CAJ | 학술논문

目的：观察大蒜素对高胆固醇饮食诱导的 apoE-/-小鼠动脉粥样硬化进展的影响,并从蛋白亚硝基化方面探讨其可能的作用机制。方法30只 apoE-/-小鼠随机分3组：对照组(生理盐水,ig)、低剂量组(大蒜素,9 mg/kg·d, ig)、高剂量组(大蒜素,18 mg/kg·d,ig)。高胆固醇饮食喂养12周,分别于第0、4、8、12周采血检测血脂水平,处理结束后,检测血浆氧化低密度脂蛋白(ox-LDL)、丙二醛(MDA)、肿瘤坏死因子-α(TNF-α)和一氧化氮(NO)水平；利用苏木精伊红染色与范吉尔森弹性纤维染色观察主动脉根部组织学变化；免疫组化染色检测动脉粥样硬化斑块内巨噬细胞和血管平滑肌细胞含量；免疫荧光观察主动脉根部蛋白亚硝基化水平的变化。另外,以人脐静脉内皮细胞为体外模型,将其与 ox-LDL(50μg/mL)共同培养24 h,并用不同剂量(10μmol/L 和20μmol/L)大蒜素加以处理,分别采用硝酸还原酶法和免疫荧光法检测细胞上清 NO 及细胞蛋白亚硝基化水平。结果组织学分析发现,大蒜素组小鼠主动脉根部斑块面积及斑块内巨噬细胞和血管平滑肌细胞含量明显较对照组少(P <0.05)。与对照组相比,大蒜素组小鼠血浆 MDA、ox-LDL、TNF-α水平明显降低(P <0.05),血浆 NO 水平及主动脉根部蛋白亚硝基化水平明显升高(P <0.05),但对其血脂水平无明显影响。体外实验结果显示,与对照组相比,大蒜素能够显著恢复由 ox-LDL 所导致的人脐静脉内皮细胞的 NO 和蛋白亚硝基化水平的减少(P <0.01)。结论大蒜素能够有效抑制动脉粥样硬化的进展,其作用机制可能与大蒜素通过上调内皮细胞蛋白亚硝基化水平所发挥抗氧化应激和抗炎作用有关。
목적：관찰대산소대고담고순음식유도적 apoE-/-소서동맥죽양경화진전적영향,병종단백아초기화방면탐토기가능적작용궤제。방법30지 apoE-/-소서수궤분3조：대조조(생리염수,ig)、저제량조(대산소,9 mg/kg·d, ig)、고제량조(대산소,18 mg/kg·d,ig)。고담고순음식위양12주,분별우제0、4、8、12주채혈검측혈지수평,처리결속후,검측혈장양화저밀도지단백(ox-LDL)、병이철(MDA)、종류배사인자-α(TNF-α)화일양화담(NO)수평；이용소목정이홍염색여범길이삼탄성섬유염색관찰주동맥근부조직학변화；면역조화염색검측동맥죽양경화반괴내거서세포화혈관평활기세포함량；면역형광관찰주동맥근부단백아초기화수평적변화。령외,이인제정맥내피세포위체외모형,장기여 ox-LDL(50μg/mL)공동배양24 h,병용불동제량(10μmol/L 화20μmol/L)대산소가이처리,분별채용초산환원매법화면역형광법검측세포상청 NO 급세포단백아초기화수평。결과조직학분석발현,대산소조소서주동맥근부반괴면적급반괴내거서세포화혈관평활기세포함량명현교대조조소(P <0.05)。여대조조상비,대산소조소서혈장 MDA、ox-LDL、TNF-α수평명현강저(P <0.05),혈장 NO 수평급주동맥근부단백아초기화수평명현승고(P <0.05),단대기혈지수평무명현영향。체외실험결과현시,여대조조상비,대산소능구현저회복유 ox-LDL 소도치적인제정맥내피세포적 NO 화단백아초기화수평적감소(P <0.01)。결론대산소능구유효억제동맥죽양경화적진전,기작용궤제가능여대산소통과상조내피세포단백아초기화수평소발휘항양화응격화항염작용유관。
Objective To investigate the effect of allicin on the development of atherosclerosis in apoE-/-mice and explore its underlying mechanism from the perspective of protein S-nitrosylation.Methods Thirty male apoE-/- mice were randomly divided into 3 groups:control group (saline,ig),low-dose group (allicin,9 mg/kg·d, ig)and high-dose group (allicin,18 mg/kg·d,ig).They were fed with high cholesterol diet for 12 weeks.The levels of plasma lipids,oxidized-LDL (ox-LDL),malondialdehyde,tumor necrosis factor-alpha and nitric oxide (NO)were measured.The atherosclerotic lesions in aortic root were evaluated after hematoxylin and eosin staining and elastica van Gieson and immunohistochemical staining,respectively.Furthermore,in vitro experiments were performed using human umbilical vein endothelial cells (HUVECs).The HUVECs were treated with allicin (10μmol/L or 20 μmol/L)for 24 hours in the presence of ox-LDL (50 μg/mL).The level of NO in supernatant was measured by a nitrate/nitrite assay. The protein S-nitrosylation of the HUVECs was detected through immunofluorescence.Results The histological analysis revealed that allicin treatment not only significantly decreased the areas of the atherosclerotic lesion (all P <0.05)but also suppressed the macrophage accumulation and smooth muscle cell proliferation in the lesion.There was no significant difference in the levels of plasma lipids between control and treated groups.However,allicin exerted obvious anti-oxidative and anti-inflammatory effects. Interestingly,the allicin treatment led to marked increase of the plasma NO level (P <0.05)and aortic protein S-nitrosylation.The experiments in vitro further proved that the allicin up-regulated the levels of NO and protein S-nitrosylation in HUVECs treated with ox-LDL (P < 0.01 ).Conclusion Allicin can inhibit the development of atherosclerosis.The mechanism is associated with the up-regulation of protein S-nitrosylation in endothelial cells, which plays an important role in anti-oxidization and anti-inflammation.