摘要
目的探讨硫化氢(H2S)在重症急性胰腺炎(SAP)中的作用及机制。方法 SD大鼠分为对照组(n=6)、SAP组(n=10)、炔丙基甘氨酸(PAG)+SAP组(n=10)、5mg/kg NaHS+SAP组(n=10)、10mg/kg NaHS+SAP组(n=10)、20mg/kg NaHS+SAP组(n=10)、100mg/kg NaHS+SAP组(n=10)、wortmannin(以下简写W)+SAP组(n=10)、5mg/kg NaHS+W+SAP组(n=10)及100mg/kg NaHS+W+SAP组(n=10),腹腔内注射6%左旋精氨酸制作SD大鼠SAP模型,24h后处死大鼠。ELISA法检测血浆IL-6水平、胰腺髓过氧化物酶(MPO)活性,Western blot法检测各组大鼠胰腺磷酸化丝/苏氨酸蛋白激酶(p-AKT)及核因子抑制蛋白-κB(IκBα)表达水平,EMSA法检测核转录因子-κB(NF-κB)活性。结果 SAP组和PAG+SAP组血浆IL-6水平、胰腺组织MPO活性及p-AKT水平较对照组明显升高(P<0.05),NF-κB活性也得到增强,且PAG+SAP组高于SAP组(P<0.05);而SAP组和PAG+SAP组IκBα水平较对照组明显降低(P<0.05),且PAG+SAP组低于SAP组(P<0.05)。给予不同剂量NaHS后,各组血浆IL-6水平、胰腺组织MPO活性、p-AKT水平及NF-κB活性随着NaHS浓度升高而逐渐降低;IκBα水平却逐渐升高。给予W后,各组血浆IL-6、胰腺MPO活性及p-AKT水平均明显降低,NF-κB活性有一定程度的减弱,而IκBα表达水平明显升高。结论外源性H2S可减轻SAP的炎症程度,且该作用在一定范围内与血浆H2S水平呈正比。H2S通过介导PI3K/AKT-NF-κB通路能减轻SAP胰腺损伤的炎症程度。
Objective To explore the role and mechanism of H2 S in severe acute pancreatitis .Methods SD rats were randomly assigned into several experimental groups :contorl group(n=6) ,SAP group(n=10) ,PAG+SAP group(n=10) ,5 mg/kg NaHS+SAP group(n=10) ,10 mg/kg NaHS+SAP group(n=10) ,20 mg/kg NaHS+SAP group(n=10) ,100 mg/kg NaHS+SAP group (n=10) ,wortmannin(W)+ SAP group(n= 10) ,5 mg/kg NaHS + W+ SAP group(n=10) and 100 mg/kg NaHS+ W+ SAP group(n=10) .These rats were intra-peritoneal injected L-Arginine to get SAP model ,and sacrificed ar 24 h hour after the first in-jection .Plasma IL-6 and MPO activity in pancreas were determined by ELISA .Expression of p-AKT and IκBαin pancreas were de-tected by Western blot ,and NF-κB activity was assessed with EMSA .Results SAP and PAG+ SAP resulted in a significant in-creasing in plasma IL-6 ,MPO activity and expression of p-AKT in pancreas ,when compared with the control group(P〈0 .05) ,and NF-κB activity also increased ,and NF-κB activity in PAG+SAP group was more significant (P〈0 .05);however ,IκBαexpression in SAP and PAG+SAP down-regulated obviously ,when compared with the control group(P〈0 .05) ,and the expression in PAG+SAP group was significantly lower(P〈0 .05) .Different dosage of NaHS reduced that the level of plasma IL-6 ,MPO activity ,ex-pression of p-AKT and NF-κB activity in pancreas changed in different degrees ,yet IκBαexpression in these groups increased gradu-ally .When compared with the SAP group ,5 mg/kg NaHS+SAP and 100 mg/kg NaHS+SAP ,plasma IL-6 ,MPO activity and ex-pression of p-AKT in pancreas significantly depressed after given wortmannin ,besides NF-κB activity also down-regualated;never-theless IκBαexpression up-regulated .Conclusion H2 S in precondition induced the development of an anti-inflammatory activity in SAP ,and in proportion with the concentration of H2 S in blood in a degree .H2 S regulated the degree of SAP injury via PI3K/AKT-NF-κB pathway .
出处
《重庆医学》
CAS
CSCD
北大核心
2014年第29期3890-3894,共5页
Chongqing medicine