducing severe acute pancreatitis （SAP） with saline saturated hydrogen in rats. Methods Based on a random number table, 54 Sprague-Dawley rats were divided into 3 groups： control, SAP and treatment （n = 18 each）. The model of SAP was established by a retrograde injection of 5% sodium taurocholate into biliary -pancreatic duct. The treatment group received an injection of 5 ml/kg hydrogen-rich saline into tail vein at 1 h post-modeling. The control group underwent only pancreatic tipping. All rats were exsanguinated under anesthesia by aortal puncture after 24 hours. The serum levels of tumor necrosis factor-alpha （TNF-α） , interleukin-6 （ IL-6 ） and interleukin-18 （IL-18） in 3 groups were measured by enzyme-linked immunosorbent assay （ELISA）. The expression of TNF-otmRNA in pancreatic tissue was measured by reverse transcription （RT）-PCR. And intercellular adhesion molecule-1 （ ICAM-1 ） in pancreatic tissue was measured by immunohistochemistry. The pancreatic tissues were harvested to examine the microscopic changes. The levels of malonic aldehyde （MDA）, myeloperoxidase （MPO）, superoxide dismutase （SOD） and glutathione （GSH） in pancreatic tissue was measured by specific kits. Results The pathological scores of pancreatic tissue were significantly lower in the treatment group than those in the SAP group（9. 7±2. 0 vs 13.2 ±2. 5, P〈0. 05） . The serum level of TNF-oLwas significantly lower in the treatment group than that in the SAP group （ （ 88 ± 17 ） vs （ 1171±18） pg/ml, P 〈0. 05）. And the serum levels of IL-6 and IL-18 were lower in the treatment group than those in the SAP group （ （25.2 ＋5.9） vs （37. 3 ±4. 6） ng/L, （401±100） vs （439 ±103） ng/L, both P 〈 0.05）. As compared with the SAP group, the levels of MDA and MPO decreased significantly in the treatment group （ （8.2± 2. 8） vs （ 14. 7±2. 7） nmoL/mg, （0. 040±0. 011 ） vs （0. 170±0. 071 ） U/g, both P 〈0. 05 ）. However, the levels of SOD and GSH significantly higher than those of the treatment group （（22.1±1.3） vs （15.1±1.7） U/mg, （4.8±0.4） vs （2.5±0.3） U/mg, both P〈0.05）. The expressions of TNF-α mRNA and ICAM-1 in pancreatic tissues were lower than those of the treatment group （0.33± 0.17 vs 0.94± 0.31, 2.0 ± 0.4 vs 2.8±0.8, both P 〈 0.05） . Conclusion Hydrogen-rich saline can decrease the levels of inflammatory mediators and reduce the pathological damage of pancreas through the inhibition of oxidative stress.