中南大学学报(医学版)
中南大學學報(醫學版)
중남대학학보(의학판)
JOURNAL OF CENTRAL SOUTH UNIVERSITY (MEDICAL SCIENCES)
2014年
11期
1137-1144
,共8页
肝%血液供给%缺血再灌注损伤%米诺环素%Wnt/β-catenin信号转导通路
肝%血液供給%缺血再灌註損傷%米諾環素%Wnt/β-catenin信號轉導通路
간%혈액공급%결혈재관주손상%미낙배소%Wnt/β-catenin신호전도통로
liver%blood supply%ischemia reperfusion injury%minocycline%Wnt /β-catenin signaling pathway
目的:探讨米诺环素对大鼠肝缺血再灌注损伤的保护作用及其分子机制。方法:54只雄性SD大鼠随机分成假手术对照组、缺血再灌注组和米诺环素组,每组18只。米诺环素组在缺血再灌注模型的基础上,自手术前36 h开始给予米诺环素灌胃给药(45 mg/kg),以后每12 h给药1次(22.5 mg/kg)。再灌注后2,6,24 h检测血清谷丙转氨酶(alanine aminotransferase,ALT)、谷草转氨酶(aspartate aminotransferase,AST)、乳酸脱氢酶(lactate dehydrogenase, LDH)水平;肝组织行HE染色,观察病理组织学改变并计算评分(根据Suzuki标准);肝组织匀浆检测丙二醛(malondialdehyde,MDA)、髓过氧化物酶(myeloperoxidase,MPO)水平;real-time PCR检测肝组织肿瘤坏死因子α(tumor necrosis factor-α,TNF-α)、白介素1β(interleukin-1 beta,IL-1β) mRNA的表达;Western印迹测定Dickkopf-1(DKK-1)和β-连环蛋白(β-catenin)的表达。结果:再灌注2,6,24 h后,与缺血再灌注组比较,米诺环素处理组血清ALT,AST,LDH水平,以及肝组织病理学半定量评分、肝组织中TNF-α和IL-1β mRNA水平的表达均降低(均P<0.05);氧化损伤指标MDA和MPO在肝组织中的含量减少(均P<0.05);米诺环素组DKK-1的蛋白表达下调(P<0.05),β-catenin蛋白表达上调(P<0.05)。结论:米诺环素可减轻缺血再灌注对肝的损伤,其机制可能是通过激活Wnt/β-catenin信号通路而减轻肝氧化应激,并抑制促炎性细胞因子的释放。
目的:探討米諾環素對大鼠肝缺血再灌註損傷的保護作用及其分子機製。方法:54隻雄性SD大鼠隨機分成假手術對照組、缺血再灌註組和米諾環素組,每組18隻。米諾環素組在缺血再灌註模型的基礎上,自手術前36 h開始給予米諾環素灌胃給藥(45 mg/kg),以後每12 h給藥1次(22.5 mg/kg)。再灌註後2,6,24 h檢測血清穀丙轉氨酶(alanine aminotransferase,ALT)、穀草轉氨酶(aspartate aminotransferase,AST)、乳痠脫氫酶(lactate dehydrogenase, LDH)水平;肝組織行HE染色,觀察病理組織學改變併計算評分(根據Suzuki標準);肝組織勻漿檢測丙二醛(malondialdehyde,MDA)、髓過氧化物酶(myeloperoxidase,MPO)水平;real-time PCR檢測肝組織腫瘤壞死因子α(tumor necrosis factor-α,TNF-α)、白介素1β(interleukin-1 beta,IL-1β) mRNA的錶達;Western印跡測定Dickkopf-1(DKK-1)和β-連環蛋白(β-catenin)的錶達。結果:再灌註2,6,24 h後,與缺血再灌註組比較,米諾環素處理組血清ALT,AST,LDH水平,以及肝組織病理學半定量評分、肝組織中TNF-α和IL-1β mRNA水平的錶達均降低(均P<0.05);氧化損傷指標MDA和MPO在肝組織中的含量減少(均P<0.05);米諾環素組DKK-1的蛋白錶達下調(P<0.05),β-catenin蛋白錶達上調(P<0.05)。結論:米諾環素可減輕缺血再灌註對肝的損傷,其機製可能是通過激活Wnt/β-catenin信號通路而減輕肝氧化應激,併抑製促炎性細胞因子的釋放。
목적:탐토미낙배소대대서간결혈재관주손상적보호작용급기분자궤제。방법:54지웅성SD대서수궤분성가수술대조조、결혈재관주조화미낙배소조,매조18지。미낙배소조재결혈재관주모형적기출상,자수술전36 h개시급여미낙배소관위급약(45 mg/kg),이후매12 h급약1차(22.5 mg/kg)。재관주후2,6,24 h검측혈청곡병전안매(alanine aminotransferase,ALT)、곡초전안매(aspartate aminotransferase,AST)、유산탈경매(lactate dehydrogenase, LDH)수평;간조직행HE염색,관찰병리조직학개변병계산평분(근거Suzuki표준);간조직균장검측병이철(malondialdehyde,MDA)、수과양화물매(myeloperoxidase,MPO)수평;real-time PCR검측간조직종류배사인자α(tumor necrosis factor-α,TNF-α)、백개소1β(interleukin-1 beta,IL-1β) mRNA적표체;Western인적측정Dickkopf-1(DKK-1)화β-련배단백(β-catenin)적표체。결과:재관주2,6,24 h후,여결혈재관주조비교,미낙배소처리조혈청ALT,AST,LDH수평,이급간조직병이학반정량평분、간조직중TNF-α화IL-1β mRNA수평적표체균강저(균P<0.05);양화손상지표MDA화MPO재간조직중적함량감소(균P<0.05);미낙배소조DKK-1적단백표체하조(P<0.05),β-catenin단백표체상조(P<0.05)。결론:미낙배소가감경결혈재관주대간적손상,기궤제가능시통과격활Wnt/β-catenin신호통로이감경간양화응격,병억제촉염성세포인자적석방。
