PERK在重症急性胰腺炎大鼠淋巴细胞中的表达及其与细胞凋亡的相关性
|
摘要
目的探讨重症急性胰腺炎(SAP)大鼠淋巴细胞蛋白激酶R样内质网激酶(PERK)表达水平的变化及其与凋亡的相关性。方法成年雄性SPF级SD大鼠20只,随机分为假手术组(SO组)、SAP组,每组10只。SAP组大鼠采用5%牛磺胆酸钠胰胆管逆行注射法建立SAP模型,SO组大鼠仅翻动胰腺后关腹。术后18h取材,HE染色观察胰腺组织病理损伤程度并进行评分,qPCR检测脾脏淋巴细胞PERK mRNA、Caspase-3mRNA的表达情况,ELISA法检测血清中Caspase-3含量。结果与SO组比较,SAP组脾脏淋巴细胞PERK mRNA、Caspase-3mRNA表达明显升高(P<0.05),血清中Caspase-3含量升高(P<0.05),脾淋巴细胞PERK mRNA水平与脾淋巴细胞Caspase-3 mRNA、血清Caspase-3水平呈正相关(P<0.05)。结论 PERK表达升高与SAP淋巴细胞凋亡有关,可作为SAP治疗的靶点。
Objective To investigate the changes of protein kinase R-like endoplasmic reticulum kinase(PERK)expression in lymphocytes of rats with severe acute pancreatitis(SAP)and its correlation with apoptosis.Methods Twenty adult male SPF SD rats were randomly divided into a sham operation group(SO group)and a SAP group,with 10 rats in each group.In the SAP group,the SAP model was established by retrograde injection of 5%sodium taurocholate into the pancreatic duct.We only turned the pancreas in the rats of the SO group and closed the rats' abdomen.Eighteen hours after surgery,the pathological damage of pancreatic tissue was observed by HE staining and scored.The expressions of PERK mRNA and Caspase-3 mRNA in spleen lymphocytes were detected by qPCR.The content of Caspase-3 in serum was detected by ELISA.Results Compared with SO group,the expression of PERK mRNA and Caspase-3 mRNA in spleen lymphocytes of SAP group was significantly increased(P<0.05),the content of Caspase-3 in serum was increased(P<0.05),the level of PERK mRNA in spleen lymphocytes was positively correlated with spleen lymphocytes Caspase-3 mRNA and serum Caspase-3 levels(P<0.05).Conclusion Increased expression of PERK is associated with apoptosis of SAP lymphocytes and may be a target for SAP treatment.
Objective To investigate the changes of protein kinase R-like endoplasmic reticulum kinase(PERK)expression in lymphocytes of rats with severe acute pancreatitis(SAP)and its correlation with apoptosis.Methods Twenty adult male SPF SD rats were randomly divided into a sham operation group(SO group)and a SAP group,with 10 rats in each group.In the SAP group,the SAP model was established by retrograde injection of 5%sodium taurocholate into the pancreatic duct.We only turned the pancreas in the rats of the SO group and closed the rats' abdomen.Eighteen hours after surgery,the pathological damage of pancreatic tissue was observed by HE staining and scored.The expressions of PERK mRNA and Caspase-3 mRNA in spleen lymphocytes were detected by qPCR.The content of Caspase-3 in serum was detected by ELISA.Results Compared with SO group,the expression of PERK mRNA and Caspase-3 mRNA in spleen lymphocytes of SAP group was significantly increased(P<0.05),the content of Caspase-3 in serum was increased(P<0.05),the level of PERK mRNA in spleen lymphocytes was positively correlated with spleen lymphocytes Caspase-3 mRNA and serum Caspase-3 levels(P<0.05).Conclusion Increased expression of PERK is associated with apoptosis of SAP lymphocytes and may be a target for SAP treatment.
引文
[1]Forsmark CE,Vege SS,Wilcox CM.Acute Pancreatitis[J].N Engl J Med,2016,375(20):1972-1981.
[2]石占利,孙静,李志会,等.细胞凋亡在重症急性胰腺炎并发急性肺损伤中的作用机制[J].解放军医学杂志,2017,42(11):974-978.
[3]欧春元,李艳美.大剂量参附注射液对重症急性胰腺炎合并脓毒症休克患者HSP70及IL-10表达的影响[J].右江民族医学院学报,2018,40(1):48-51.
[4]余杨梓,傅强,李静.免疫失衡在重症急性胰腺炎患者继发全身感染中的作用[J].天津医药,2012,40(7):672-674.
[5]Reid DW,Tay AS,Sundaram JR,et al.Complementary Roles of GADD34-and CReP-Containing Eukaryotic Initiation Factor 2αPhosphatases during the Unfolded Protein Response[J].Molecular&Cellular Biology,2016,36(13):1868-1880.
[6]Shen Y,Deng X,Xu N,et al.Relationship between the degree of severe acute pancreatitis and patient immunity[J].Surgery Today,2015,45(8):1009-1017.
[7]Duan L,Yu M A,Chi J,et al.The regulatory role of immunosuppressants on immune abnormalities in acute pancreatitis[J].Biomed Rep,2014,2(2):193-198.
[8]覃月秋,廖品琥,周喜汉,等.重症急性胰腺炎Fas表达、淋巴细胞凋亡及与并发脓毒症的关系[J].世界华人消化杂志,2012,20(32):3107-3113.
[9]Qin Y,Pinhu L,You Y,et al.The role of Fas expression on the occurrence of immunosuppression in severe acute pancreatitis[J].Dig Dis Sci,2013,58(11):3300-3307.
