Sustained-release genistein from nanostructured lipid carrier suppresses human lens epithelial cell growth

AIM： To design and investigate the efficacy of a modified nanostructured lipid carrier loaded with genistein（Gen-NLC） to inhibit human lens epithelial cells（HLECs） proliferation.·METHODS： Gen-NLC was made by melt emulsification method. The morphology, particle size（PS）, zeta potentials（ZP）, encapsulation efficiency（EE） and in vitro release were characterized. The inhibition effect of nanostructured lipid carrier（NLC）, genistein（Gen） and Gen-NLC on HLECs proliferation was evaluated by cell counting kit-8（CCK-8） assay, gene and protein expression of the proliferation marker Ki67 were evaluated with real-time quantitative polymerase chain reaction（RT-q PCR） and immunofluorescence analyses.·RESULTS： The mean PS of Gen-NLC was 80.12±1.55 nm with a mean polydispersity index of 0.11±0.02. The mean ZP was-7.14 ±0.38 m V and the EE of Gen in the nanoparticles was 92.3% ±0.73%. Transmission electron microscopy showed that Gen-NLC displayed spherical-shaped particles covered by an outer-layer structure. In vitro release experiments demonstrated a prolonged drug release for 72 h. The CCK-8 assay results showed the NLC had no inhibitory effect on HLECs and Gen-NLC displayed a much more prominent inhibitory effect on cellular growth compared to Gen of the same concentration. The m RNA and protein expression of Ki67 in LECs decreased significantly in Gen-NLC group.·CONCLUSION： Sustained drug release by Gen-NLCs may impede HLEC growth.

Capillary electrophoresis to determine entrapment efficiency of a nanostructured lipid carrier loaded with piroxicam

A simple and fast capillary electrophoresis method has been developed to determine the amount of piroxicam loaded in a drug delivery system based on nanostructured lipid carriers（NLCs）.The entrapment efficiency of the nanostructured lipid carrier was estimated by measuring the concentration of drug not entrapped in a suspension of NLC.The influence of different parameters on migration times,peak symmetry,efficiency and resolution was studied;these parameters included the pH of the electrophoretic buffer solution and the applied voltage.The piroxicam peak was obtained with a satisfactory resolution.The separation was carried out using a running buffer composed of 50 mM ammonium acetate and 13.75 mM ammonia at pH 9.The optimal voltage was 20 kV and the cartridge temperature was 20 ℃.The corresponding calibration curve was linear over the range of 2.7-5.4 μg/mL of NLC suspension.The reproducibility of migration time and peak area were investigated,and the obtained RSD%values（n = 5） were 0.99 and 2.13.respectively.

A hybrid genipin-crosslinked dual-sensitive hydrogel/nanostructured lipid carrier ocular drug delivery platform

The objective of this study was to develop a novel hybrid genipin-crosslinked dual-sensitive hydrogel/nanostructured lipid carrier(NLC) drug delivery platform. An ophthalmic antiinflammatory drug, baicalin(BN) was chosen as the model drug. BN –NLC was prepared using melt-emulsification combined with ultra-sonication technique. Additionally, a dual pH-and thermo-sensitive hydrogel composed of carboxymethyl chitosan(CMCS) and poloxamer 407(F127) was fabricated by a cross-linking reaction with a nontoxic crosslinker genipin(GP). GP-CMCS/F127 hydrogel was characterized by FTIR, NMR, XRD and SEM. The swelling studies showed GP-CMCS/F127 hydrogel was both pH-and thermo-sensitive. The results of in vitro release suggested BN –NLC gel can prolong the release of baicalin comparing with BN eye drops and BN –NLC. Ex vivo cornea permeation study was evaluated using Franz diffusion cells. The apparent permeability coefficient(Papp) of BN –NLC gel was much higher(4.46-fold) than that of BN eye drops. Through the determination of corneal hydration levels, BN –NLC gel was confirmed that had no significant irritation to cornea. Ex vivo precorneal retention experiments were carried out by a flow-through approach. The results indicated that the NLC-based hydrogel can prolong precorneal residence time. In conclusion, the hybrid NLCbased hydrogel has a promising potential for application in ocular drug delivery.

Solid Lipid Nanoparticles (SLN) and Nanostructured Lipid Carriers (NLC): Occlusive Effect and Penetration Enhancement Ability

Objective: This work compares the occlusive effect and the penetration enhancement ability of solid lipid nanoparticles (SLN) and nanostructured lipid carriers (NLC), through in vitro skin. Methods: SLN and NLC were prepared by high shear homogenization and characterized by size, polydispersity index, zeta potential, morphology and physical stability. Occlusive effect was assessed by an in vitro test and by measuring TEWL using pig skin. Skin treated with the lipid carriers was visualized by SEM. A penetration test through skin, followed by tape stripping, was carried out using Nile red as a marker. Results: SLN (200 ± 6 nm) and NLC (192 ± 11 nm) were obtained. An occlusion factor of 36% - 39% was observed for both systems, while a reduction in TEWL of 34.3% ± 14.8% and 26.2% ± 6.5% was seen after treatment with SLN and NLC, respectively. SEM images showed a film formed by the lipid carriers, responsible for the occlusion observed. No differences were found between the occlusive effect produced by SLN and NLC in both tests. NLC allowed the penetration of a greater amount of Nile red than SLN: 4.7 ± 1.3 μg and 1.7 ± 0.4 μg, respectively. Conclusion: Both carriers form a film on the skin, providing an occlusive effect with no differences between these two systems. The penetration of a marker (Nile red) into the stratum corneum was quite higher for NLC than for SLN, suggesting an influence of the composition of these particles on their penetration enhancing ability.

