硫化氢及其合成酶抑制剂对小鼠急性胰腺炎自噬功能的影响  

The effects of H2S and its synthetase inhibitor on the autophagy in acute pancreatitis mice

作　　者：徐玲玲[1] 凤辉[1] 殷国建[1] 周春华[1] 王少峰[1] 

机构地区：[1]苏州大学附属第二医院消化科,江苏苏州215000

出　　处：《中华胰腺病杂志》2016年第6期389-393,共5页Chinese Journal of Pancreatology

摘　　要：目的探讨给予外源性硫化氢（H2S）及其合成酶抑制剂炔丙基甘氨酸（PAG）对雨蛙素诱导的小鼠急性胰腺炎（AP）自噬功能的影响。方法60只雄性BALB／c小鼠，按数字表法随机分为对照组、AP组、硫氢化钠（NaHS）组、PAG组。采用连续6次腹腔注射雨蛙素（50μg／kg体重）、每次间隔1h的方法建立小鼠AP模型。NaHS组及PAG组在造模前1h分别腹腔注射NaHS10mg／kg体重、PAG50mg／kg体重，对照组及AP组小鼠腹腔注射等容积生理盐水。于第1次注射雨蛙素12h后处死小鼠，取血检测血清淀粉酶、脂肪酶含量，采用去蛋白分光光度法测定血清H2S含量；取部分胰腺组织常规行病理学检查并进行病理评分；采用Real-timePCR法检测胰腺组织硫化氢合成酶胱硫醚-1裂解酶（CSE）mRNA表达，蛋白质印迹法检测胰腺组织自噬相关的微管相关蛋白轻链3-Ⅱ（LC3-Ⅱ）与LC3-I的比值及泛素结合蛋白（p62）的表达。结果对照组血清淀粉酶、脂肪酶、H2S浓度及胰腺组织CSEmRNA、LC3-Ⅱ／LC3-I值和062表达量分别为（2700±100）U／L、（70±20）U／L、（22．9±1．7）mmol／L、1．0±0．1、0．419±0．080、0．227±0．140；AP组小鼠分别为（17290±500）U／L、（520±40）U／L、（31．3±3．0）mmol／L、5．4±0．4、1．184±0．120、1．985±0．210；NaHS组分另4为（27784±1200）U／L、（900±80）U／L、（38．6±3．3）mmol／L、6．9±0．9、1．600±0．210、4．229±0．050；PAG组分另0为（13750±2000）U／L、（370±20）U／L、（24．5±2．1）mmol／L、4．2±0．5、0．745±0．130、1．203±0．080。AP组上述指标均显著高于对照组，NaHS组又较AP组进一步显著升高，而PAG组较AP组显著降低，差异均有统计学意义（P值均〈0．05）。NaHS组胰腺病理损伤较AP组进一步加重，而PAG组胰腺病理损伤较AP组显著减轻。结论PAG能够降低血清淀粉酶、脂肪酶水平，减轻AP时自噬功Objective To investigate the effect of H2S and its synthetase inhibitor propargylglycine （PAG） on the autophagic function in caerulein-indueed acute pancreatitis （AP） mice. Methods A total of 60 male BALB/c mice were randomly divided into control, AP, Naris and PAG group using random number method. AP was induced in mice via hourly intraperitoneal injection of caerulein （50 μg/kg） continuously for 6 hours. Naris and PAG group received NariS （ 10 mg/kg） or PAG （50 mg/kg） 1 h before the AP induction. A equal volume of normal saline solution was injected in control group and AP group. All the mice were killed at 12 h after the first caerulein injection and blood sample was collected for the detection of serum amylase and lipase content. Deproteinization spectrometry was used to detect serum HE S content, and pancreatic tissue was pathological examined and scored. Real-time PCR detected mRNA expression of CSE, and the protein expression of LC3- Ⅱ/LC3 I and p62 was measured using Western blot. Results Serum amylase, lipase,H2S, CSE mRNA, LC3 Ⅱ/LC3 I and p62 were （2 700 ± 100）U/L, （70 ±20）U/L,（22.9 ± 1.7）mmol/L, 1.0 ± 0.1,0.419 ±0.080, 0.227 ± 0. 140 in control group ; （ 17 290 ±500） U/L, （520±40） U/L, （ 31.3 ± 3.0）mmol/L, 5.4 ± 0.4, 1. 184 ± 0. 120, 1.985 ± 0.210 in AP group; （27 784 ± 1 200）U/L, （900 ± 80）U/L, （38.6 ± 3.3）mmol/L, 6.9 ± 0.9, 1.600 ± 0.210, 4.229 ± 0.050 in NaBS group; （13 750 ± 2 000）U/L, （370 ± 20）U/L, （24.5 ± 2.1）mmol/L, 4.2 ± 0.5, 0.745 ± 0. 130, 1.203 ±0.080 in PAG group. All those biomarkers detected above in AP group significantly increased compared with control group, which were much lower than those in NaBS group, but higher than those in PAG group, and the differences were statistically significant （all P 〈 0.05 ）. Pancreatic histological damage in NaBS group was more severe than that in AP group, which in PAG group was less severe than that in AP group. Conclusions PAG could greatly decreas

关 键 词：胰腺炎 硫化氢 炔丙基甘氨酸 自噬 

分 类 号：R576[医药卫生—消化系统]