CAJ | 학술논문

目的探讨褪黑素(MT)对急性坏死性胰腺炎(ANP)胰腺腺泡细胞钙超载的调节作用及其机制.方法 54只SD大鼠均分为假手术组(SO组)、急性坏死性胰腺炎组(ANP组,胰胆管逆行注射牛磺胆酸钠法造模)和MT组(腹腔注射MT后30 min ANP造模).术后1、4、8h分批处死大鼠并进行胰腺病理检查.用荧光控测法测定胰腺组织内游离钙离子浓度,实时荧光定量PCR及免疫印迹法分别检测胰腺细胞中钙调素依赖性蛋白激酶Ⅱ( CaMKⅡ)mRNA和蛋白表达水平.结果 ANP组随时间延长胰腺病理损伤进行性加重,MT组胰腺病理损伤较ANP组明显减轻(1、4、8h时病理评分比较的t值分别为-7.95、-9.72、-7.69,P值均=0.00).同时间点比较,ANP组胰腺组织内游离钙离子浓度均较SO组明显增高(1、4、8h时的t值分别=13.09、18.58、16.56,P值均=0.00),MT组较SO组略高,但较ANP组明显降低(1、4、8h时的τ值分别=-10.03、-11.75、-11.02,P值均=0.00).与SO组相比,ANP组CaMKⅡmRNA及蛋白表达水平均明显升高,MT对其表达有明显的抑制作用.结论 MT干预可抑制CaMKⅡ表达,从而减轻胰腺腺泡内钙超载,起到保护胰腺组织的作用.
목적 탐토퇴흑소(MT)대급성배사성이선염(ANP)이선선포세포개초재적조절작용급기궤제.방법 54지SD대서균분위가수술조(SO조)、급성배사성이선염조(ANP조,이담관역행주사우광담산납법조모)화MT조(복강주사MT후30 min ANP조모).술후1、4、8h분비처사대서병진행이선병리검사.용형광공측법측정이선조직내유리개리자농도,실시형광정량PCR급면역인적법분별검측이선세포중개조소의뢰성단백격매Ⅱ( CaMKⅡ)mRNA화단백표체수평.결과 ANP조수시간연장이선병리손상진행성가중,MT조이선병리손상교ANP조명현감경(1、4、8h시병리평분비교적t치분별위-7.95、-9.72、-7.69,P치균=0.00).동시간점비교,ANP조이선조직내유리개리자농도균교SO조명현증고(1、4、8h시적t치분별=13.09、18.58、16.56,P치균=0.00),MT조교SO조략고,단교ANP조명현강저(1、4、8h시적τ치분별=-10.03、-11.75、-11.02,P치균=0.00).여SO조상비,ANP조CaMKⅡmRNA급단백표체수평균명현승고,MT대기표체유명현적억제작용.결론 MT간예가억제CaMKⅡ표체,종이감경이선선포내개초재,기도보호이선조직적작용.
Objective To investigate the regulation and the mechanism of Melatonin in calcium overload of pancreatic acinar cell in acute necrotizing pancreatitis (ANP). Methods Fifty-four Sprague-Dawley (SD) rats were equally divided into three groups:sham-operation group (SO group),ANP group (created with retrograde cholangiopancreatography injection of sodium taurocholate) and MT group (ANP model made after intra-peritoneal injection MT for 30 mins).Rats were sacrificed at 1,4 and 8hours after operation and pancreas tissues were underwent pathological examination.The free calcium concentration of pancreas tissues was determined by fluorescence minitoring method; and the expression of CaMK Ⅱ in pancreas tissues at mRNA and protein level was tested by real-time PCR and Western Blot.Results Pancreatic pathological injury in ANP groups was progressively severe as time extended,which was obviously ameliorated in MT group compared with ANP group (the t value of compared pathological score at 1,4 and 8 hour was:-7.95,-9.72 and -7.69,all P=0.00).Compared at same time point,the free Ca2+ concentration of pancreas tissues in ANP group was significantly higher than that of SO group (the t value of 1,4 and 8 hour was 13.09,18.58 and 16.56,all P=0.00).It was a little bit higher in MT group compared with that of SO group,however was significantly lower than that of ANP group (the t value of 1,4 and 8 hour was -10.03,-11.75 and -11.02,all P =0.00).Compared with SO group,the expression of CaMK Ⅱ at mRNA and protein level significantly increased in ANP group; MT significantly inhibited its expression.Conclusions The expression of CaMK Ⅱ may be inhibited by MT interfere,and then lower the calcium overload in pancreatic acinar cell,which play a role in pancreas tissue protection.