CAJ | 학술논문

目的观察线粒体通透性转换孔(mPTP)在缺血后适应减轻兔小肠缺血再灌注(I/R)损伤中的作用.方法将新西兰兔40只随机分为假手术组(Sham组)、L/R组、缺血后适应组(IPO组)、mPTP抑制剂环孢素A组(CsA组)、IPO+ mPTP开放剂苍术甙组(IPO+ Atr组).分别干预后采集各组兔部分小肠组织标本,苏木素-伊红(HE)染色,检测小肠组织丙二醛(MDA)活性,提取肠线粒体,检测mPTP开放,Chiu氏6级评分法观察肠黏膜损伤,原位缺口末端标记法(TUNEL)法检测肠上皮细胞凋亡.结果与Sham组比较,I/R组mPTP开放明显增加(I/R组3.53±0.36比Sham组1.37 ±0.16,P＜ 0.05);MDA活性明显增高[I/R组(0.98±0.14) nmol/mg比Sham组(0.34±0.03) nmol/mg,P＜0.05];肠黏膜损伤评分明显增高[I/R组(4.66±0.41)分比Sham组(0.92±0.58)分,P＜0.05];肠细胞凋亡指数明显增高[I/R组(60.34±6.02)％比Sham组(4.65±1.68)％,P＜0.05].与I/R组比较,IPO组和CsA组mPTP开放明显减少(IPO组2.32 ±0.23、CsA组2.62±0.18比I/R组3.53 ±0.36,P＜0.05);MDA活性明显减低[IPO组(0.55 ±0.04) nmol/mg、CsA组(0.62±0.06) nmol/mg比I/R组(0.98 ±0.14) nmol/mg,P＜0.05];肠黏膜损伤评分明显减低[IPO组(3.25±0.27)分、CsA组(3.52 ±0.55)分比I/R组(4.66 ±0.41)分,P＜0.05];肠细胞凋亡指数明显减低[IPO组(28.33±3.20)％、CsA组(20.49±4.10)％比I/R组(60.34±6.02)％,P＜0.05],与IPO组比较,IPO+ Atr组mPTP开放明显增加(IPO+ Atr组1.05±0.16比IPO组2.32±0.23,P＜0.05);MDA活性明显增高[IPO +Atr组(1.08±0.18) nmol/mg比IPO组(0.55±0.04) nmol/mg,P＜0.05];肠黏膜损伤评分明显增加[IPO+ Atr组(4.57±0.32)分比IPO组(3.25±0.27)分,P＜0.05];肠细胞凋亡指数明显增加[IPO+ Atr组(40.35±2.18)％比IPO组(28.33±3.20)％,P＜0.05].结论缺血后适应能减轻兔肠I/R损伤,其机制可能与抑制mPTP开放有关.
목적 관찰선립체통투성전환공(mPTP)재결혈후괄응감경토소장결혈재관주(I/R)손상중적작용.방법 장신서란토40지수궤분위가수술조(Sham조)、L/R조、결혈후괄응조(IPO조)、mPTP억제제배포소A조(CsA조)、IPO+ mPTP개방제창술대조(IPO+ Atr조).분별간예후채집각조토부분소장조직표본,소목소-이홍(HE)염색,검측소장조직병이철(MDA)활성,제취장선립체,검측mPTP개방,Chiu씨6급평분법관찰장점막손상,원위결구말단표기법(TUNEL)법검측장상피세포조망.결과 여Sham조비교,I/R조mPTP개방명현증가(I/R조3.53±0.36비Sham조1.37 ±0.16,P＜ 0.05);MDA활성명현증고[I/R조(0.98±0.14) nmol/mg비Sham조(0.34±0.03) nmol/mg,P＜0.05];장점막손상평분명현증고[I/R조(4.66±0.41)분비Sham조(0.92±0.58)분,P＜0.05];장세포조망지수명현증고[I/R조(60.34±6.02)％비Sham조(4.65±1.68)％,P＜0.05].여I/R조비교,IPO조화CsA조mPTP개방명현감소(IPO조2.32 ±0.23、CsA조2.62±0.18비I/R조3.53 ±0.36,P＜0.05);MDA활성명현감저[IPO조(0.55 ±0.04) nmol/mg、CsA조(0.62±0.06) nmol/mg비I/R조(0.98 ±0.14) nmol/mg,P＜0.05];장점막손상평분명현감저[IPO조(3.25±0.27)분、CsA조(3.52 ±0.55)분비I/R조(4.66 ±0.41)분,P＜0.05];장세포조망지수명현감저[IPO조(28.33±3.20)％、CsA조(20.49±4.10)％비I/R조(60.34±6.02)％,P＜0.05],여IPO조비교,IPO+ Atr조mPTP개방명현증가(IPO+ Atr조1.05±0.16비IPO조2.32±0.23,P＜0.05);MDA활성명현증고[IPO +Atr조(1.08±0.18) nmol/mg비IPO조(0.55±0.04) nmol/mg,P＜0.05];장점막손상평분명현증가[IPO+ Atr조(4.57±0.32)분비IPO조(3.25±0.27)분,P＜0.05];장세포조망지수명현증가[IPO+ Atr조(40.35±2.18)％비IPO조(28.33±3.20)％,P＜0.05].결론 결혈후괄응능감경토장I/R손상,기궤제가능여억제mPTP개방유관.
