中华肝胆外科杂志
中華肝膽外科雜誌
중화간담외과잡지
CHINESE JOURNAL OF HEPATOBILIARY SURGERY
2013年
1期
58-61
,共4页
靳杯%刘立民%闫文貌%程石
靳杯%劉立民%閆文貌%程石
근배%류립민%염문모%정석
胰腺炎,急性坏死性%肿瘤抑制蛋白质类%肺%创伤和损伤
胰腺炎,急性壞死性%腫瘤抑製蛋白質類%肺%創傷和損傷
이선염,급성배사성%종류억제단백질류%폐%창상화손상
Pancreatitis,acute necrotizing%Tumor suppressor proteins%Lung%Wounds and injuries
目的 研究肿瘤抑制蛋白CYLD在急性坏死性胰腺炎(ANP)肺损伤中的表达状况及其对核转录因子-κB(NF-κB)的影响,探讨其在肺损伤中的作用.方法 72只成年SD大鼠随机分成假手术组、ANP组及氯化钆(GdC13)治疗组,每组24只.采用逆行胰胆管注射5％牛磺酸钠的方法建立ANP大鼠模型.各组术后1、3、6、12 h分批处死动物.经支气管肺泡灌洗获取肺泡巨噬细胞(AM),检测支气管肺泡灌洗液(BALF)中肿瘤坏死因子-α(TNF-α)和白细胞介素1β(IL-1β)水平,同时行肺组织病理学检查.蛋白免疫印迹法测定各组AM中CYLD及NF-κB活性水平.结果 ANP大鼠肺损伤随着病情进展而逐渐加重.AM分泌TNF-α、IL-1β水平逐渐升高,至6h达到高峰,分别为(491.3±20.3)ng/L和(178.83±11.32) ng/L,12 h又回落.ANP组TNF-α及IL-1β含量较假手术组明显升高(P＜0.05),而GdCl3治疗组较ANP组明显降低(P＜0.05).NF-κB蛋白在假手术组表达呈低水平,而CYLD蛋白表达呈高水平.ANP组NF-κB蛋白表达从3h开始与假手术组相比已有上升,差异有统计学意义(P＜0.05),并随时间进行,表达逐渐增高.ANP组CYLD蛋白表达则相反,与假手术组相比从3h开始已有下降(P＜0.05),随时间进行,表达逐渐减少.GdCl3治疗组CYLD及NF-κB蛋白表达变化趋势与ANP组趋势相近,CYLD蛋白表达与ANP组相比,从6h起表达量开始增加(P＜0.05)；NF-κB蛋白表达则从1h起表达量明显下调,与ANP组相比差异有统计学意义(P＜0.05).ANP组、GdCl3治疗组AM中NF-κB与CYLD的表达活性均呈负相关(r分别为-0.918,-0.723,P＜0.01).结论 CYLD在ANP肺损伤中呈低度表达,且具有明显时相,从3h起表达逐渐减低；而NF-κB则相反,随时间进展,表达逐渐增高.经GdCl3处理后,CYLD表达增强,NF-κB表达降低,表明在ANP时CYLD的表达对NF-κB表达也有负性调控作用,进而使肺损伤减轻.
目的 研究腫瘤抑製蛋白CYLD在急性壞死性胰腺炎(ANP)肺損傷中的錶達狀況及其對覈轉錄因子-κB(NF-κB)的影響,探討其在肺損傷中的作用.方法 72隻成年SD大鼠隨機分成假手術組、ANP組及氯化釓(GdC13)治療組,每組24隻.採用逆行胰膽管註射5％牛磺痠鈉的方法建立ANP大鼠模型.各組術後1、3、6、12 h分批處死動物.經支氣管肺泡灌洗穫取肺泡巨噬細胞(AM),檢測支氣管肺泡灌洗液(BALF)中腫瘤壞死因子-α(TNF-α)和白細胞介素1β(IL-1β)水平,同時行肺組織病理學檢查.蛋白免疫印跡法測定各組AM中CYLD及NF-κB活性水平.結果 ANP大鼠肺損傷隨著病情進展而逐漸加重.AM分泌TNF-α、IL-1β水平逐漸升高,至6h達到高峰,分彆為(491.3±20.3)ng/L和(178.83±11.32) ng/L,12 h又迴落.ANP組TNF-α及IL-1β含量較假手術組明顯升高(P＜0.05),而GdCl3治療組較ANP組明顯降低(P＜0.05).NF-κB蛋白在假手術組錶達呈低水平,而CYLD蛋白錶達呈高水平.ANP組NF-κB蛋白錶達從3h開始與假手術組相比已有上升,差異有統計學意義(P＜0.05),併隨時間進行,錶達逐漸增高.ANP組CYLD蛋白錶達則相反,與假手術組相比從3h開始已有下降(P＜0.05),隨時間進行,錶達逐漸減少.GdCl3治療組CYLD及NF-κB蛋白錶達變化趨勢與ANP組趨勢相近,CYLD蛋白錶達與ANP組相比,從6h起錶達量開始增加(P＜0.05)；NF-κB蛋白錶達則從1h起錶達量明顯下調,與ANP組相比差異有統計學意義(P＜0.05).ANP組、GdCl3治療組AM中NF-κB與CYLD的錶達活性均呈負相關(r分彆為-0.918,-0.723,P＜0.01).結論 CYLD在ANP肺損傷中呈低度錶達,且具有明顯時相,從3h起錶達逐漸減低；而NF-κB則相反,隨時間進展,錶達逐漸增高.經GdCl3處理後,CYLD錶達增彊,NF-κB錶達降低,錶明在ANP時CYLD的錶達對NF-κB錶達也有負性調控作用,進而使肺損傷減輕.
