中华器官移植杂志
中華器官移植雜誌
중화기관이식잡지
CHINESE JOURNAL OF ORGAN TRANSPLANTATION
2015年
3期
141-146
,共6页
汪笑宇%朱有华%赵闻雨%曾力
汪笑宇%硃有華%趙聞雨%曾力
왕소우%주유화%조문우%증력
缺血%再灌注损伤%抗原,CD11b%枯否细胞%肝%小鼠
缺血%再灌註損傷%抗原,CD11b%枯否細胞%肝%小鼠
결혈%재관주손상%항원,CD11b%고부세포%간%소서
Ischemia%Reperfusion injury%Antigens,CD11 b%Kupffer cells%Liver%Mice
目的 探讨整合素CD11b在小鼠肝脏缺血再灌注损伤中的作用及其可能的机制.方法 使用CD11b缺陷(CD11b-/-组)的C57BL/6小鼠和正常C57BL/6小鼠(WT组),制作小鼠70％肝脏缺血再灌注损伤模型:维持小鼠体温为37℃,暴露肝叶后用无损伤显微血管夹阻断肝脏左叶和中叶血流60 min,然后恢复肝脏左叶和中叶血流,最后缝合腹部切口;另设假手术组(CD11b-/-Sham组和WT Sham组)作为对照.再灌注后1、3、6、12、24和48 h取血液,检测血清丙氨酸转氨酶(ALT)和天冬氨酸转氨酶(AST)水平;肝组织样本行HE染色和凋亡检测;Trizol试剂盒一步法提取肝组织细胞总RNA,检测肿瘤坏死因子α(TNF-α)和白细胞介素10(IL-10)的表达情况;提取枯否细胞,检测还原性辅酶Ⅱ(NADPH)氧化酶活性及活性氧的生成情况.结果 CD11b-/-组小鼠术后ALT、AST升高程度显著低于WT组小鼠(P＜0.05).肝组织HE染色显示CD11b-/-组小鼠肝细胞坏死、充血和肿胀程度明显轻于WT组小鼠(P＜0.05);CD11b-/-组缺血再灌注后3h肝细胞凋亡比例为(22.8±5.8)％,低于WT组的(66.6±10.2)％,差异有统计学意义(P＜0.05).CD11b-/-组小鼠TNF-α表达低于WT组小鼠,而IL-10表达却高于WT组小鼠(P＜0.05).CD11b-/-组小鼠的NADPH氧化酶活性及活性氧产生均低于WT小鼠(P＜0.05).结论 整合素CD11b可以调控炎症因子(TNF-α)和抗炎症因子(IL-10)的释放,增强枯否细胞NADPH氧化酶的活性,继而影响活性氧的生成,最终加重肝脏缺血再灌注损伤.
目的 探討整閤素CD11b在小鼠肝髒缺血再灌註損傷中的作用及其可能的機製.方法 使用CD11b缺陷(CD11b-/-組)的C57BL/6小鼠和正常C57BL/6小鼠(WT組),製作小鼠70％肝髒缺血再灌註損傷模型:維持小鼠體溫為37℃,暴露肝葉後用無損傷顯微血管夾阻斷肝髒左葉和中葉血流60 min,然後恢複肝髒左葉和中葉血流,最後縫閤腹部切口;另設假手術組(CD11b-/-Sham組和WT Sham組)作為對照.再灌註後1、3、6、12、24和48 h取血液,檢測血清丙氨痠轉氨酶(ALT)和天鼕氨痠轉氨酶(AST)水平;肝組織樣本行HE染色和凋亡檢測;Trizol試劑盒一步法提取肝組織細胞總RNA,檢測腫瘤壞死因子α(TNF-α)和白細胞介素10(IL-10)的錶達情況;提取枯否細胞,檢測還原性輔酶Ⅱ(NADPH)氧化酶活性及活性氧的生成情況.結果 CD11b-/-組小鼠術後ALT、AST升高程度顯著低于WT組小鼠(P＜0.05).肝組織HE染色顯示CD11b-/-組小鼠肝細胞壞死、充血和腫脹程度明顯輕于WT組小鼠(P＜0.05);CD11b-/-組缺血再灌註後3h肝細胞凋亡比例為(22.8±5.8)％,低于WT組的(66.6±10.2)％,差異有統計學意義(P＜0.05).CD11b-/-組小鼠TNF-α錶達低于WT組小鼠,而IL-10錶達卻高于WT組小鼠(P＜0.05).CD11b-/-組小鼠的NADPH氧化酶活性及活性氧產生均低于WT小鼠(P＜0.05).結論 整閤素CD11b可以調控炎癥因子(TNF-α)和抗炎癥因子(IL-10)的釋放,增彊枯否細胞NADPH氧化酶的活性,繼而影響活性氧的生成,最終加重肝髒缺血再灌註損傷.
