中华胰腺病杂志
中華胰腺病雜誌
중화이선병잡지
CHINESE JOURNAL OF PANCREATOLOGY
2011年
6期
417-419
,共3页
胰腺炎%磷脂酰肌醇3-激酶γ%蛙皮素%自噬
胰腺炎%燐脂酰肌醇3-激酶γ%蛙皮素%自噬
이선염%린지선기순3-격매γ%와피소%자서
Pancreatitis%Phosphoinositide 3-Kinase-C2-gamma%Cerulein%Autophagy
目的 探讨磷脂酰肌醇3-激酶γ(PI3Kγ)对小鼠实验性急性胰腺炎胰腺腺泡细胞自噬作用的影响,并探讨其意义.方法 野生型C57 BL/6小鼠和PI3Kγ基因敲除小鼠各18只,按数字表法随机分为对照组(6只)和急性胰腺炎(AP)组(12只),采用蛙皮素50μg/kg体重腹腔内注射7次、每次间隔1h的方法制备AP模型.首次注射后7h处死小鼠,光镜下观察胰腺病理学变化,免疫荧光检测自噬泡主要组成蛋白LC3颗粒,荧光分光光度计测定胰蛋白酶活性,蛋白质印迹法检测自噬相关蛋白beclin1、LC3-Ⅱ和p62的表达.结果 AP组野生型小鼠和PI3Kγ基因敲除小鼠的胰腺自噬空泡数量分别为(5.14±0.85)、(2.25±0.54)个/每高倍视野(HPF),LC3荧光免疫颗粒数量分别为(78.6±9.38)、(26.4±4.21)个/HPF,胰蛋白酶活性分别为(0.827±0.126)、(0.358±0.098) pmol/mg蛋白,各组间差异均具有统计学意义(P值均＜0.05).野生型小鼠的p62蛋白表达较PI3Kγ基因敲除小鼠显著减弱(0.11比0.92,P＜0.05),面LC3 -Ⅱ、beclin1蛋白表达较PI3Kγ基因敲除小鼠显著增强(1.82比0.93,1.43比1.05,P值均＜0.05).结论 AP时PI3Kγ可能通过增强小鼠胰腺腺泡细胞的自噬作用,促进胰蛋白酶原的活化及诱导腺泡细胞坏死.
目的 探討燐脂酰肌醇3-激酶γ(PI3Kγ)對小鼠實驗性急性胰腺炎胰腺腺泡細胞自噬作用的影響,併探討其意義.方法 野生型C57 BL/6小鼠和PI3Kγ基因敲除小鼠各18隻,按數字錶法隨機分為對照組(6隻)和急性胰腺炎(AP)組(12隻),採用蛙皮素50μg/kg體重腹腔內註射7次、每次間隔1h的方法製備AP模型.首次註射後7h處死小鼠,光鏡下觀察胰腺病理學變化,免疫熒光檢測自噬泡主要組成蛋白LC3顆粒,熒光分光光度計測定胰蛋白酶活性,蛋白質印跡法檢測自噬相關蛋白beclin1、LC3-Ⅱ和p62的錶達.結果 AP組野生型小鼠和PI3Kγ基因敲除小鼠的胰腺自噬空泡數量分彆為(5.14±0.85)、(2.25±0.54)箇/每高倍視野(HPF),LC3熒光免疫顆粒數量分彆為(78.6±9.38)、(26.4±4.21)箇/HPF,胰蛋白酶活性分彆為(0.827±0.126)、(0.358±0.098) pmol/mg蛋白,各組間差異均具有統計學意義(P值均＜0.05).野生型小鼠的p62蛋白錶達較PI3Kγ基因敲除小鼠顯著減弱(0.11比0.92,P＜0.05),麵LC3 -Ⅱ、beclin1蛋白錶達較PI3Kγ基因敲除小鼠顯著增彊(1.82比0.93,1.43比1.05,P值均＜0.05).結論 AP時PI3Kγ可能通過增彊小鼠胰腺腺泡細胞的自噬作用,促進胰蛋白酶原的活化及誘導腺泡細胞壞死.
목적 탐토린지선기순3-격매γ(PI3Kγ)대소서실험성급성이선염이선선포세포자서작용적영향,병탐토기의의.방법 야생형C57 BL/6소서화PI3Kγ기인고제소서각18지,안수자표법수궤분위대조조(6지)화급성이선염(AP)조(12지),채용와피소50μg/kg체중복강내주사7차、매차간격1h적방법제비AP모형.수차주사후7h처사소서,광경하관찰이선병이학변화,면역형광검측자서포주요조성단백LC3과립,형광분광광도계측정이단백매활성,단백질인적법검측자서상관단백beclin1、LC3-Ⅱ화p62적표체.결과 AP조야생형소서화PI3Kγ기인고제소서적이선자서공포수량분별위(5.14±0.85)、(2.25±0.54)개/매고배시야(HPF),LC3형광면역과립수량분별위(78.6±9.38)、(26.4±4.21)개/HPF,이단백매활성분별위(0.827±0.126)、(0.358±0.098) pmol/mg단백,각조간차이균구유통계학의의(P치균＜0.05).야생형소서적p62단백표체교PI3Kγ기인고제소서현저감약(0.11비0.92,P＜0.05),면LC3 -Ⅱ、beclin1단백표체교PI3Kγ기인고제소서현저증강(1.82비0.93,1.43비1.05,P치균＜0.05).결론 AP시PI3Kγ가능통과증강소서이선선포세포적자서작용,촉진이단백매원적활화급유도선포세포배사.
Objective To investigate the influence of phosphoinositide 3-Kinase-C2-gamma (PI3Kγ)on pancreas acinar cells autophagy in experimental acute pancreatitis in mice and explore its significance.Methods Eighteen C57BL/6 wild type (WT) and eighteen PI3Kγ knockout (KO) mice were randomly divided into control group (n =6) and acute panereatitis (AP) group (n =12),respectively.AP models were induced by intraperitoneal injection of 50 μg cerulein/kg body weight,once the other hour for seven times.The mice were sacrificed 7 hours after model induction.The pathological changes of the pancreas were observed through microscope,LC3 dots were determined by immunofluorescence,the trypsin activity was measured by fluorescence spectrophotometer,and the expression of autophagy related protein beclin1,p62 and LC3- Ⅱ were measured by Western blot.Results The autophagy vacuoles counts in pancreatic tissue of WT mice and KO mice were (5.14 ±0.85),(2.25 ±0.54)/HPF,the LC3 immunofluorescence dots counts were (78.6 ±9.38),( 26.4 ± 4.21 )/HPF,the trypsin activities were ( 0.827 ± 0.126 ),( 0.358 ± 0.098 ) pmol/mg protein,the difference between the two groups was statistically significant ( P ＜ 0.05 ).The p62 protein expression was greatly decreased in WT mice compared with their KO counterpart (0.11 vs 0.92,P ＜ 0.05 ),while the expressions of LC3 Ⅱ,beclin1 were greatly increased in WT mice compared with their KO counterpart ( 1.82 vs 0.93,1.43 vs 1.05,P ＜ 0.05 ).Conclusions PI 3 Kγmay up- regulate autophagy of pancreatic acinar cells during acute pancreatitis in mice,then promote trypsinogen activation and necrosis of acinar cells.
