CAJ | 학술논문

目的探讨CTLA-4Ig融合蛋白抑制高脂饮食饲养的载脂蛋白E基因缺陷小鼠(apoE-/-)动脉粥样硬化(AS)的作用机理.方法 30只10周龄apoE-/-小鼠,随机分为3组(每组10只),高脂饮食饲养,分别采用CTLA-4Ig、IgGI、PBS腹腔注射(每周2次).12周后取小鼠主动脉进行病理学检查,检测AS病变相关指标.包括斑块面积/管腔面积比、血管内膜/中膜厚度比,斑块内脂质、胶原和平滑肌细胞含量;取血清检测总胆固醇浓度和CRP、sICAM-1、IFN-γ、IL-10、TGF-β1浓度.结果高脂饮食饲养12周后,形成明显AS斑块,但CTLA-4Ig组病变最轻.CTLA-4Ig组、IgG1组和PBS组的斑块面积/管腔面积比、血管内膜/中膜厚度比、斑块内脂质含量和胶原含量差异均有统计学意义(P<0.05),CTLA-4Ig组斑块面积/管腔面积比、血管内膜/中膜厚度比和斑块内脂质含量均显著低于IgG1组和PBS组(P<0.05),其斑块内胶原含量高于IgG1组和PBS组(P<0.05),而后2组间上述指标的差异均无统计学意义(P>0.05);3组间斑块内平滑肌细胞含量差异不具有统计学意义(P>0.05).CTLA-4Ig组、IgG1组和PBS组的血清总胆固醇浓度差异不具有计学意义(P>0.05),3组的血清CRP、sICAM-1、IFN-γ、IL-10、TGF-β1水平差异均有统计学意义(P<0.05),CTLA-4Ig组血清CRP、sICAM-1、IFN-γ水平降低,血清IL-10、TGF-β1水平升高,而后2组间上述指标的差异均无统计学意义(P>0.05).结论 CTLA-4Ig融合蛋白抑制高脂饮食饲养的apoE(-/-)小鼠动脉粥样硬化进程,可能与阻断B7/CD28通路、抗炎作用、促进Th2极化、及影响调节性T细胞功能等有关.
목적 탐토CTLA-4Ig융합단백억제고지음식사양적재지단백E기인결함소서(apoE-/-)동맥죽양경화(AS)적작용궤리.방법 30지10주령apoE-/-소서,수궤분위3조(매조10지),고지음식사양,분별채용CTLA-4Ig、IgGI、PBS복강주사(매주2차).12주후취소서주동맥진행병이학검사,검측AS병변상관지표.포괄반괴면적/관강면적비、혈관내막/중막후도비,반괴내지질、효원화평활기세포함량;취혈청검측총담고순농도화CRP、sICAM-1、IFN-γ、IL-10、TGF-β1농도.결과고지음식사양12주후,형성명현AS반괴,단CTLA-4Ig조병변최경.CTLA-4Ig조、IgG1조화PBS조적반괴면적/관강면적비、혈관내막/중막후도비、반괴내지질함량화효원함량차이균유통계학의의(P<0.05),CTLA-4Ig조반괴면적/관강면적비、혈관내막/중막후도비화반괴내지질함량균현저저우IgG1조화PBS조(P<0.05),기반괴내효원함량고우IgG1조화PBS조(P<0.05),이후2조간상술지표적차이균무통계학의의(P>0.05);3조간반괴내평활기세포함량차이불구유통계학의의(P>0.05).CTLA-4Ig조、IgG1조화PBS조적혈청총담고순농도차이불구유계학의의(P>0.05),3조적혈청CRP、sICAM-1、IFN-γ、IL-10、TGF-β1수평차이균유통계학의의(P<0.05),CTLA-4Ig조혈청CRP、sICAM-1、IFN-γ수평강저,혈청IL-10、TGF-β1수평승고,이후2조간상술지표적차이균무통계학의의(P>0.05).결론 CTLA-4Ig융합단백억제고지음식사양적apoE(-/-)소서동맥죽양경화진정,가능여조단B7/CD28통로、항염작용、촉진Th2겁화、급영향조절성T세포공능등유관.
Objective To investigate the mechanisms of inhibitory effect of CTLA-4Ig fusion protein on athero-sclerosis in mice with apolipoprotein-E gene deficient fed on high cholesterol diet. Methods 30 male 10-week-old apoE(-/-) mice fed on cholesterol diet were randomly divided into CTLA-4Ig treatment group, IgG1 group and PBS group. There were 10 mice in each group. Mice in the three groups were given intraperitoneal injection of CTLA-41g (10 μg per time), Rat-IgG1 (10 μg per time), PBS (100 μL per time) respectively, twice a week. Followed by 12-week treatment, the whole aorta from the root to crotch of lilac artery was separated and sera were collected. Subsequently, the area ratio of plaque and lumen in cross section, the thiekness ratio of endangium and tuniea media, the intraplaque lipid-soaking extent and the amount of intraplaque collagen fibrils and smooth muscle cells were analyzed by image-processing. The concentrations of total cholesterol, CRP, sI-CAM-1, IFN-γ, IL-10 and TGF-β1 in the sera were measured. Results There were typical atherosclerotic plaques in apoE(-/-) mice fed on high cholesterol diet for 12 weeks. The plaque was mild in CTLA-4Ig group. There were significant difference in the area ratio of plaque and lumen, the thickness ratio of endangium and tu-nica media, the intraplaque lipid-soaking extent, and the amount of collagen fibrils among the three groups (all P <0.05). It was fnund that the area ratio of plaque and lumen, the thickness ratio of endangium and tunica media, and the intraplaque lipid-soaking extent were significantly lower and the amount of collagen fibrils was significantly higher in CTLA-4Ig group than those in IgG1 group and PBS group(all P <0.05), but there was no significant difference in those between the IgG1 group and PBS group (all P >0.05). There were no signifi-cant difference in the amount of smooth muscle cells in the three groups(P > 0. 05). There were no significant difference in the concentration of total cholesterol in the sera in the three groups (P > 0. 05). There was signifi-cant difference in the concentrations of CRP, sICAM-1, IFN-γ, IL-10, and TGF-β1 in the sera in the three groups (all P <0. 05). It was found that CTLA-4Ig could decrease the concentrations of CRP, sICAM-1 and IFN-γ and increase IL-10 and TGF-β1 in the sera, but IgG1 and PBS could not. Conclusion CTLA-4Ig fu-sion protein could inhibit the atherosclerosis progress in apoE(-/-) mice fed on high cholesterol diet and it' s effect might be associated with blocking B7/CD28, anti-inflammation, promoting Th2 polarization and affecting regulatory T cells.