中华急诊医学杂志
中華急診醫學雜誌
중화급진의학잡지
CHINESE JOURNAL OF EMERGENCY MEDICINE
2009年
6期
618-622
,共5页
王映珍%王世文%高宁%孙志江%贾晓冬%吕苗
王映珍%王世文%高寧%孫誌江%賈曉鼕%呂苗
왕영진%왕세문%고저%손지강%가효동%려묘
胰岛素样生长因子-1%重症急性胰腺炎%凋亡%Bax%Bcl-2%Caspase-3
胰島素樣生長因子-1%重癥急性胰腺炎%凋亡%Bax%Bcl-2%Caspase-3
이도소양생장인자-1%중증급성이선염%조망%Bax%Bcl-2%Caspase-3
Insulin-like growth factor Ⅰ%Severe acute pancreatitis%Apotosis%Bax%Bcl-2%Caspase-3
目的 研究外源性胰岛素样生长因子-Ⅰ(IGF-Ⅰ)对重症急性胰腺炎(SAP)大鼠肠黏膜上皮细胞凋亡及凋亡相关基因bax,bcl-2,caspase-3 mRNA表达的影响.方法 72只雄性Wistar大鼠按照随机数字表法分为假手术组(SO组)、重症急性胰腺炎组(SAP组)、IGF-Ⅰ治疗组(IGF-Ⅰ组)共三组.通过胰胆管逆行注射5%牛磺胆酸钠制作大鼠SAP模型,SO组用同样方法经胰胆管逆行注射生理盐水;IGF-Ⅰ组分别于术后30 min和术后3 h经后肢内侧皮下注射ICF-Ⅰ.各组动物分别于术后6,12,24 h处死8只,检测血浆淀粉酶,TUNEL法检测小肠黏膜上皮细胞凋亡,观察小肠组织病理学变化并进行病理评分,逆转录聚合酶链反应(RT-PCR)法检测小肠黏膜上皮细胞中bax,bcl-2和caspase-3mRNA的表达.数据采用SPSS 11.0统计软件分析,多组间比较采用单因素方差分析.结果 IGF-Ⅰ治疗组与SAP组各时相点相比,血浆淀粉酶和小肠病理评分均低于各相应时相点,于12 h和24 h差异有统计学意义,均P<0.05;小肠黏膜上皮细胞凋亡指数IGF-Ⅰ组与SAP组各相应时相点相比均显著降低[6 h:(13.88±1.73)vs.(19.00±2.78);12 h:(10.13±1.55)vs.(17.63±1.60);24 h:(9.50±1.07)vs.(17.25±2.76)],P<0.05;电镜示小肠组织病理变化较SAP组明显改善.bax mRNA表达在SAP组各时点较SO组相应时相点明显增高[6 h:(1.35±0.18)vs.(0.85±0.12);12 h:(1.21±0.21)vs.(0.86±0.24);24 h:(1.14±0.24)vs.(0.95±0.22)],6 h即达高峰,而在IGF-Ⅰ组的表达与SAP组相应时相点比较则明显减弱,在12 h和24 h差异具有统计学意义(P<0.05),于24 h即接近SO组;caspase-3 mRNA表达在SAP组各时相点较SO组相应时相点明显增高[6 h:(0.78±0.01)vs.(0.55±0.04);12 h:(0.79±0.04)vs.(0.57±0.05);24 h:(0.81±0.06)vs.(0.55±0.01)(P<0.01)],而在IGF-Ⅰ组的表达与SAP组相应时相点比较则明显减弱,在12 h和24 h差异具有统计学意义(P<0.05):bcl-2 mRNA的表达在SO组和SAP组均较弱且差异无统计学意义,P>0.05,而在IGF-Ⅰ组各时相点的表达则明显增强[6 h:(0.65±0.07)vs.(0.54±0.04)vs.(0.57±0.06);12 h:(0.69±0.04)vs.(0.56±0.05)vs.(0.53±0.05);24 h:(0.72±0.05)vs.(0.54±0.07)vs.(0.58±0.08),P<0.05].结论 外源性IGF-Ⅰ通过拮抗SAP大鼠小肠黏膜上皮细胞的凋亡从而可以减轻SAP大鼠肠黏膜损害,其机制可能与其上调bcl-2 mRNA表达及下调bax,caspase-3 mRNA表达有关.
