中华急诊医学杂志
中華急診醫學雜誌
중화급진의학잡지
CHINESE JOURNAL OF EMERGENCY MEDICINE
2014年
4期
389-392
,共4页
小鼠%急性酒精性肝损伤%凋亡%增殖%T-ERK%P-ERK%蛋白激酶C%P-PKC%caspase-3
小鼠%急性酒精性肝損傷%凋亡%增殖%T-ERK%P-ERK%蛋白激酶C%P-PKC%caspase-3
소서%급성주정성간손상%조망%증식%T-ERK%P-ERK%단백격매C%P-PKC%caspase-3
Mice%Acute alcoholic liver injury%Apoptosis%Proliferation%T-ERK%P-ERK%PKC%P-PKC%caspase-3
目的 建立小鼠急性酒精性肝损伤的模型,通过测定肝脏组织内的相关增殖凋亡蛋白的变化,探讨抑制其细胞凋亡的治疗时间窗,以指导临床治疗肝损伤.方法 将30只雄性KM种小鼠饲养于首都医科大学清洁级动物房,随机分为两组(随机数字法).对照组10只给予正常喂养,实验组20只,一次性给予50％乙醇(12 mL/kg)灌胃,分别在灌胃6h (m6h)和12 h(m12 h)两个时间点处死小鼠各10只.苏木素-伊红(HE)染色法观察小鼠肝脏组织病理学的变化.应用Western-blot的方法监测小鼠肝脏中蛋白T-ERK,P-ERK,PKC,P-PKC,caspase-3的含量变化.用SPSS 11.5统计软件对数据进行处理,进行方差分析,以P＜0.05为差异具有统计学意义.结果 通过观察小鼠灌酒后的行为变化,结合形态学指标验证小鼠急性酒精性肝损伤的模型复制成功,小鼠无死亡,实验组小鼠出现嗜睡、行动迟缓等醉酒状态.实验组经酒精灌胃6h所测指标较对照组没有统计学意义,而经酒精灌胃12 h,肝小叶结构紊乱,肝细胞内脂肪空泡形成,与对照组比较,P-ERK和P-PKC显著降低[(2.41±0.38), (0.97±0.25),F=4.82,P＜0.05;(0.16±0.00),(0.08±0.01),F=29.63,P＜0.05],caspase-3显著升高[(0.30±0.02),(0.11±0.01),F=34.38,P＜0.05].结论 给予小鼠大剂量酒精灌胃后,凋亡主要集中在12 h出现,因此抑制小鼠急性酒精性肝损伤细胞凋亡的时间窗可能存在于6～12h之间.
目的 建立小鼠急性酒精性肝損傷的模型,通過測定肝髒組織內的相關增殖凋亡蛋白的變化,探討抑製其細胞凋亡的治療時間窗,以指導臨床治療肝損傷.方法 將30隻雄性KM種小鼠飼養于首都醫科大學清潔級動物房,隨機分為兩組(隨機數字法).對照組10隻給予正常餵養,實驗組20隻,一次性給予50％乙醇(12 mL/kg)灌胃,分彆在灌胃6h (m6h)和12 h(m12 h)兩箇時間點處死小鼠各10隻.囌木素-伊紅(HE)染色法觀察小鼠肝髒組織病理學的變化.應用Western-blot的方法鑑測小鼠肝髒中蛋白T-ERK,P-ERK,PKC,P-PKC,caspase-3的含量變化.用SPSS 11.5統計軟件對數據進行處理,進行方差分析,以P＜0.05為差異具有統計學意義.結果 通過觀察小鼠灌酒後的行為變化,結閤形態學指標驗證小鼠急性酒精性肝損傷的模型複製成功,小鼠無死亡,實驗組小鼠齣現嗜睡、行動遲緩等醉酒狀態.實驗組經酒精灌胃6h所測指標較對照組沒有統計學意義,而經酒精灌胃12 h,肝小葉結構紊亂,肝細胞內脂肪空泡形成,與對照組比較,P-ERK和P-PKC顯著降低[(2.41±0.38), (0.97±0.25),F=4.82,P＜0.05;(0.16±0.00),(0.08±0.01),F=29.63,P＜0.05],caspase-3顯著升高[(0.30±0.02),(0.11±0.01),F=34.38,P＜0.05].結論 給予小鼠大劑量酒精灌胃後,凋亡主要集中在12 h齣現,因此抑製小鼠急性酒精性肝損傷細胞凋亡的時間窗可能存在于6～12h之間.