[10]Diao J,Liu R,Rong Y,et al.ATG14promotes membrane tethering and fusion of autophagosomes to endolysosomes[J].Nature,2015,520(7548):563-566.
[11]Bastida-Ruiz D,Aguilar E,Ditisheim A,et al.Endoplasmic reticulum stress responses in placentation-A true balancing act[J].Placenta,2017,57:163-169.
[12]李晓娟,夏榕蔓,楼招欢,等.丹参二萜醌通过PERK-EIF2α通路诱导肺癌PC9细胞凋亡的机制研究[J].中华中医药杂志,2017,32(5):1897-1902.
[13]Hiramatsu N,Messah C,Han J,et al.Translational and posttranslational regulation of XIAP by eIF2αand ATF4promotes ER stress-induced cell death during the unfolded protein response[J].Mol Biol Cell,2014,25(9):1411-1420.
[14]Xia S,Wang J,Kalionis B,et al.Genistein protects against acute pancreatitis via activation of an apoptotic pathway mediated through endoplasmic reticulum stress in rats[J].Biochem Biophys Res Commun,2019,509(2):421-428.
[15]Ma S,Shao L,Liu Y,et al.A study of lymphocyte apoptosis and endoplasmic reticulum stress in the development of sepsis and their association with outcome in septic patients[J].Zhonghua Wei Zhong Bing Ji Jiu Yi Xue,2015,27(2):115-120.
[16]陈伟,马磊,杨立山.甘草次酸对哮喘大鼠气道重塑及肺组织Casepase-3、Bax、Bcl-2表达的影响[J].中药药理与临床,2016,32(04):16-19.
[17]B'chir W,Maurin AC,Carraro V,et al.The eIF2α/ATF4pathway is essential for stress-induced autophagy gene expression[J].Nucleic Acids Research,2013,41(16):7683-7699.
[18]Li Y,Guo Y,Tang J,et al.New insights into the roles of CHOP-induced apoptosis in ER stress[J].Acta Biochim Biophys Sin(Shanghai),2015,47(2):146-147.
[2]石占利,孙静,李志会,等.细胞凋亡在重症急性胰腺炎并发急性肺损伤中的作用机制[J].解放军医学杂志,2017,42(11):974-978.
[3]欧春元,李艳美.大剂量参附注射液对重症急性胰腺炎合并脓毒症休克患者HSP70及IL-10表达的影响[J].右江民族医学院学报,2018,40(1):48-51.
[4]余杨梓,傅强,李静.免疫失衡在重症急性胰腺炎患者继发全身感染中的作用[J].天津医药,2012,40(7):672-674.
[5]Reid DW,Tay AS,Sundaram JR,et al.Complementary Roles of GADD34-and CReP-Containing Eukaryotic Initiation Factor 2αPhosphatases during the Unfolded Protein Response[J].Molecular&Cellular Biology,2016,36(13):1868-1880.
[6]Shen Y,Deng X,Xu N,et al.Relationship between the degree of severe acute pancreatitis and patient immunity[J].Surgery Today,2015,45(8):1009-1017.
[7]Duan L,Yu M A,Chi J,et al.The regulatory role of immunosuppressants on immune abnormalities in acute pancreatitis[J].Biomed Rep,2014,2(2):193-198.
[8]覃月秋,廖品琥,周喜汉,等.重症急性胰腺炎Fas表达、淋巴细胞凋亡及与并发脓毒症的关系[J].世界华人消化杂志,2012,20(32):3107-3113.
[9]Qin Y,Pinhu L,You Y,et al.The role of Fas expression on the occurrence of immunosuppression in severe acute pancreatitis[J].Dig Dis Sci,2013,58(11):3300-3307.
[10]Diao J,Liu R,Rong Y,et al.ATG14promotes membrane tethering and fusion of autophagosomes to endolysosomes[J].Nature,2015,520(7548):563-566.
[11]Bastida-Ruiz D,Aguilar E,Ditisheim A,et al.Endoplasmic reticulum stress responses in placentation-A true balancing act[J].Placenta,2017,57:163-169.
[12]李晓娟,夏榕蔓,楼招欢,等.丹参二萜醌通过PERK-EIF2α通路诱导肺癌PC9细胞凋亡的机制研究[J].中华中医药杂志,2017,32(5):1897-1902.
[13]Hiramatsu N,Messah C,Han J,et al.Translational and posttranslational regulation of XIAP by eIF2αand ATF4promotes ER stress-induced cell death during the unfolded protein response[J].Mol Biol Cell,2014,25(9):1411-1420.
[14]Xia S,Wang J,Kalionis B,et al.Genistein protects against acute pancreatitis via activation of an apoptotic pathway mediated through endoplasmic reticulum stress in rats[J].Biochem Biophys Res Commun,2019,509(2):421-428.
[15]Ma S,Shao L,Liu Y,et al.A study of lymphocyte apoptosis and endoplasmic reticulum stress in the development of sepsis and their association with outcome in septic patients[J].Zhonghua Wei Zhong Bing Ji Jiu Yi Xue,2015,27(2):115-120.
[16]陈伟,马磊,杨立山.甘草次酸对哮喘大鼠气道重塑及肺组织Casepase-3、Bax、Bcl-2表达的影响[J].中药药理与临床,2016,32(04):16-19.
[17]B'chir W,Maurin AC,Carraro V,et al.The eIF2α/ATF4pathway is essential for stress-induced autophagy gene expression[J].Nucleic Acids Research,2013,41(16):7683-7699.
[18]Li Y,Guo Y,Tang J,et al.New insights into the roles of CHOP-induced apoptosis in ER stress[J].Acta Biochim Biophys Sin(Shanghai),2015,47(2):146-147.