目的 研究外源性胰島素樣生長因子-Ⅰ(IGF-Ⅰ)對重癥急性胰腺炎(SAP)大鼠腸黏膜上皮細胞凋亡及凋亡相關基因bax,bcl-2,caspase-3 mRNA錶達的影響.方法 72隻雄性Wistar大鼠按照隨機數字錶法分為假手術組(SO組)、重癥急性胰腺炎組(SAP組)、IGF-Ⅰ治療組(IGF-Ⅰ組)共三組.通過胰膽管逆行註射5%牛磺膽痠鈉製作大鼠SAP模型,SO組用同樣方法經胰膽管逆行註射生理鹽水;IGF-Ⅰ組分彆于術後30 min和術後3 h經後肢內側皮下註射ICF-Ⅰ.各組動物分彆于術後6,12,24 h處死8隻,檢測血漿澱粉酶,TUNEL法檢測小腸黏膜上皮細胞凋亡,觀察小腸組織病理學變化併進行病理評分,逆轉錄聚閤酶鏈反應(RT-PCR)法檢測小腸黏膜上皮細胞中bax,bcl-2和caspase-3mRNA的錶達.數據採用SPSS 11.0統計軟件分析,多組間比較採用單因素方差分析.結果 IGF-Ⅰ治療組與SAP組各時相點相比,血漿澱粉酶和小腸病理評分均低于各相應時相點,于12 h和24 h差異有統計學意義,均P<0.05;小腸黏膜上皮細胞凋亡指數IGF-Ⅰ組與SAP組各相應時相點相比均顯著降低[6 h:(13.88±1.73)vs.(19.00±2.78);12 h:(10.13±1.55)vs.(17.63±1.60);24 h:(9.50±1.07)vs.(17.25±2.76)],P<0.05;電鏡示小腸組織病理變化較SAP組明顯改善.bax mRNA錶達在SAP組各時點較SO組相應時相點明顯增高[6 h:(1.35±0.18)vs.(0.85±0.12);12 h:(1.21±0.21)vs.(0.86±0.24);24 h:(1.14±0.24)vs.(0.95±0.22)],6 h即達高峰,而在IGF-Ⅰ組的錶達與SAP組相應時相點比較則明顯減弱,在12 h和24 h差異具有統計學意義(P<0.05),于24 h即接近SO組;caspase-3 mRNA錶達在SAP組各時相點較SO組相應時相點明顯增高[6 h:(0.78±0.01)vs.(0.55±0.04);12 h:(0.79±0.04)vs.(0.57±0.05);24 h:(0.81±0.06)vs.(0.55±0.01)(P<0.01)],而在IGF-Ⅰ組的錶達與SAP組相應時相點比較則明顯減弱,在12 h和24 h差異具有統計學意義(P<0.05):bcl-2 mRNA的錶達在SO組和SAP組均較弱且差異無統計學意義,P>0.05,而在IGF-Ⅰ組各時相點的錶達則明顯增彊[6 h:(0.65±0.07)vs.(0.54±0.04)vs.(0.57±0.06);12 h:(0.69±0.04)vs.(0.56±0.05)vs.(0.53±0.05);24 h:(0.72±0.05)vs.(0.54±0.07)vs.(0.58±0.08),P<0.05].結論 外源性IGF-Ⅰ通過拮抗SAP大鼠小腸黏膜上皮細胞的凋亡從而可以減輕SAP大鼠腸黏膜損害,其機製可能與其上調bcl-2 mRNA錶達及下調bax,caspase-3 mRNA錶達有關.
목적 연구외원성이도소양생장인자-Ⅰ(IGF-Ⅰ)대중증급성이선염(SAP)대서장점막상피세포조망급조망상관기인bax,bcl-2,caspase-3 mRNA표체적영향.방법 72지웅성Wistar대서안조수궤수자표법분위가수술조(SO조)、중증급성이선염조(SAP조)、IGF-Ⅰ치료조(IGF-Ⅰ조)공삼조.통과이담관역행주사5%우광담산납제작대서SAP모형,SO조용동양방법경이담관역행주사생리염수;IGF-Ⅰ조분별우술후30 min화술후3 h경후지내측피하주사ICF-Ⅰ.각조동물분별우술후6,12,24 h처사8지,검측혈장정분매,TUNEL법검측소장점막상피세포조망,관찰소장조직병이학변화병진행병리평분,역전록취합매련반응(RT-PCR)법검측소장점막상피세포중bax,bcl-2화caspase-3mRNA적표체.수거채용SPSS 11.0통계연건분석,다조간비교채용단인소방차분석.결과 IGF-Ⅰ치료조여SAP조각시상점상비,혈장정분매화소장병리평분균저우각상응시상점,우12 h화24 h차이유통계학의의,균P<0.05;소장점막상피세포조망지수IGF-Ⅰ조여SAP조각상응시상점상비균현저강저[6 h:(13.88±1.73)vs.(19.00±2.78);12 h:(10.13±1.55)vs.(17.63±1.60);24 h:(9.50±1.07)vs.(17.25±2.76)],P<0.05;전경시소장조직병리변화교SAP조명현개선.bax mRNA표체재SAP조각시점교SO조상응시상점명현증고[6 h:(1.35±0.18)vs.(0.85±0.12);12 h:(1.21±0.21)vs.(0.86±0.24);24 h:(1.14±0.24)vs.(0.95±0.22)],6 h즉체고봉,이재IGF-Ⅰ조적표체여SAP조상응시상점비교칙명현감약,재12 h화24 h차이구유통계학의의(P<0.05),우24 h즉접근SO조;caspase-3 mRNA표체재SAP조각시상점교SO조상응시상점명현증고[6 h:(0.78±0.01)vs.(0.55±0.04);12 h:(0.79±0.04)vs.(0.57±0.05);24 h:(0.81±0.06)vs.(0.55±0.01)(P<0.01)],이재IGF-Ⅰ조적표체여SAP조상응시상점비교칙명현감약,재12 h화24 h차이구유통계학의의(P<0.05):bcl-2 mRNA적표체재SO조화SAP조균교약차차이무통계학의의,P>0.05,이재IGF-Ⅰ조각시상점적표체칙명현증강[6 h:(0.65±0.07)vs.(0.54±0.04)vs.(0.57±0.06);12 h:(0.69±0.04)vs.(0.56±0.05)vs.(0.53±0.05);24 h:(0.72±0.05)vs.(0.54±0.07)vs.(0.58±0.08),P<0.05].결론 외원성IGF-Ⅰ통과길항SAP대서소장점막상피세포적조망종이가이감경SAP대서장점막손해,기궤제가능여기상조bcl-2 mRNA표체급하조bax,caspase-3 mRNA표체유관.
