Page 28 - 《中国药房》2023年2期

纳米多孔花形乳糖装载姜黄素固体脂质纳米粒吸入微粉的制备

及其体外抑凋亡作用研究
Δ

＊
李 楠 ，李 旭，程 鹏，孔令钰，杨 萍（天津市医药科学研究所，天津 300020）
#
中图分类号 R943；R965 文献标志码 A 文章编号 1001-0408（2023）02-0150-06
DOI 10.6039/j.issn.1001-0408.2023.02.05

摘 要 目的 制备纳米多孔花形乳糖（FL）装载姜黄素（Cur）固体脂质纳米粒（SLN）吸入微粉（Cur-SLN-FL），探讨其对脂多糖
（LPS）诱导的BEAS-2B细胞凋亡的抑制作用。方法 以不同种类（乳糖、蔗糖、甘露醇、海藻糖）和不同用量（2%、3%、5%）的冻干保
护剂为考察对象，采用冷冻干燥技术将 Cur-SLN 混悬液微粉化制成冻干粉，再将冻干粉与 FL 混匀后过 200 目筛，即得 Cur-SLN-
FL。采用扫描电镜、激光粒度仪等对 Cur-SLN-FL 的理化性质进行表征。以 BEAS-2B 细胞为对象，采用 Annexin Ⅴ/PI 双染法和
JC-1 试剂盒检测 LPS 诱导的 BEAS-2B 细胞经 Cur-SLN-FL 处理后的凋亡情况和线粒体膜电位变化。结果 以 3% 海藻糖为 Cur-
SLN冻干保护剂，所得冻干粉外形致密、饱满、不回缩塌陷，为色泽均匀的淡黄色粉末，30 s可完全溶解。当FL与Cur-SLN冻干粉
以质量比 1∶2 混匀时具有更高的二级分布沉积率[（40.92±0.02）%]。制得的 Cur-SLN-FL 呈花形外貌，平均粒径为（4.95±0.57）
μm，空气动力学粒径为（4.03±0.40）μm，临界相对湿度约为54%，排空率为（90.34±1.21）%，在2～7级收集盘中的可吸入细颗粒
量为（47.5±0.7）%，实测空气动力学粒径为（4.33±0.08）μm。Cur-SLN-FL 对 BEAS-2B 细胞半致死剂量（LD50 ）为 5.809 mg/mL。
Cur-SLN-FL 处理后的模型细胞凋亡率显著降低，线粒体膜电位显著升高（P＜0.05）。结论 Cur-SLN-FL 的制备工艺简单可行。
Cur-SLN-FL可改善LPS诱导的BEAS-2B细胞凋亡，且该作用与调节线粒体膜电位有关。
关键词 姜黄素；花形乳糖；固体脂质纳米粒；干粉吸入剂；凋亡

Preparation of curcumin solid lipid nanoparticles inhalation powder loaded with nanoporous flower-shaped
lactose and its in vitro inhibition effect on apoptosis
LI Nan，LI Xu，CHENG Peng，KONG Lingyu，YANG Ping（Tianjin Institute of Medical and Pharmaceutical
Science， Tianjin 300020， China）

ABSTRACT OBJECTIVE To prepare the nanoporous flower-shaped lactose （FL）-loaded curcumin （Cur） solid lipid
nanoparticles （SLN） inhalation powder （Cur-SLN-FL）， and to investigate its inhibition effect on LPS-induced apoptosis of BEAS-
2B cells. METHODS Using different kinds （lactose， sucrose， mannitol， trehalose） and different amounts （2%， 3%， 5%） of
freeze-dried protectants as objects， the suspension of Cur-SLN was micronized by freeze-drying technology into lyophilized
powder， which was then mixed with FL and sieved by a 200-mesh sieve to obtain Cur-SLN-FL. The physicochemical properties of
Cur-SLN-FL was characterized by scanning electron microscopy and laser particle size analyzer. Using BEAS-2B cells cultured in
vitro as objects， LPS-induced apoptosis and the changes of mitochondrial membrane potential after treatment of Cur-SLN-FL were
detected by Annexin Ⅴ/PI double staining method and JC-1 kit. RESULTS With 3% trehalose as Cur-SLN freeze-dried protective
agent， the freeze-dried powder obtained was compact and full in shape， did not shrink and collapse， and was uniform in color and
light-yellow powder， which could be completely dissolved in 30 s. When FL and Cur-SLN freeze-dried powder were mixed at a
ratio of 1∶2， it had a higher deposition rate of secondary distribution [（40.92±0.02）%]. SEM results showed that Cur-SLN-FL had
a flower-shaped appearance with an average particle size of （4.95±0.57） μm and an aerodynamic particle size of （4.03±0.40）
μm. The critical relative humidity of Cur-SLN-FL was about 54%， and the evacuation rate was （90.34 ± 1.21）%； the quantity of
fine particles that could be inhaled by Cur-SLN-FL in the 2-7 receiving discs was （47.5±0.7）%， and the measured aerodynamic
particle size was （4.33±0.08） μm. The LD50 of Cur-SLN-FL to BEAS-2B cells was 5.809 mg/mL. The apoptosis rate of model
cells was significantly reduced after treatment of Cur-SLN-FL， and the mitochondrial membrane potential was significantly
increased （P＜0.05）. CONCLUSIONS The preparation
Δ 基金项目 国家自然科学基金资助项目（No.81903565）；天津市 process of Cur-SLN-FL is simple and feasible. Cur-SLN-FL
科技计划项目（No.21JCYBJC01630）；天津市卫生健康科技项目（No.
can improve LPS-induced apoptosis of BEAS-2B cells， and
TJWJ2021MS046，No.TJWJ2021QN078）
this effect is related to the regulation of mitochondrial
＊第一作者 助理研究员，硕士。研究方向：纳米给药系统。
E-mail：llittlsnows@126.com membrane potential.
# 通信作者 副 研 究 员 。 研 究 方 向 ：药 物 制 剂 。 E-mail： KEYWORDS curcumin； flower-shaped lactose； solid lipid
tjyangping74@163.com nanoparticles； dry powder inhalers； apoptosis

· 150 · China Pharmacy 2023 Vol. 34 No. 2 中国药房 2023年第34卷第2期