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
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篇名: | 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 |
TITLE: | Preparation of curcumin solid lipid nanoparticles inhalation powder loaded with nanoporous flower-shaped lactose and its in vitro inhibition effect on apoptosis |
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AnnexinⅤ/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,30s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5.809mg/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。 |
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 process of Cur-SLN-FL is simple and feasible. Cur-SLN-FL can improve LPS-induced apoptosis of BEAS-2B cells, and this effect is related to the regulation of mitochondrial membrane potential. |
期刊: | 2023年第34卷第02期 |
作者: | 李楠;李旭;程鹏;孔令钰;杨萍 |
AUTHORS: | LI Nan,LI Xu,CHENG Peng,KONG Lingyu,YANG Ping |
关键字: | 姜黄素;花形乳糖;固体脂质纳米粒;干粉吸入剂;凋亡 |
KEYWORDS: | curcumin; flower-shaped lactose; solid lipid |
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