목적 탐토정합소CD11b재소서간장결혈재관주손상중적작용급기가능적궤제.방법 사용CD11b결함(CD11b-/-조)적C57BL/6소서화정상C57BL/6소서(WT조),제작소서70％간장결혈재관주손상모형:유지소서체온위37℃,폭로간협후용무손상현미혈관협조단간장좌협화중협혈류60 min,연후회복간장좌협화중협혈류,최후봉합복부절구;령설가수술조(CD11b-/-Sham조화WT Sham조)작위대조.재관주후1、3、6、12、24화48 h취혈액,검측혈청병안산전안매(ALT)화천동안산전안매(AST)수평;간조직양본행HE염색화조망검측;Trizol시제합일보법제취간조직세포총RNA,검측종류배사인자α(TNF-α)화백세포개소10(IL-10)적표체정황;제취고부세포,검측환원성보매Ⅱ(NADPH)양화매활성급활성양적생성정황.결과 CD11b-/-조소서술후ALT、AST승고정도현저저우WT조소서(P＜0.05).간조직HE염색현시CD11b-/-조소서간세포배사、충혈화종창정도명현경우WT조소서(P＜0.05);CD11b-/-조결혈재관주후3h간세포조망비례위(22.8±5.8)％,저우WT조적(66.6±10.2)％,차이유통계학의의(P＜0.05).CD11b-/-조소서TNF-α표체저우WT조소서,이IL-10표체각고우WT조소서(P＜0.05).CD11b-/-조소서적NADPH양화매활성급활성양산생균저우WT소서(P＜0.05).결론 정합소CD11b가이조공염증인자(TNF-α)화항염증인자(IL-10)적석방,증강고부세포NADPH양화매적활성,계이영향활성양적생성,최종가중간장결혈재관주손상.
Objective To investigate the role of integrin CD11b in liver ischemia/reperfusion (I/R) injury and its possible mechanism.Method CD11b-/-and WT (C57BL/6) mice were used to establish a 70％ liver warm I/R by clamping the left and median liver lobes for 60 min with vascular micro clamp at 37℃,then the clamp was removed and the abdominal incision was sutured.The blood plasma and liver samples were obtained at different time points (1,3,6,12,24 and 48 h) postreperfusion to assess liver function and cellular injury.Serum ALT and AST levels were determined,and HE staining and TUNEL assay were performed to estimate the severity of liver damage.Tumor necrosis factor-α (TNF-α) and interleukin-10 (IL-10) were assayed by Reverse transcription polymerase chain reaction (RT-PCR).Kupffer cells were isolated from the live,and the reduced form of nicotinamide-adenine dinucleotide phosphate (NADPH) oxidase activity and active oxygen species (ROS) production were assayed.Result CD11b-/-mice displayed a significantly preserved liver function as represented by lower alanine aminotransferase (ALT) and aspartic transaminase (AST) levels,less histological damage and apoptosis compared to WT mice.Furthermore,TNF-α was decreased and IL-10 mRNA expression was increased in CD11b-/-mice compared to WT mice.Finally,CD11b-/-mice showed decreased activity of NADPH oxidase and less ROS production.Conclusion Integrin CD11 b may regulate the levels of inflammatory (TNF-α) and anti-inflammatory (IL-10) cytokines,enhance the activity of NADPH oxidase in Kupffer cells and enrich the production of ROS,which aggravate liver I/R injury.
