首页|miR-126阳离子纳米乳的制备及初步药效

miR-126阳离子纳米乳的制备及初步药效

Preparation of miR-126 cationic nanoemulsion and its preliminary pharmacodynamic

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目的 采用阳离子纳米乳(CNE)载miRNA-126(miR-126)形成miR-126阳离子纳米乳递送系统(CNE-miR126),并对其药剂学性能、时间稳定性、血清稳定性、细胞毒性、细胞摄取及血管黏附分子-1(VCAM-1)蛋白表达量进行评估.方法 用琼脂糖凝胶电泳考察CNE和miR-126的最佳氮磷比(N/P),采用纳米粒度及Zeta电位分析测量仪测定其粒径和进行药剂学性能评价;在4 ℃条件下,评估CNE-miR-126在1周内粒径及电位的稳定性;用血清稳定性实验评估CNE-miR-126在48 h之内的稳定性,MTT法评估CNE-miR-126的细胞毒性;流式细胞术和荧光显微镜观察人脐静脉内皮细胞(HUVECs)对CNE-miR-126的摄取情况,免疫蛋白质印迹法(Weston blot)检测CNE-miR-126对由脂多糖(LPS)致炎的HUVECs中VCAM-1的表达水平.结果 CNE与miR-126 最佳 N/P 为 16∶1,CNE-miR-126 外观基本圆整、平均粒径(D50)为(144.79±1.85)nm、Zeta 电位为(22.92±1.24)mV;CNE-miR-126在1周内粒径及电位稳定性良好,且在48 h之内血清稳定性良好;在10-6～1 nniol/L浓度内对HUVECs细胞生长无明显抑制作用,能够促进HUVECs对miR-126的摄取;Weston blot结果显示,与Control组相比,LPS(1 mg/L)处理细胞8 h 后 VCAM-1蛋白水平显著升高(P＜0.01);与Model组相比,CNE-miR-126组预处理24 h后VCAM-1蛋白水平降低(P＜0.05).结论 CNE-miR-126粒径适宜,稳定性良好,可降低VCAM-1蛋白的表达,可为心血管疾病的治疗提供参考.
Objective To evaluate pharmaceutical properties,time stability,serum stability,cytotoxicity and cellular uptake of miR-126 cationic nanoemulsion(CNE)delivery system(CNE-miR126)loaded by CNE and assess the effect of CNE-miR126 on protein expression of vascular adhesion molecule-1(VCAM-1).Methods Optimal nitrogen-phosphorus ratio(N/P)of CNE and miR-126 was observed by agarose gel electrophoresis.Pharmaceutical performance was evaluated using nanoparticle size and Zeta potential measured by Zetasizer Nano ZS.The stability and potential of CNE-miR-126 particle size within 1 week under 4 ℃ were evaluated.Serum stability was used to assess the stability of CNE-miR-126 within 48 hours.MTT assay was applied to evaluate the cytotoxicity of CNE-miR-126.Flow cytometry and fluorescence microscopy were used to observe the uptake of CNE-miR-126 by human umbilical vein endothelial cells(HUVECs).Western blot was performed to detect the effect of CNE-miR-126 on VCAM-1 expression in inflammatory HUVECs induced by lipopolysaccharide(LPS).Results The optimal N/P ratio for CNE and miR-126 was 16:1.The appearance of CNE-miR-126 was basically round,with an average particle size(D50,144.79±1.85)nm and a Zeta potential(22.92±1.24)mV.CNE-miR-126 had good particle size and potential stability within 1 week,and serum stability was good within 48 hours.CNE-miR-126 at the concentration ranging from 10-6 to 1 nmol/L had no obvious inhibitory effect on HUVEC growth and promoted the uptake of miR-126 by HUVECs.Western blot result showed that VCAM-1 protein level was significantly upregulated after LPS(1 mg/L)treatment for 8 h when compared to control(P＜0.01).When compared to model group,CNE-miR-126 group showed a decrease in VCAM-1 protein level after 24 hours of pretreatment(P＜0.05).Conclusion CNE-miR-126 has suitable particle size and good stability,which can reduce VCAM-1 protein expression,providing a reference for cardiovascular disease therapy.

cationic nanoemulsionmiRNA-126gene deliverycellular uptakehuman umbilical vein endothelial cells

张丽丹、唐永瑜、汪巳卜、陈英、吴兴杰、沈祥春、陶玲

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贵州医科大学药学院,贵州省特色天然药物资源高效利用工程中心,贵州省高等学校天然药物药理与成药性评价特色重点实验室,天然药物资源优效利用重点实验室,贵州贵阳 550025

阳离子纳米乳 miRNA-126 基因递送 细胞摄取 人脐静脉内皮细胞

贵州省科技计划项目贵州省高层次创新型人才百层次人才项目贵州医科大学药学国际科技合作基地贵州省科技创新基地贵州省高层次创新型人才十层次人才

黔科合支撑[2020]4Y240贵州科技厅黔科合人才[2015]4029黔科合平台人才[2017]5802黔科合中引地[2023]003黔科合平台人才-GCC[2023]048

2024

10.19367/j.cnki.2096-8388.2024.06.004

贵州医科大学学报
贵阳医学院

贵州医科大学学报

CSTPCD
影响因子：0.827
ISSN：2096-8388
年,卷(期)：2024.49(6)
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