Preparation and cellular uptake study of anemoside B4 and PD-L1 siRNA co-delivered cRGD-modified targeting liposomes

WAN Anping; ZHANG Jing; ZHOU Xiong; FENG Yulin; LIU Jun; HE Yao; LI Xiang

doi:10.6039/j.issn.1001-0408.2023.01.04

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Preparation and cellular uptake study of anemoside B4 and PD-L1 siRNA co-delivered cRGD-modified targeting liposomes

更新时间：2023-02-22

- Preparation and cellular uptake study of anemoside B4 and PD-L1 siRNA co-delivered cRGD-modified targeting liposomes
- ZHONGGUO YAOFANG Vol. 34, Issue 1, Pages: 18-22(2023)
- 作者机构：
  
  1.江西中医药大学中药固体制剂制造技术国家工程研究中心，南昌　330006
  2.江西中医药大学现代中药制剂教育部重点实验室，南昌　330004
- 作者简介：
- 基金信息：
- DOI：10.6039/j.issn.1001-0408.2023.01.04
  CLC： R943
- Published：15 January 2023，
  
  Received：31 May 2022，
  
  Revised：05 December 2022，
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万安平,张婧,周雄等.白头翁皂苷B4和PD-L1 siRNA共递送cRGD修饰靶向脂质体的制备及体外细胞摄取研究 ^Δ[J].中国药房,2023,34(01):18-22.

WAN Anping,ZHANG Jing,ZHOU Xiong,et al.Preparation and cellular uptake study of anemoside B4 and PD-L1 siRNA co-delivered cRGD-modified targeting liposomes[J].ZHONGGUO YAOFANG,2023,34(01):18-22.
万安平,张婧,周雄等.白头翁皂苷B4和PD-L1 siRNA共递送cRGD修饰靶向脂质体的制备及体外细胞摄取研究 ^Δ[J].中国药房,2023,34(01):18-22. DOI： 10.6039/j.issn.1001-0408.2023.01.04.

WAN Anping,ZHANG Jing,ZHOU Xiong,et al.Preparation and cellular uptake study of anemoside B4 and PD-L1 siRNA co-delivered cRGD-modified targeting liposomes[J].ZHONGGUO YAOFANG,2023,34(01):18-22. DOI： 10.6039/j.issn.1001-0408.2023.01.04.

摘要

目的

制备白头翁皂苷B4（AB4）和程序性死亡配体1（PD-L1）小干扰RNA（siP）共递送环精氨酰甘氨酸天冬氨酸序列（cRGD）修饰靶向脂质体（AB4/siP-c-L），并考察其体外细胞摄取行为。

方法

采用乙醇注入法制备cRGD修饰的AB4靶向脂质体（AB4-c-L），将AB4-c-L与20 nmol/L的siP等体积混合，通过静电吸附得到AB4/siP-c-L。分别采用激光散射粒径测定仪、透射电子显微镜、超滤离心法、透析法、琼脂糖凝胶电泳法考察AB4/siP-c-L的粒径、Zeta 电位、形态、包封率、药物含量、体外释放行为、血清稳定性。分别采用流式细胞术和共聚焦激光扫描技术评价小鼠Lewis肺癌细胞LLC对AB4/siP-c-L的摄取及其在细胞内的分布情况。

结果

AB4/siP-c-L的平均粒径为（187.4±3.1） nm，Zeta电位为（33.5±1.4） mV，形态呈类球形，AB4的包封率和含量分别为（95.2±0.4）%、（1.0±0.2） mg/mL；AB4/siP-c-L可较好地包裹siP，并具有较好的血清稳定性、pH敏感性以及缓释特性。LLC细胞对AB4/siP-c-L的摄取率显著高于游离药物，并能实现细胞质内富集。

结论

AB4/siP-c-L可有效实现AB4与基因药物siP共载，且对LLC细胞具有一定的体外靶向性。

Abstract

OBJECTIVE

To prepare anemoside B4 （AB4） and programmed cell death ligand 1 （PDL1） siRNA （siP） co-delivered cRGD-modified targeting liposomes （AB4/siP-c-L）， and to study the cellular uptake

in vitro

METHODS

The cRGD-modified AB4-loaded targeted liposomes （AB4-c-L） were prepared by ethanol injection. AB4-c-L was mixed with 20 nmol/L siP in the same volume and AB4/siP-c-L was obtained through electrostatic adsorption. The particle size， Zeta potential， morphology， encapsulation efficiency and drug content，

in vitro

release behavior and serum stability of AB4/siP-c-L were investigated by laser scattering particle size tester， transmission electron microscopy， ultrafiltration centrifugation， dialysis and agar-gel electrophoresis block test. Cellular uptake of AB4/siP-c-L by Lewis lung cancer cells LLC and its intracellular localization were evaluated by flow cytometry and confocal laser scan technique.

