目的 制備葉酸（FA）修飾的pH值響應(yīng)型去甲斑蝥素（Nor）脂質(zhì)體（Nor@LP-CHS-FA），初步評價其促HepG2細(xì)胞凋亡活性。方法 應(yīng)用薄膜分散法制備Nor@LP-CHS-FA，采用單因素實驗結(jié)合星點設(shè)計-效應(yīng)面法設(shè)計和優(yōu)化制劑處方；研究該載藥系統(tǒng)的粒徑、ζ電位、多分散系數(shù)（PDI）、包封率、載藥量，應(yīng)用透射電鏡、傅里葉變換紅外光譜、差式掃描量熱儀、X射線儀進(jìn)行物理表征；評價其穩(wěn)定性及在模擬人工胃液、腸液和腫瘤微環(huán)境下的釋藥情況；通過溶血性實驗考察生物安全性，CCK-8法評估Nor@LP-CHS-FA對HepG2細(xì)胞的增殖抑制作用，結(jié)合流式細(xì)胞術(shù)分析其對HepG2細(xì)胞凋亡、周期的影響。結(jié)果 成功制備外觀呈黃色的Nor@LP-CHS-FA，其最優(yōu)處方為脂藥比5.05∶ 1、磷脂占膜材總質(zhì)量的52.85%、FA用量13.90 mg；透射電鏡下Nor@LP-CHS-FA呈規(guī)則球形，粒徑為（55.48±0.67） nm、ζ電位為（-18.15±0.54） mV、PDI為0.42±0.02、包封率（82.72±0.84）%、載藥量（12.25±0.13）%； Nor@LP-CHS-FA在4℃條件下儲存，包封率、載藥量更高，泄漏率更低。與游離Nor相比，Nor@LP-CHS-FA在模擬胃液、腸液和腫瘤微環(huán)境釋放速度更快，以模擬腫瘤微環(huán)境中最快，表明其有pH值響應(yīng)性，釋放行為遵循Weibull模型。溶血性實驗結(jié)果表明Nor@LP-CHS-FA有較小的溶血率，CCK-8結(jié)果顯示，給藥24、48、72 h后Nor@LP-CHS-FA對HepG2細(xì)胞的半數(shù)抑制濃度（IC50）分別為8.56、5.89、4.77 μg·mL^-1，流式細(xì)胞術(shù)結(jié)果表明Nor@LP-CHS-FA可誘導(dǎo)HepG2細(xì)胞發(fā)生凋亡和G2/M期阻滯。結(jié)論 Nor@LP-CHS-FA處方工藝簡單，具備良好的pH值響應(yīng)性，能增強藥物促HepG2細(xì)胞凋亡效果。

Objective To prepare pH-responsive norcantharidin (Nor) liposomes modified with folic acid (FA) (Nor@LP-CHS-FA) and preliminarily evaluate its activity in promoting apoptosis of HepG2 cells. Methods Nor@LP-CHS-FA was prepared by the thinfilm dispersion method. The formulation recipe was designed and optimized through single-factor experiments combined with central composite design-response surface methodology. The particle size, ζ potential, polydispersity index (PDI), encapsulation rate, and drug loading capacity were studied. Physical characterization examinations were conducted using transmission electron microscopy, fourier transform infrared spectroscopy, differential scanning calorimetry, and X-ray. The stability of the system was evaluated, as well as its drug release behavior in simulated artificial gastric fluid, intestinal fluid, and tumor microenvironment. The biological safety was evaluated through hemolysis experiments. The proliferation inhibitory effect of Nor@LP-CHS-FA on HepG2 cells was assessed using the CCK-8 method, and the effects on HepG2 cell apoptosis and cell cycle were analyzed by flow cytometry. Results The yellowcolored Nor@LP-CHS-FA was successfully prepared. The optimal formulation was a lipid-to-drug ratio of 5.05∶ 1, phospholipids accounting for 52.85% of the total mass of the membrane material, and a FA dosage of 13.90 mg. Under transmission electron microscopy, Nor@LP-CHS-FA presented a regular spherical shape, with a particle size of (55.48 ±0.67) nm, a ζ potential of (-18.15 ±0.54) mV, a PDI of 0.42 ±0.02, a encapsulation rate of (82.72 ±0.84)%, and a drug loading of (12.25 ±0.13)%. Nor@LP-CHS-FA had a higher encapsulation rate and drug loading and a lower leakage rate when stored at 4 ℃. Compared with free Nor, Nor@LP-CHS-FA released more rapidly in simulated gastric fluid, intestinal fluid, and tumor microenvironment, and was the fastest in the simulated tumor microenvironment, indicating its pH responsiveness and release behavior following the Weibull model. The hemolysis experiment results showed that Nor@LP-CHS-FA had a lower hemolysis rate. The CCK-8 results indicated that the half-maximal inhibitory concentration (IC₅₀) of Nor@LP-CHS-FA on HepG2 cells after 24, 48, and 72 hours of administration was 8.56, 5.89, 4.77 μg·mL^-1, respectively. The flow cytometry results showed that Nor@LP-CHS-FA could induce HepG2 cells to undergo apoptosis and G2/M phase arrest. Conclusion Nor@LP-CHS-FA prescription process is simple, has good pH responsiveness, and can enhance the effect of promoting HepG2 cell apoptosis by the drug.

R283.6

國家衛(wèi)生健康委醫(yī)院管理研究所項目（ YLZLXZ24G039）；四川省自然科學(xué)基金面上項目（2025ZNSFSC0684）；云南省科學(xué)技術(shù)廳地方高校聯(lián)合專項-面上項目（202001BA070001-041）；云南省天然藥物藥理重點實驗室開放基金重點項目（ YKLPNP-K2302)；四川省中醫(yī)藥管理局科研課題（ 2023MS057, 2023MS207）；四川省醫(yī)院協(xié)會青年藥師科研專項資金項目（ 22009, YP2202419）；攀枝花市科學(xué)技術(shù)局項目（2024ZD-S-28）