목적 건립소서급성주정성간손상적모형,통과측정간장조직내적상관증식조망단백적변화,탐토억제기세포조망적치료시간창,이지도림상치료간손상.방법 장30지웅성KM충소서사양우수도의과대학청길급동물방,수궤분위량조(수궤수자법).대조조10지급여정상위양,실험조20지,일차성급여50％을순(12 mL/kg)관위,분별재관위6h (m6h)화12 h(m12 h)량개시간점처사소서각10지.소목소-이홍(HE)염색법관찰소서간장조직병이학적변화.응용Western-blot적방법감측소서간장중단백T-ERK,P-ERK,PKC,P-PKC,caspase-3적함량변화.용SPSS 11.5통계연건대수거진행처리,진행방차분석,이P＜0.05위차이구유통계학의의.결과 통과관찰소서관주후적행위변화,결합형태학지표험증소서급성주정성간손상적모형복제성공,소서무사망,실험조소서출현기수、행동지완등취주상태.실험조경주정관위6h소측지표교대조조몰유통계학의의,이경주정관위12 h,간소협결구문란,간세포내지방공포형성,여대조조비교,P-ERK화P-PKC현저강저[(2.41±0.38), (0.97±0.25),F=4.82,P＜0.05;(0.16±0.00),(0.08±0.01),F=29.63,P＜0.05],caspase-3현저승고[(0.30±0.02),(0.11±0.01),F=34.38,P＜0.05].결론 급여소서대제량주정관위후,조망주요집중재12 h출현,인차억제소서급성주정성간손상세포조망적시간창가능존재우6～12h지간.
Objective To study the proliferation and apoptosis of related proteins in pathological liver tissues of alcohol-induced mice,and establish a model and time-evolution rule of liver cell apoptosis,which can be used to guide the clinical treatment of acute alcoholic liver injury.Methods A total of 30 male KM mice were fed in a clean grade animal room at the Capital Medical University and then randomly (random number) separated into two groups.The 10 mice in the normal group were fed without ethanol,while the other 20 mice in the experimental group were given a one-time grant of 50％ ethanol (12 mL/kg) by gavage.The mice in the experimental group were killed at two time points,6 h for 10 mice and 12 h for the other 10,after the intragastric administration.Hematoxylin and eosin (HE) staining was used to observe the morphological changes of liver in mice.The concentrations of T-ERK,p-ERK,PKC,p-PKC and caspase-3 were determined by the Western-blot method.The data were analyzed by Analysis of variance (ANOVA) method using statistical software SPSS 11.5 and criterion P ＜ 0.05 is chosen to determine differences that are statistically significant.Results By observing the behavioral changes and morphological indexes of mice,we confirmed the success of the model for acute alcoholic liver injury.During the process of re-building the model,no mice died.The mice in the experimental group appeared in drunken states,such as sleepiness and slowness of movement.Compared to the normal group,the experimental subgroup at the 6 h point showed no difference statistical significant; while the experimental subgroup at the 12 h point showed obvious histological changes in tissues,including the disorder of hepatic lobule structure and fatty vacuolization of hepatocytes.At the same time,in the experimental subgroup at the 12 h point,both P-ERK and P-PKC significantly decreased [(2.41 ±0.38),(0.97 ±0.25),F=4.82,P＜0.05; (0.16 ±0.00),(0.08 ± 0.01),F =29.63,P ＜ 0.05],but caspase-3 significantly increased [(0.30 ± 0.02),(0.11 ± 0.01),F =34.38,P ＜ 0.05].Conclusions In mice after intragastric administration of large doses of alcohol,the hepatic cell apoptosis appeared mainly after 6 h but before 12 h,therefore 6 ～ 12 h might be the time window to inhibit the cell apoptosis of mice' s acute liver injury from alcohol induction.
