中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2013年
1期
29-32
,共4页
坏死性胰腺炎%脱噬作用%蛋白组学
壞死性胰腺炎%脫噬作用%蛋白組學
배사성이선염%탈서작용%단백조학
Necrotizing pancreatitis%Apoptosis%Proteomics
目的 观察急性坏死性胰腺炎(ANP)大鼠外周血中性粒细胞(PMN)蛋白组学的变化.方法 SD大鼠60只,随机分为2组,每组30只,建立ANP组和SO(假手术)组模型,分别在制模后3、6、12 h分批处死大鼠,抽取下腔静脉血测定血淀粉酶含量,密度梯度离心分离PMN,用流式细胞仪测定PMN凋亡比率.切取胰腺标本组织,病理切片苏木素-伊红(HE)染色,并对胰腺组织进行病理评分.同时对大鼠制模后12h的PMN裂解,应用Label free技术对两组PMN进行蛋白定量差异分析,DeCyder MSTM 2.0分析定量数据,MASCOT算法鉴定定量多肽分子,且进一步鉴定出与凋亡有关的差异蛋白质.结果 ANP组PMN凋亡延迟,至12h达到最低(2.15 ±0.45),与SO组各个时间点比较差异有统计学意义(P＜0.01)；鉴定出与PMN凋亡有关的4种蛋白质相对分子质量为78000葡萄糖调节蛋白、RhoGTPases、L-乳酸脱氢酶A链和血红蛋白α2链(ANP组与SO组的比值分别为1.953614、3.526625、1.766764、0.609825),两组差异有统计学意义(P＜0.05).结论 ANP时,PMN凋亡延迟,与凋亡有关的差异蛋白质可能参与了ANP外周血PMN凋亡延迟.
目的 觀察急性壞死性胰腺炎(ANP)大鼠外週血中性粒細胞(PMN)蛋白組學的變化.方法 SD大鼠60隻,隨機分為2組,每組30隻,建立ANP組和SO(假手術)組模型,分彆在製模後3、6、12 h分批處死大鼠,抽取下腔靜脈血測定血澱粉酶含量,密度梯度離心分離PMN,用流式細胞儀測定PMN凋亡比率.切取胰腺標本組織,病理切片囌木素-伊紅(HE)染色,併對胰腺組織進行病理評分.同時對大鼠製模後12h的PMN裂解,應用Label free技術對兩組PMN進行蛋白定量差異分析,DeCyder MSTM 2.0分析定量數據,MASCOT算法鑒定定量多肽分子,且進一步鑒定齣與凋亡有關的差異蛋白質.結果 ANP組PMN凋亡延遲,至12h達到最低(2.15 ±0.45),與SO組各箇時間點比較差異有統計學意義(P＜0.01)；鑒定齣與PMN凋亡有關的4種蛋白質相對分子質量為78000葡萄糖調節蛋白、RhoGTPases、L-乳痠脫氫酶A鏈和血紅蛋白α2鏈(ANP組與SO組的比值分彆為1.953614、3.526625、1.766764、0.609825),兩組差異有統計學意義(P＜0.05).結論 ANP時,PMN凋亡延遲,與凋亡有關的差異蛋白質可能參與瞭ANP外週血PMN凋亡延遲.
목적 관찰급성배사성이선염(ANP)대서외주혈중성립세포(PMN)단백조학적변화.방법 SD대서60지,수궤분위2조,매조30지,건립ANP조화SO(가수술)조모형,분별재제모후3、6、12 h분비처사대서,추취하강정맥혈측정혈정분매함량,밀도제도리심분리PMN,용류식세포의측정PMN조망비솔.절취이선표본조직,병리절편소목소-이홍(HE)염색,병대이선조직진행병리평분.동시대대서제모후12h적PMN렬해,응용Label free기술대량조PMN진행단백정량차이분석,DeCyder MSTM 2.0분석정량수거,MASCOT산법감정정량다태분자,차진일보감정출여조망유관적차이단백질.결과 ANP조PMN조망연지,지12h체도최저(2.15 ±0.45),여SO조각개시간점비교차이유통계학의의(P＜0.01)；감정출여PMN조망유관적4충단백질상대분자질량위78000포도당조절단백、RhoGTPases、L-유산탈경매A련화혈홍단백α2련(ANP조여SO조적비치분별위1.953614、3.526625、1.766764、0.609825),량조차이유통계학의의(P＜0.05).결론 ANP시,PMN조망연지,여조망유관적차이단백질가능삼여료ANP외주혈PMN조망연지.
Objective To observe the proteomic changes of peripheral blood neutrophils (PMN)in rats with acute necrotizing pancreatitis (ANP).Methods Sixty SD rats were randomly divided into two groups (n =30 each):ANP group and sham-operated (SO) group.The rats were sacrificed at 3rd,6th,and 12 h.The venous blood serum was taken from the inferior vena for determination of amylase content.Density gradient centrifugation was used to isolatie PMN.Apoptosis ratio of PMN was detected by using flow cytometry.The pathological sections from the excised pancreas tissue samples were subjected to hematoxylin and eosin (HE) staining and pathologically scored.PMN lysis molding in rats after 12 h at the same time,the application of Label free two groups of PMN the protein quantitative differences analysis using DeCyder MSTM 2.0 analyze quantitative data,identification with MASCOT algorithm quantitative polypeptide molecules,and further identified with apoptosis related differences in protein.Results ANP group PMN apoptosis delay,the lowest (2.15 ±0.45) to 12 h,the difference was statistically significant (P ＜ 0.01) compared with the SO group at each time point;identified four PMN apoptosis protein:78 000glucose regulatory proteins,the ratio of the the RhoGTPases,L-lactate dehydrogenase A chain and hemoglobin α2 chain (ANP group and SO group,respectively 1.953 614,3.526 625,1.766 764,0.609 825),the difference was statistically significant (P ＜ 0.05).Conclusion ANP,PMN apoptosis delay apoptosis-related differences in the protein may be involved in the ANP peripheral blood PMN apoptosis delay.
