目的 構(gòu)建pH敏感/葉酸修飾白藜蘆醇脂質(zhì)體(pH-sensitive/folic acid-modified resveratrol liposomes,pH/FA-Res-Lips),并進行理化性質(zhì)和抗腫瘤評價。方法 單因素法考察pH/FA-Res-Lips主要影響因素,使用Box-Behnken設(shè)計-效應(yīng)面法(Box-Behnken design-response surface method,BBD-RSM)優(yōu)化pH/FA-Res-Lips處方工藝。采用紫外-可見光吸收光譜(ultraviolet-visible absorption spectroscopy,UV)、傅里葉變換紅外光譜法(Fourier transform infrared spectroscopy,FT-IR)、透射電子顯微鏡(transmission electron microscope,TEM)進行表征,考察pH/FA-Res-Lips在不同pH值磷酸鹽緩沖液(PBS)中的釋藥行為。采用MTT法和Annexin V/PI雙染法考察pH/FA-Res-Lips對人子宮內(nèi)膜癌Ishikawa細胞的生長抑制及促凋亡作用。采用Ishikawa細胞建立子宮內(nèi)膜癌荷瘤小鼠模型,分為模型對照組(生理鹽水)、陽性對照組(順鉑,2.0 mg/kg)、白藜蘆醇組(15 mg/kg)、白藜蘆醇脂質(zhì)體組(Res-Lips,15 mg/kg)、pH-Res-Lips組(15 mg/kg)、FA-Res-Lips組(15 mg/kg)、pH/FA-Res-Lips低劑量組(10 mg/kg)和pH/FA-Res-Lips高劑量組(15 mg/kg),比較pH/FA-Res-Lips體內(nèi)抗腫瘤效果。通過血液生化指標(biāo)和小鼠主要臟器HE染色來評價pH/FA-Res-Lips安全性。結(jié)果 pH/FA-Res-Lips最佳處方為磷脂與白藜蘆醇用量比為5.80∶1,磷脂與膽固醇用量比為5.00∶1,磷脂與DSPE-PEOz-FA用量比為5.30∶1。pH/FA-Res-Lips平均包封率、載藥量、平均粒徑和ζ電位分別為(85.74±1.09)%、(8.77±0.11)%、(192.25±5.38)nm和(-28.12±1.20)mV。pH/FA-Res-Lips未影響白藜蘆醇化學(xué)結(jié)構(gòu),白藜蘆醇與脂質(zhì)材料之間存在氫鍵絡(luò)合作用。pH/FA-Res-Lips外貌呈囊泡狀。pH/FA-Res-Lips在pH 5.5、6.0 PBS中累積釋放度較高,具有明顯的pH敏感性。MTT法結(jié)果顯示,白藜蘆醇和pH/FA-Res-Lips對Ishikawa細胞半數(shù)抑制濃度(IC₅₀)分別為98.14、48.43 μg/mL,并有效誘導(dǎo)細胞凋亡。pH/FA-Res-Lips顯著抑制了子宮內(nèi)膜癌荷瘤小鼠腫瘤生長,體質(zhì)量未見顯著性下降,pH/FA-Res-Lips(15 mg/kg)將白藜蘆醇(15 mg/kg)抑瘤率由17.35%提高至66.67%。pH/FA-Res-Lips未對荷瘤小鼠各個臟器產(chǎn)生影響,使用安全性較高。結(jié)論 pH/FA-Res-Lips顯著性提高了白藜蘆醇抗腫瘤作用,為進一步開發(fā)成抗腫瘤藥物提供了實驗依據(jù)。

Objective The pH-sensitive/folic acid-modified resveratrol liposomes (pH/FA-Res-Lips) was prepared, its physicochemical properties and antitumor effects were also studied. Methods The main influencing factors of pH/FA-Res-Lips were investigated by single factor experiments, Box-Behnken response-surface design method (BBD-RSM) was employed to optimize prescriptions of pH/FA-Res-Lips. pH/FA-Res-Lips was characterized by ultraviolet-visible absorption spectroscopy (UV), Fourier transform infrared spectroscopy (FT-IR) and transmission electron microscopy (TEM). Drug release behavior of pH/FA-Res-Lips in phosphate buffer saline solution (PBS) with different pH values was investigated. MTT method and Annexin V/PI double staining method were used to investigate the inhibitory and proapoptotic effects of pH/FA-Res-Lips on Ishikawa cells, respectively. Ishikawa cells were used to establish the mouse model of endometrial cancer, and divided into model control group (normal saline), positive control group (cisplatin, 2.0 mg/kg), resveratrol group (15 mg/kg), resveratrol liposomes (Res-Lips, 15 mg/kg) group, pH-sensitive resveratrol liposomes (pH-Res-Lips, 15 mg/kg) group, FA-modified resveratrol liposomes (FA-Res-Lips, 15 mg/kg) group, low-dose pH/FA-Res-Lips (10 mg/kg) group and high-dose pH/FA-Res-Lips (15 mg/kg) group, the in vivo antitumor effects of pH/FA-Res-Lips were compared. Safety evaluation of pH/FA-Res-Lips was performed by blood biochemical indexes and HE staining of the main organs of mice. Results Optimal formulation of pH/FA-Res-Lips nanoparticles as follows: phospholipids to resveratrol ratio was 5.80:1, phospholipids to cholesterol ratio was 5.00:1, and phospholipids to DSPE-PEOz-FA ratio was 5.30:1. Average envelopment efficiency, drug loading, particle size and ζ potential were (85.74 ± 1.13)%, (8.77 ± 0.16)%, (192.25 ± 7.61) nm and (-28.12 ± 1.20) mV, respectively. pH/FA-Res-Lips did not affect the structure of resveratrol, and there was a hydrogen bond complexation between resveratrol and lipid materials. The appearance of pH/FA-Res-Lips was vesicular. pH/FA-Res-Lips existed higher cumulative release in PBS of pH 5.5 and 6.0, which demonstrated that pH/FA-Res-Lips had obvious pH-sensitivity. MTT method results showed that IC50 of resveratrol and pH/FA-Res-Lips on Ishikawa cells were 98.14, 48.43 μg/mL, respectively, and effectively induced apoptosis of Ishikawa cells. pH/FA-Res-Lips significantly inhibited the tumor growth of endometrial cancer-bearing mice, and there was no significant decrease in body weight. pH/FA-Res-Lips (15 mg/kg) increased the tumor inhibition rate of resveratrol (15 mg/kg) from 17.35% to 66.67%. pH/FA-Res-Lips did not affect the organs of tumor-bearing mice, which showed that the safety was relatively high. Conclusion pH/FA-Res-Lips significantly enhanced the anti-tumor efficacy of resveratrol, which had the potential to be developed into an antitumor drug.

R283.6

2025年度河南省科技攻關(guān)項目（252102310468）；河南省高等學(xué)校重點科研項目計劃（23B320013）；河南應(yīng)用技術(shù)職業(yè)學(xué)院2023年度校級課題（2023-K.J-54）；河南省醫(yī)學(xué)科技攻關(guān)計劃（SBGJ202102153）