目的 制備雷公藤紅素納米混懸劑，并進(jìn)行體內(nèi)外抗腫瘤作用研究。方法 采用超聲注入聯(lián)合旋轉(zhuǎn)蒸發(fā)法制備雷公藤紅素納米混懸劑，以粒徑大小為指標(biāo)，篩選合適的穩(wěn)定劑；采用動態(tài)光散射法、透射電鏡考察粒徑和形態(tài)，并對其不同介質(zhì)穩(wěn)定性、體外釋放、溶血性、體外抗腫瘤活性進(jìn)行研究；建立H22荷瘤小鼠模型，以雷公藤紅素注射液為對照組，2 mg/kg iv給藥，考察體內(nèi)抗腫瘤作用。結(jié)果 聚乙二醇-聚己內(nèi)酯（mPEG2000-PCL2000）為雷公藤納米混懸劑的優(yōu)良的穩(wěn)定劑，所制備的納米混懸劑粒徑為（67.1±3.0）nm，Zeta電位為（-10.4±1.45）mV，多分散性指數(shù)為0.232±0.08，近乎為球形，分布比較均勻。在磷酸緩沖液（PBS）、血漿、生理鹽水、5%葡萄糖中均穩(wěn)定；體外緩慢釋放，在144 h累積釋放率達(dá)到74.04%；MTT結(jié)果顯示雷公藤紅素納米混懸劑對HepG2細(xì)胞的毒性強(qiáng)于溶液（IC₅₀，1.179 μg/mL vs 2.377 μg/mL，P<0.05）。體內(nèi)研究中雷公藤紅素納米混懸劑對H22荷瘤小鼠的的抑瘤率顯著高于注射液組（70.36% vs 51.1%，P<0.05）。結(jié)論 制備的雷公藤紅素納米混懸劑粒徑小、載藥量高、穩(wěn)定性好，顯著提高了雷公藤紅素的抗腫瘤效果，可以作為雷公藤紅素作為抗腫瘤藥物應(yīng)用的合適劑型。

Objective To prepare Celastrol Nanoparticles and study its anti-tumor activities in vitro and in vivo. Methods Celastrol nanosuspensions were prepared by microprecipitation combined with evaporation method. Suitable stabilizer was screened according to the size of the resultant nanosuspensions. Dynamic light scattering method was used to measure the particle size and transmission electron microscopy was used to observe the morphology. The stability in different medium, drug release in vitro, the hemolysis and the cytotoxicity were studied. Anti-tumor effect in vivo was investigated on H22-bearing mice using Celastrol Injections as the control at the dose of 2 mg/kg. Results Polyethylene glycol-polycaprolactone (mPEG2000-PCL2000) was a good stabilizer for Celastrol Nanoparticles. Celastrol Nanoparticles had (67.1 ±3.0) nm in diameter with a polydisperse index of (0.23 ±0.08) and a Zeta potential of (-10.4 ±1.5) mV. Celastrol Nanoparticles were nearly spherical in morphology and had a more uniform distribution. They were quite stable in PBS, normal saline, 5% glucose, and plasma. Celastrol Nanoparticles showed sustained drug release and the cumulative release reached 74.04% within 144 h. MTT assay displayed that Celastrol Nanoparticles had stronger cytotoxicity against HepG2 cells than free c elastrol (IC₅₀, 1.179 μg/mL vs 2.377 μg/mL, P<0.05). It was demonstrated that Celastrol Nanoparticles significantly improved the therapeutic efficacy in vivo on H22-bearing mice in contrast to Celastrol Injections (70.36% vs 51.1%, P<0.05). Conclusion Celastrol nanosuspensions have characteristics of small particle size, high drug-loading content, and good stability, which significantly improve the anti-tumor effect of celastrol, and it is believed to be suitable dosage form for the application of celastrol in tumor treatment.

國家自然科學(xué)基金資助項(xiàng)目（U1401223，81460734）