微量元素与健康研究

目的 观察辛伐他汀可溶性微针对动脉粥样硬化(AS)小鼠脂质代谢、炎症因子的影响，探讨辛伐他汀可溶性微针对AS炎症的调控作用。方法 将6只C57BL/6J小鼠设为空白组，予以普通饲料喂养，18只ApoE^-/-小鼠以高脂饲料喂养建立AS模型，喂养8周后，随机分为模型组、辛伐他汀组、可溶性微针组，每组6只。空白组、模型组仅保持与治疗组同样的抓取手法，不予干预；辛伐他汀组予以辛伐他汀灌胃(2.5 mg/kg),每日1次，连续8周；可溶性微针组给予与灌胃剂量相同的剂量辛伐他汀可溶性微针贴片干预，隔天1次，连续干预8周。采用全自动生化分析仪测定小鼠血清中总胆固醇(total cholesterol, TC)、甘油三酯(triglyceride, TG)、低密度脂蛋白胆固醇(low density lipoprotein cholesterol, LDL-C)、高密度脂蛋白胆固醇(high-density lipoprotein cholesterol, HDL-C)含量；HE染色法观察小鼠主动脉弓病理形态；酶联免疫吸附测定法(ELISA)检测小鼠血清中TNF-α、IL-1β含量；免疫组化法检测小鼠主动脉中TNF-α、IL-1β蛋白表达情况。结果 与空白组比较，模型组小鼠血清中TC、TG、LDL-C含量均显著升高(P<0.01),HDL-C含量显著降低(P<0.01),小鼠主动脉弓组织见血管内膜损伤，血管内皮细胞脱落，且部分组织可见局部纤维断裂、溶解，排列紊乱，小鼠血清中TNF-α、IL-1β含量显著升高(P<0.01),主动脉组织中TNF-α、IL-1β蛋白表达着色较深，以棕黄色、褐色颗粒为主；与模型组比较，辛伐他汀组小鼠血清中TC、TG、LDL-C含量均显著降低(P<0.01),HDL-C含量显著升高(P<0.01),小鼠主动脉弓血管管腔结构较为完整，内膜、中膜、外膜结构清晰，内膜变性情况减少，小鼠血清中TNF-α、IL-1β含量显著下降(P<0.01),小鼠主动脉组织中TNF-α、IL-1β蛋白表达中棕黄色染色颗粒数量减少，染色强度减弱；可溶性微针组小鼠血清中TC、TG、LDL-C含量显著降低(P<0.01),HDL-C含量显著升高(P<0.01),小鼠主动脉弓内膜损伤减轻，可见少量内皮细胞脱落，小鼠血清中TNF-α、IL-1β含量均明显下降(P<0.01),小鼠主动脉组织中TNF-α、IL-1β蛋白染色程度变浅，以散在阳性表达为主。结论 辛伐他汀可溶性微针可改善AS模型ApoE^-/-小鼠脂质代谢水平，改善主动脉弓病理损伤，抑制TNF-α、IL-1β促炎因子的表达，对动脉粥样硬化小鼠炎症具有一定的抑制作用。

Objective To investigate the effects of simvastatin-loaded dissolving microneedles on lipid metabolism and inflammatory factors in mice with atherosclerosis(AS) and to evaluate their potential regulatory effects on AS-associated inflammation.Methods Six C57BL/6J mice were assigned to the control group and fed a standard diet, while 18 ApoE^-/- mice were fed a high-fat diet to establish an AS model. After 8 weeks, the ApoE^-/- mice were randomly assigned to the model, simvastatin, and dissolving microneedle groups(n=6 per group). Mice in the control and model groups received no intervention other than handling identical to that of the treatment groups. Mice in the simvastatin group received simvastatin by oral gavage at 2.5 mg/kg once daily for 8 consecutive weeks. Mice in the dissolving microneedle group were treated with simvastatin-loaded dissolving microneedle patches at a dose equivalent to that administered by gavage, once every other day for 8 weeks. Serum levels of total cholesterol(TC), triglycerides(TG), low-density lipoprotein cholesterol(LDL-C), and high-density lipoprotein cholesterol(HDL-C) were measured using an automated biochemical analyzer. Histopathological changes in the aortic arch were evaluated by hematoxylin and eosin(H&E) staining. Serum levels of tumor necrosis factor-α(TNF-α) and interleukin-1β(IL-1β) were determined by enzyme-linked immunosorbent assay(ELISA), and the protein expression of TNF-α and IL-1β in aortic tissues was assessed by immunohistochemistry.Results Compared with the control group, mice in the model group exhibited significantly increased serum levels of TC, TG, and LDL-C and significantly decreased HDL-C levels(P<0.01). Histopathological examination revealed intimal injury in the aortic arch, endothelial cell detachment, and focal fiber disruption and dissolution with disorganized tissue architecture. Serum TNF-α and IL-1β levels were significantly elevated(P<0.01), and immunohistochemical staining for TNF-α and IL-1β in aortic tissues was markedly enhanced, predominantly appearing as yellow-brown to brown granules. Compared with the model group, the simvastatin group showed significantly decreased serum TC, TG, and LDL-C levels and increased HDL-C levels(P<0.01). The structure of the aortic lumen was relatively well preserved, with clearly distinguishable intimal, medial, and adventitial layers and reduced intimal degeneration. Serum TNF-α and IL-1β levels were significantly decreased(P<0.01), accompanied by fewer yellow-brown TNF-α-and IL-1β-positive granules and reduced staining intensity in aortic tissues. Similarly, mice in the dissolving microneedle group exhibited significantly decreased serum TC, TG, and LDL-C levels and increased HDL-C levels(P<0.01). Intimal injury in the aortic arch was attenuated, with only limited endothelial cell detachment. Serum TNF-α and IL-1β levels were also significantly reduced(P<0.01), while immunohistochemical staining for TNF-α and IL-1β in aortic tissues was weaker and predominantly showed scattered positive expression.Conclusion Simvastatin-loaded dissolving microneedles improve lipid metabolism and alleviate pathological injury of the aortic arch in ApoE^-/- mice with AS while suppressing the expression of the pro-inflammatory factors TNF-α and IL-1β. These findings suggest that simvastatin-loaded dissolving microneedles exert anti-inflammatory effects in atherosclerotic mice.