越橘叶黄酮脂质体制备及对HepG2细胞的影响
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东北林业大学 生命科学学院

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TQ630????

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Preparation of flavonoid liposomes from lingonberry leaves and their effect on HepG2 cells
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College of Life Sciences,Northeast Forestry University

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    摘要:

    以越橘叶为主要原料,液质联用技术分析越橘叶主要成分,制备pH敏感型越橘叶黄酮脂质体,探究其在不同pH下体外释放以及对HepG2细胞抑制能力。采用超声辅助提取和大孔树脂纯化获得18组馏分,依据活性跟踪法分析最佳组分进行液质分析;采用乙醇注入法结合羧甲基壳聚糖表面修饰制备pH敏感型脂质体,通过傅里叶红外、粒径、多分散指数、?-电位和透析法对脂质体进行结构表征,分析脂质体体外释放能力;通过增殖能力分析、迁移能力分析和对细胞周期的影响判断脂质体对HepG2细胞抑制作用。结果显示,大孔树脂可有效提高黄酮的纯度和抗氧化能力,纯化后黄酮浓度可达899.44 mg/g,HPLC-MS分析共得到9种成分。用此黄酮制备的脂质体包封率为81.77?3.02%,粒径为408.30?2.23 nm且粒径分布均匀,当羧甲基壳聚糖浓度为0.4%时,?-电位绝对值最大溶液稳定性最好。体外释放实验证明其具有pH敏感性且符合Weibull模型,细胞实验进一步证明了与未包裹羧甲基壳聚糖的脂质体相比其具有pH敏感性,脂质体增强了对HepG2细胞抑制能力并阻滞HepG2细胞停留在S期和G2期。

    Abstract:

    The main ingredient of the study is the leaves of the lingonberry. The liquid-liquid extraction technique was employed to analyze the primary constituents of Vietnamese kumquat leaves and to formulate pH-sensitive liposomes containing Vietnamese kumquat leaf flavonoids. The study also investigated their in vitro release at various pH levels and their inhibitory effect on HepG2 cells. Eighteen fractions were obtained using ultrasound-assisted extraction and macroporous resin purification. The optimal components were analyzed using the activity tracking method for liquid-liquid analysis. pH-sensitive liposomes were prepared using the ethanol injection method, which was combined with carboxymethyl chitosan surface modification. The liposomes were characterized using Fourier transform infrared spectroscopy, particle size analysis, polydispersity index measurement, zeta potential determination, and the dialysis method to analyze their in vitro release capability. The inhibitory effect of the liposomes on HepG2 cells was determined through analysis of cell proliferation, migration, and the cell cycle. The results showed that the macroporous resin effectively increased the purity and antioxidant capacity of the flavonoids. The concentration of purified flavonoids reached 899.44 mg/g. HPLC-MS analysis identified nine components, including epicatechin-3-O-cinnamate, catechin, dihydromyricetin, glycyrrhizic chalcone A, quercetin-3β-D-glucoside, kaempferol-3-O-L-rhamnoside, isorhamnetin-3-6-gallic acid glucoside, kaempferol 3-(6-gallic acid lactose), and quercetin 3-(2-gallic acid ellagic acid glucoside). The encapsulation efficiency of the liposomes prepared with this flavonoid is 81.77±3.02%, with a particle size of 408.30±2.23 nm and a uniform particle size distribution. When the concentration of carboxymethyl chitosan is 0.4%, the solution exhibits the highest stability, as indicated by the highest absolute value of x-potential. In vitro release experiments have shown its pH sensitivity, with a cumulative release of 98.03±3.18% in 48 hours at a release medium pH of 4.0. Additionally, it conforms to the Weibull model. Cell experiments further demonstrate that compared to liposomes without carboxymethyl chitosan encapsulation, liposomes with carboxymethyl chitosan encapsulation exhibit pH sensitivity, enhance the inhibitory ability of HepG2 cells, and block the HepG2 cells from staying in the S and G2 phases.

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成圆,丁淼,王思宇,樊梓鸾.越橘叶黄酮脂质体制备及对HepG2细胞的影响[J].精细化工,2024,41(9):

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  • 收稿日期:2023-10-16
  • 最后修改日期:2024-01-03
  • 录用日期:2023-11-21
  • 在线发布日期: 2024-09-10
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