CAJ | 학술논문

目的：建立改良液氮冻伤法制作动脉粥样硬化（ AS）易损斑块（ VP）模型，验证改良法是否优于传统的造模方法。方法雄性新西兰白兔24只随机分为改良方法组（ A组）和传统方法组（ B组）各12只。分别按照改良方法及传统方法造模，6 w末各组随机抽取4只兔子（ A1组＝4只，B1组＝4只）检测斑块形成状况。 A组6 w末、B组10 w末剩余兔子（ A2组＝8只，B2组＝8只）以液氮激发斑块破裂。激发48 h后取出右颈总动脉行HE染色及免疫组化检测斑块内基质金属蛋白（MMP）－1水平。不同时间点分别采集空腹静脉血检测兔血脂水平总胆固醇（TC），甘油三酯（TG）及低密度脂蛋白（LDL）及血清超敏C反应蛋白（hs－CRP）水平。结果高脂3 d后兔血脂水平已明显升高，1 w后达基础水平的4～15倍；血脂变化曲线可见第1周血脂上升最为明显。与基线水平相比，高脂喂养1 w后两组兔血清hs－CRP即明显升高，至激发前进一步升高，激发后hs－CRP较激发前明显升高；各时间点两组间比较无显著性差异。大体观察及HE染色可见6 w末A1组兔已形成明显的AS化斑块，而B1组兔斑块较小；激发后48 h两组兔子的右颈总动脉均可见AS表面内皮损伤，部分形成血栓；两组易损斑块在组织结构上无明显差异，免疫组化染色见两组斑块处均有MMP－1蛋白的大量表达。结论改良液氮冻伤法可构建易损／破裂粥样硬化斑块，较传统方法明显缩短成模时间，节约造模成本，更快速，更高效。
목적：건립개량액담동상법제작동맥죽양경화（ AS）역손반괴（ VP）모형，험증개량법시부우우전통적조모방법。방법웅성신서란백토24지수궤분위개량방법조（ A조）화전통방법조（ B조）각12지。분별안조개량방법급전통방법조모，6 w말각조수궤추취4지토자（ A1조＝4지，B1조＝4지）검측반괴형성상황。 A조6 w말、B조10 w말잉여토자（ A2조＝8지，B2조＝8지）이액담격발반괴파렬。격발48 h후취출우경총동맥행HE염색급면역조화검측반괴내기질금속단백（MMP）－1수평。불동시간점분별채집공복정맥혈검측토혈지수평총담고순（TC），감유삼지（TG）급저밀도지단백（LDL）급혈청초민C반응단백（hs－CRP）수평。결과고지3 d후토혈지수평이명현승고，1 w후체기출수평적4～15배；혈지변화곡선가견제1주혈지상승최위명현。여기선수평상비，고지위양1 w후량조토혈청hs－CRP즉명현승고，지격발전진일보승고，격발후hs－CRP교격발전명현승고；각시간점량조간비교무현저성차이。대체관찰급HE염색가견6 w말A1조토이형성명현적AS화반괴，이B1조토반괴교소；격발후48 h량조토자적우경총동맥균가견AS표면내피손상，부분형성혈전；량조역손반괴재조직결구상무명현차이，면역조화염색견량조반괴처균유MMP－1단백적대량표체。결론개량액담동상법가구건역손／파렬죽양경화반괴，교전통방법명현축단성모시간，절약조모성본，경쾌속，경고효。
Objective To establish an improved high-fat diet combined with liquid nitrogen frostbite method to make vulnerable ath-erosclerotic plaque model and compare with the conventional liquid nitrogen frostbite method , verify that the modified method is better than the traditional modeling method .Methods 24 male New Zealand white rabbits were randomly divided into two groups:improved method of group (Group A =12) and the traditional method group (Group B =12).Rabbits in Group A were first fed with high fat diet a week , and then implementation of the right common carotid artery intima liquid nitrogen frostbite surgery , continued a high-fat diet for five weeks;the rabbits in Group B were firstly accepted the common carotid artery intima liquid nitrogen frostbite surgery , then a postoperative high fat diet for 10 weeks.At 6 weekend, 4 rabbits were randomly selected from two groups (Group A1 =4 the Group B1 =4), taken the right common carotid artery with gross observation and HE staining was used to detect plaque formation conditions .The remaining rabbits in Group A ac-cepted the second liquid nitrogen frostbite surgery to stimulate plaque rupture at six weekends and the ones in Group B at 10 weekend ( Group A2 =8 and Group B2 =8).All animals were sacrificed 48 h after the second surgery , removed the right common carotid artery to undergo HE staining and immunohistochemistry to detect MMP 1 level.Fasting venous blood was collected respectively at different time points to detect serum lipid levels ( TC, TG and LDL).And serum hs-CRP levels were detected.Results The lipid levels had significantly been increased after 3 days'high-fat diet, and 4~15 times more than the foundation level at 1 weekend.Rabbits of A1 group had formed obvious atheroscle-rotic plaque after 6 weeks'high-fat diet, but the plaques in B1 group were smaller.The serum hs-CRP was significantly elevated after one week high fat diet , and increased further with time .After the second surgery , hs-CRP was significantly increased than before .But these two groups showed no significant difference in each point time .48 h after the second surgery , HE staining showed endothelial injury of AS plaque surface and some thrombosis of two groups;there was no significant difference between the two groups of vulnerable plaque in the organiza -tional structure.Immunohistochemistry showed that MMP-1 protein expression was similar and mass in two groups .Conclusions The im-proved law could build vulnerable atherosclerotic plaques , shorter modeling time, saving the cost, be faster and more efficient.