RESULTS

The average particle size of AB4/siP-c-L was （187.4±3.1） nm， and the Zeta potential was （33.5±1.4） mV. AB4/siP-c-L was spheroidal in shape. The encapsulation efficiency and content of AB4 were （95.2±0.4） % and （1.0±0.2） mg/mL， respectively. AB4/siP-c-L could better package siP， and exhibited good serum stability， obvious pH sensitivity and sustained release property. The uptake rate of AB4/siP-c-L by LLC cells was significantly higher than that of free drug， and was able to accumulate in cytoplasm.

CONCLUSIONS

AB4/siP-c-L can effectively realize the co-loading of AB4 and gene drug siP， which has certain

in vitro

targeting to LLC cells.

关键词

白头翁皂苷B4程序性死亡配体1小干扰RNA脂质体细胞摄取

Keywords

PD-L1siRNAliposomecellular uptake

references

国家药典委员会. 中华人民共和国药典：一部[S]. 2020年版.北京：中国医药科技出版社，2020：108.

袁进，张丽峰，高红伟，等.白头翁皂苷B4药理作用研究进展[J].中成药，2022，44（7）：2229-2233.

狄孟华，李桂凤，李言，等. 白头翁皂苷B4在大鼠体内药代动力学规律及肺组织分布[J]. 贵州医科大学学报，2018，43（7）：745-749.

GHOSH C，LUONG G，SUN Y. A snapshot of the PD-1/PD-L1 pathway[J]. J Cancer，2021，12（9）：2735-2746.

GAVRILOV K，SALTZMAN W M. Therapeutic siRNA：principles，challenges，and strategies[J]. Yale J Biol Med，2012，85（2）：187-200.

MU X P，ZHANG M，WEI A H，et al. Doxorubicin and PD-L1 siRNA co-delivery with stem cell membrane-coated polydopamine nanoparticles for the targeted chemoimmunotherapy of PCa bone metastases[J]. Nanoscale，2021，13（19）：8998-9008.

DOWDY S F. Overcoming cellular barriers for RNA therapeutics[J]. Nature Biotechnology，2017，35（3）：222-229.

ALMEIDA B，NAG O K，ROGERS K E，et al. Recent progress in bioconjugation strategies for liposome-mediated drug delivery[J]. Molecules，2020，25（23）：5672.

NIEBERLER M，REUNING U，REICHART F，et al. Exploring the role of RGD-recognizing integrins in cancer[J]. Cancers，2017，9（9）：116.

JIN Y，TONG D Y，CHEN J N，et al. Overexpression of osteopontin，αvβ3 and Pim-1 associated with prognostically important clinicopathologic variables in non-small cell lung cancer[J]. PLoS One，2012，7（10）：e48575.

刘骏，罗莹，张婧，等. 白头翁皂苷B4长循环脂质体的处方优化与评价[J].中国新药杂志，2021，30（16）：1524-1529.

国家药典委员会. 中华人民共和国药典：四部[S]. 2020年版.北京：中国医药科技出版社，2020：61.

陈颖翀，李翔，张婧，等. 硫酸长春新碱脂质体含量测定及3种包封率测定方法的比较[J]. 中国药房，2014，25（37）：3504-3507.

LI X，ZOU Q，ZHANG J，et al. Self-assembled dual-targeted epirubicin-hybrid polydopamine nanoparticles for combined chemo-photothermal therapy of triple-negative breast cancer[J]. Int J Nanomedicine，2020，15：6791-6811.

O’MAHONY A M，DESGRANGES S，OGIER J，et al. In vitro investigations of the efficacy of cyclodextrin-siRNA complexes modified with lipid-PEG-Octaarginine：towards a formulation strategy for non-viral neuronal siRNA delivery[J]. Pharm Res，2013，30（4）：1086-1098.

ZHANG J，LI X，HUANG L. Anticancer activities of phytoconstituents and their liposomal targeting strategies against tumor cells and the microenvironment[J]. Adv Drug Deliv Rev，2020，（154/155）：245-273.

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