Objective To investigate the role of mitochondrial permeability transition pore (mPTP) in ischemic post-conditioning for counteracting small intestinal ischemia-reperfusion (I/R) injury in rabbits.Methods Forty New Zealand rabbits were randomly divided into sham-operated group (Sham group),I/R group,ischemic post-conditioning group (IPO group),mPTP inhibitor cyclosporine A group (CsA group),and IPO + mPTP agonist atraotydin group (IPO +Atr group).Parts of small intestine tissues were collected from each group of rabbits after intervention for hematoxylin-eosin (HE) staining.Malondialdehyde (MDA) activity in small intestine tissue was measured by the relevant kit.The mitochondria were isolated and mPTP opening was detected by relevant kits.Intestinal mucosal injury was observed by Chiu' s grade 6 scoring method.The apoptosis of intestinal epithelial cells was examined by TdT-mediated dUTP nick end labeling (TUNEL) assay.Results As compared with Sham group,mPTP opening in I/R group was significantly increased (3.53 ± 0.36 vs.1.37 ± 0.16,P ＜ 0.05) ; MDA activity was significantly increased [(0.98 ±0.14) nmol/mg vs.(0.34 ±0.03) nmol/mg,P＜0.05] ; intestinal mucosa score was significantly increased (4.66 ± 0.41 vs.0.92 ± 0.58,P ＜ 0.05) ; apoptosis index of intestinal cells was significantly increased [(60.34 ± 6.02) ％ vs.(4.65 ± 1.68) ％,P ＜ 0.05].As compared with I/R group,mPTP openings in IPO group and CsA group were significantly decreased (2.32 ± 0.23 and 2.62 ±0.18 vs.3.53 ±0.36,P ＜0.05) ; MDA activity was significantly decreased [(0.55 ± 0.04) and (0.62 ± 0.06) nmol/mg vs.(0.98 ± 0.14) nmol/mg,P ＜ 0.05] ; intestinal mucosa score was significantly decreased (3.25 ±0.27 and 3.52 ±0.55 vs.4.66 ±0.41,P ＜0.05) ; apoptosis of intestinal cells was significantly decreased [(28.33 ± 3.20) ％ and (20.49 ± 4.10) ％ vs.(60.34 ± 6.02) ％,P ＜ 0.05].As compared with IPO group,mPTP openings in IPo + Atr group were significantly increased (2.32 ± 0.23 vs.1.05 ± 0.16,P ＜ 0.05) ; MDA activity was significantly increased [(1.08 ± 0.18) vs.(0.55 ± 0.04) nmol/mg,P ＜ 0.05] ; intestinal mucosa score was significantly increased (4.57 ±0.32 vs.3.25 ±0.27,P ＜0.05) ; apoptosis index of intestinal cells was significantly increased [(40.35±2.18)％ vs.(28.33±3.20)％,P＜0.05].Conclusion The mechanism of ischemic post-conditioning for counteracting intestinal ischemia-reperfusion injury in rabbits may be associated with the inhibition of mitochondrial permeability transition pore opening.