Objective To investigate the effects of exogenous IGF- Ⅰon apoptosis, bax,bcl-2 and caspase-3 gene mRNA transcription in intestinal mucosal epithelial cell of SAP rats. Method Seventy-two male Wistar rats were randomly divided into sham operation group (SO),SAP group(SAP) and IGF-Ⅰ treatment (SAP + IGF-Ⅰ) group.Every group was randomly divided into 3 time units (6,12,24 h),8 rats as each time unit. SAP was induced in the rats by injecting adversely 5.0 % sodium taurocholate into biliary-pancreafic duct. The SO rats were infused with NS by the same way. The rats in IGF-Ⅰgroup were injected with IGF-Ⅰ by subcutano at half an hour before operation and three hours after operation,respectively. Animals in each group were killed separately at 6,12 and 24 hours after operation.The apoptosis in mucesal cells of small intestine was detected by TUNEL, and histo pathological changes of the small intestine was observed. The expressions of bax and bcl-2 and caspase-3 mR-NA gene in small intestine were measured by reverse transcription polymerase chain reaction(RT-PCR). Results Compared with the SAP group,the serum amylase were lower in IGF-Ⅰ group,and there existed significant at 12 h and24 h (P < 0.05).The pathological score of small intestinal was significantly reduced in IGF-Ⅰ group com-pared with SAP group,and there were statistical differences at 12 h and 24 h.ln IGFo-Ⅰ group,the apoptosis index of intestinal epithelial decreased significantly compared with SAP group[6 h: (13.88±1.73) vs. (19.00±2.78) ;12h:(10.13±1.55) vs. (17.63±.60);24 h:(9.50±1.07) vs. (17.25±2.76)] (P <0.05); the histopathdogical changes were more improved compared wit SAP group under the electronic microscope; the expres-sion of bax mRNA [6 h:(1.35±0.18) vs.(0.85±0.12);12 h:(1.21±0.21) vs. (0.86±0.24);24 h:(1.14±0.24) vs. (0.95±0.22)] and caspese-3 mRNA[6 h:(0.78±0.01) vs. (0.55±0.04);12 h:(0.79±0.04) vs. (0.57±0.05) ;24 h: (0.81±0.06) vs. (0.55±0.01) (P < 0.01)] were higher in three time units in SAP group than those in SO group (P < 0.01) ,and in IGF-1 group it was weakened significantly compared with the SAP group at each time point (P <0.05). bcl-2 mRNA expression was weak and have no difference between the SO group and SAP group (P > 0.05), but increased signifycantly in the IGF-± group at each time point [6 h:(0.65±0.07) vs. (0.54±0.04) vs. (0.57±0.06);12 h:(0.69±0.04) vs. (0.56±0.05) vs. (0.53±0.05);24h:(0.72±0.05) vs. (0.54±0.07) vs. (0.58±0.08)] (P <0.05). Conclusions Exogenous IGF-Ⅰ could rivalry SAP induced apoptosis to mucosal cells of small intestine , then could alleviate SAP induced injury to intestinal mucosal, It may be associated with the mechanisms that IGF-Ⅰ could improve the expression of bcl-2 mRNA and inhibit the expression of bax,caspase-3 mRNA.