Objective: To explore the protective effect of minocycline on hepatic ischemia-reperfusion injury (IRI) in rats and the underlying mechanisms. Methods: A total of 54 male Sprague-Dawley rats were randomly divided into 3 groups: the sham-operated group (control group), the ischemic–reperfusion (IR group), and the minocycline preconditioning group (n=18 per group). The rats in the minocycline preconditioning group were given minocycline (45 mg/kg) by gastric irrigation at 36 h before operation and then were subsequently administered with minocycline (22.5 mg/kg) at every 12 h. hTe rats were sacriifcedat 2, 6, 24 h after reperfusion. The serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST) and lactate dehydrogenase (LDH) were measured. HE staining of liver tissues was performed to detect the histological changes, and the degree of liver IRI according to Suzuki score were calculated. hTe levels of malondialdehyde (MDA) and myeloperoxidase (MPO) were determined by spectrophotometer; the mRNA expression of tumor necrosis factor-α (TNF-α) and interleukin-1 beta (IL-1β) in the liver were measured by real-time PCR; Dickkopf-1 (DKK-1) and beta-catenin (β-catenin) protein expression in the liver were detected by Western blot. Results: Atfer 2, 6, 24 h reperfusion, compared with the IR group, the liver function (ALT, AST and LDH) in the minocycline group was signiifcantly improved (allP<0.05); the Suzuki’s scores and the levels of hepatic TNF-α and IL-1β mRNA were signiifcantly decreased (allP<0. 05); the MDA and MPO levels the liver were decreased (bothP<0.05); the protein expression of hepatic DKK-1 was decreased (P<0.05), while the protein expression of β-catenin was increased (P<0.05). Conclusion: Minocycline can alleviate the ischemic-reperfusion injury mainly through reducing oxidative stress and inhibiting the release of pro-inlfammatory cytokines depends on the activation of the Wnt/β-catenin signaling pathway in the liver.