목적 연구종류억제단백CYLD재급성배사성이선염(ANP)폐손상중적표체상황급기대핵전록인자-κB(NF-κB)적영향,탐토기재폐손상중적작용.방법 72지성년SD대서수궤분성가수술조、ANP조급록화구(GdC13)치료조,매조24지.채용역행이담관주사5％우광산납적방법건립ANP대서모형.각조술후1、3、6、12 h분비처사동물.경지기관폐포관세획취폐포거서세포(AM),검측지기관폐포관세액(BALF)중종류배사인자-α(TNF-α)화백세포개소1β(IL-1β)수평,동시행폐조직병이학검사.단백면역인적법측정각조AM중CYLD급NF-κB활성수평.결과 ANP대서폐손상수착병정진전이축점가중.AM분비TNF-α、IL-1β수평축점승고,지6h체도고봉,분별위(491.3±20.3)ng/L화(178.83±11.32) ng/L,12 h우회락.ANP조TNF-α급IL-1β함량교가수술조명현승고(P＜0.05),이GdCl3치료조교ANP조명현강저(P＜0.05).NF-κB단백재가수술조표체정저수평,이CYLD단백표체정고수평.ANP조NF-κB단백표체종3h개시여가수술조상비이유상승,차이유통계학의의(P＜0.05),병수시간진행,표체축점증고.ANP조CYLD단백표체칙상반,여가수술조상비종3h개시이유하강(P＜0.05),수시간진행,표체축점감소.GdCl3치료조CYLD급NF-κB단백표체변화추세여ANP조추세상근,CYLD단백표체여ANP조상비,종6h기표체량개시증가(P＜0.05)；NF-κB단백표체칙종1h기표체량명현하조,여ANP조상비차이유통계학의의(P＜0.05).ANP조、GdCl3치료조AM중NF-κB여CYLD적표체활성균정부상관(r분별위-0.918,-0.723,P＜0.01).결론 CYLD재ANP폐손상중정저도표체,차구유명현시상,종3h기표체축점감저；이NF-κB칙상반,수시간진전,표체축점증고.경GdCl3처리후,CYLD표체증강,NF-κB표체강저,표명재ANP시CYLD적표체대NF-κB표체야유부성조공작용,진이사폐손상감경.
Objective Acute necrotizing pancreatitis (ANP) may cause lung injury.This study explores two factors that are associated with lung damage from ANP,the expression of tumor suppressor factor CYLD and nuclear factor-kappa B (NF-κB).Methods 72 adult Sprague-Dawley rats were randomly divided into 3 groups:sham operation,ANP,and GdCl3 treatment groups (n=24 for each group).A retrograde injection of 5％ sodium taurocholate into the biliopancreeatic duct of rats induced ANP,and the animals were killed 1,3,6,and 12 hours after the ANP induction.AMs were harvested by bronchoalveolar lavage technique,and TNF-a and IL-1β levels in bronchoalveolar lavage fluid (BALF) were evaluated.Lung tissue was checked with histological examinations,and the activity of NF-κB and CYLD in AM were measured by western blot.Results TNF-α and IL-1β secreted by AM were gradually elevated,peaked on the sixth hour,had maximums of (491.3 ±20.3)ng/L and (178.83±11.32)ng/L respectively,and decreased on the twelfth hour.The levels of TNF-α and IL-1β in the ANP group were significantly higher than the sham operation group (P＜0.05),and the GdC13 group levels were obviously lower than ANP group.In the sham operation group,the expression of NF-κB was low and CYLD was high.In the ANP group,when compared to the sham operation group,the expression of NF-κB rose after 3 hours and continued to rise with time progression (P＜0.05).In contrast,CYLD protein expression in the ANP group dropped after 3 hours and continued to gradually decrease (P＜0.05).The CYLD and NF-κB protein expression in GdCl3 groups had similar trends as the ANP group.GdCl3 group CYLD levels began to rise at 6 hours (P＜0.05),and NF κB levels began to fall at 1 hour (P＜0.05).The expression of NF-κB and CYLD possessed a negative correlation in both the ANP and GdCl3 groups (r =-0.918,r=-0.723,P＜ 0.01).Conclusions Therefore,in acute lung injury associated with acute pancreatitis,CYLD expression decreased with evident phases,such as a decrease in levels after 3 hours,and NF κB expression increased.Also,GdCl3 may be responsible for upregulation of CYLD expression and downregulation of NF-κB expression,and confirmed that CYLD had a negative effect on NF-κB.Perhaps GdCl3 could be used in the future to ameliorate the lung injury associated with ANP.
