摘要
目的探究大黄素(EMO)对重症急性胰腺炎(SAP)大鼠肺泡巨噬细胞(AMs)内质网应激的影响及潜在机制。方法40只大鼠随机分为假手术(SO)组、SAP组、4-苯基丁酸(4-PBA)组和EMO组。利用5%牛磺胆酸钠逆行注射胆胰管建立SAP大鼠模型。HE染色评估胰腺、肺组织病理学改变;计算肺组织湿/干重比;测量动脉血氧分压(PaO_(2))和动脉血二氧化碳分压(PaCO_(2))水平;检测血清淀粉酶活性和炎性因子TNF-α及IL-6水平;TUNEL染色计算肺组织细胞凋亡率;Western blot法检测肺组织中葡萄糖调节蛋白78(GRP78)、活化转录因子6(ATF6)、C/EBP同源蛋白(CHOP)及半胱氨酸蛋白酶-12(Caspase-12)蛋白表达水平。体外培养大鼠AMs(NR8383),分为对照(CON)组、脂多糖(LPS)组、4-PBA组和EMO组。检测细胞中谷胱甘肽(GSH)、丙二醛(MDA)水平;检测细胞中GRP78、ATF6、CHOP及Caspase-12蛋白表达水平。结果与SO组比较,SAP组大鼠胰腺、肺组织病理学评分升高(P<0.05);血清淀粉酶活性、TNF-α及IL-6水平升高(P<0.05);肺组织湿/干重比、细胞凋亡率升高(P<0.05);动脉血PaO_(2)降低且PaCO_(2)升高(P<0.05);肺组织GRP78、ATF6、CHOP及Caspase-12蛋白表达水平升高(P<0.05)。与SAP组比较,4-PBA和EMO组上述参数的改变得到缓解(P<0.05)。体外实验中,与CON组比较,细胞培养上清液中GSH水平降低、MDA水平升高(P<0.05);细胞中GRP78、ATF6、CHOP及Caspase-12蛋白表达水平上调(P<0.05)。与LPS组比较,4-PBA和EMO组上述参数改变得到缓解(P<0.05)。结论大黄素可能通过抑制AMs中ATF6/CHOP通路诱导的氧化应激及细胞凋亡对大鼠SAP相关肺损伤发挥保护作用。
This study was designed to investigate the effect of emodin(EMO)on endoplasmic reticulum stress(ERS)in alveolar macrophages(AMs)of rats with severe acute pancreatitis(SAP)and the underlying mechanism.Forty rats were recruited and randomized into four groups:sham-operation(SO)group,SAP group,4-phenylbutyric acid(4-PBA)group and EMO group.The SAP model was established by retrograde injection of 5%sodium taurocholate into the biliopancreatic duct.HE staining was performed to observe the pathological changes in the pancreas and lung;the wet/dry weight ratio of lung tissue was calculated.Arterial partial pressure of the oxygen(PaO_(2))and carbon dioxide(PaCO_(2))were measured;the serum amylase activity and the levels of inflammatory factors TNF-αand IL-6 were evaluated.TUNEL staining was used to determine the cell apoptosis rate of lung tissue;Western blot was used to detect the protein expression levels of GRP78,ATF6,CHOP,and Caspase-12.Moreover,rat AMs(NR8383)were signed into control(CON)group,lipopolysaccharide(LPS)group,4-phenylbutyric acid(4-PBA)group and EMO group in vitro.Glutathione(GSH)and malondialdehyde(MDA)levels in the cell medium were measured,as were the protein expression levels of GRP78,ATF6,CHOP,and Caspase-12 in AMs.For experiments in vivo,compared with the SO group,the pathological score of pancreatic and lung in SAP rats was increased(P<0.05),the levels of serum amylase activity,IL-6 and TNF-α were increased(P<0.05),the wet/dry weight ratio and apoptosis rate of lung tissue were increased(P<0.05),PaO_(2) was decreased,PaCO_(2) was increased(P<0.05),and the protein expression of GRP78,ATF6,CHOP and Caspase-12 were increased in lung tissue(P<0.05).Compared with the SAP group,these indexes mentioned above in the 4-PBA and EMO groups were reversed(P<0.05).For experiments in vitro,compared with the CON group,GSH levels were decreased,and MDA levels were increased in the cell medium of LPS group(P<0.05),and the expression of GRP78,ATF6,CHOP and Caspase-12 proteins were upregulated(P<0.05).Compared with the LPS group,these indexes mentioned above in in cell medium of the 4-PBA and EMO groups were reversed(P<0.05).In conclusion,the protective effect of EMO on pancreatic and lung injury in SAP rats may be closely related to the inhibition of ATF6/CHOP pathway-induced inflammation and oxidative stress in AMs.
作者
刘欢欢
罗亚岚
葛鹏
张桂信
陈海龙
LIU Huanhuan;LUO Yalan;GE Peng;ZHANG Guixin;CEHN Hailong(Department of Abdominal Emergency Surgery,First Affiliated Hospital,Dalian Medical University,Dalian 116011,China;Institute(College)of Integrative Medicine,Dalian Medical University,Dalian 116044,China;Clinical Laboratory of Integrative Medicine,First Affiliated Hospital,Dalian Medical University,Dalian 116011,China)
出处
《免疫学杂志》
CAS
CSCD
北大核心
2023年第12期1013-1020,共8页
Immunological Journal
基金
国家重点研发计划(2019YFE0119300)
国家自然科学基金(82074158)
关键词
大黄素
重症急性胰腺炎
肺损伤
内质网应激
肺泡巨噬细胞
Emodin
Severe acute pancreatitis
Lung injury
Endoplasmic reticulum stress
Alveolar macrophage