Preparation and Characterization of Nanostructured Lipid Carrier Loaded with UVA/UVB Filters

A new photoprotective system based on encapsulating UVA (butyl methoxydibenzoylmethane, BMBM) and UVB (octyl methoxycinnamate, OMC) filters into nanostructured lipid carriers (NLC) has been prepared to develop cosmetic formulations with effective UV protection. BMBM/OMC-loaded NLC was prepared by ultrasonication-homogenisation, and analysed by particle size, zeta potential (ZP), encapsulation efficiency (EE), scanning electron microscopy (SEM), fourier transform infrared spectroscopy (FTIR) and differential scanning calorimetry (DSC). Moreover, the UV protection property and photostability were investigated and compared with BMBM/OMC-conventional emulsions. The particle size and ZP of BMBM/OMC-loaded NLC were 310.24 nm and -33.6 mV, EE of BMBM and OMC were 85.46% and 99.32%. SEM, FTIR and DSC analysis confirmed BMBM and OMC entrapped in the lipid matrix core and the structure was stable during storage. Compared with conventional emulsion, BMBM/OMC-loaded NLC displayed perfect photo protection property in whole UV range. The photostability studies showed that the NLC can improve the photostability of sunscreens.

Lipid carriers for mRNA delivery

Messenger RNA(mRNA)is the template for protein biosynthesis and is emerging as an essential active molecule to combat various diseases,including viral infection and cancer.Especially,mRNA-based vaccines,as a new type of vaccine,have played a leading role in fighting against the current global pandemic of COVID-19.However,the inherent drawbacks,including large size,negative charge,and instability,hinder its use as a therapeutic agent.Lipid carriers are distinguishable and promising vehicles for mRNA delivery,owning the capacity to encapsulate and deliver negatively charged drugs to the targeted tissues and release cargoes at the desired time.Here,we first summarized the structure and properties of different lipid carriers,such as liposomes,liposome-like nanoparticles,solid lipid nanoparticles,lipid-polymer hybrid nanoparticles,nanoemulsions,exosomes and lipoprotein particles,and their applications in delivering mRNA.Then,the development of lipid-based formulations as vaccine delivery systems was discussed and highlighted.Recent advancements in the mRNA vaccine of COVID-19 were emphasized.Finally,we described our future vision and perspectives in this field.

Nanostructured lipid carrier based topical gel of Ganoderma Triterpenoids for frostbite treatment

The objective of this study was to prepare nanostructured lipid carrier(NLC)-based topical gel of Ganoderma Triterpenoids(GTs) and evaluate their effects on frostbite treatment. GT-NLCs was prepared by the high pressure homogenization method and then characterized by morphology and analyses of particle size, zeta potential, entrapment efficiency(EE), and drug loading(DL). The NLCs was suitably gelled for skin permeation studies in vitro and pharmacodynamic evaluation in vivo, compared with the GT emulgel. The GT-NLC remained within the colloidal range and was uniformly dispersed after suitably gelled by carbopol preparation. Transmission electron microscopy(TEM) study showed GT-NLCs was spherical in shape. The EE(%) and DL(%) could reach up to(81.84 ± 0.60)% and(2.13 ± 0.12)%, respectively. The result of X-ray diffractograms(XRD) showed that GTs were in an amorphous state in the NLC-gel. In vitro permeation studies through rat skin indicated that the amount of GTs permeated through skin of GT-NLCs after 24 h was higher than that of GT emulsion, and GT-NLCs increased the accumulative amounts of GTs in epidermis 7.76 times greater than GT emulsion. GT-NLC-gel was found to possess superior therapeutic effect for frostbite, compared with the GT emulgel. The NLC based topical gel of GTs could improve-their therapeutic effect for frostbite.

Intranasal delivery of nanostructured lipid carriers,solid lipid nanoparticles and nanoemulsions:A current overview of in vivo studies

The management of the central nervous system(CNS)disorders is challenging,due to the need of drugs to cross the blood-brain barrier(BBB)and reach the brain.Among the various strategies that have been studied to circumvent this challenge,the use of the intranasal route to transport drugs from the nose directly to the brain has been showing promising results.In addition,the encapsulation of the drugs in lipid-based nanocarriers,such as solid lipid nanoparticles(SLNs),nanostructured lipid carriers(NLCs)or nanoemulsions(NEs),can improve nose-to-brain transport by increasing the bioavailability and site-specifc delivery.This review provides the state-of-the-art of in vivo studies with lipid-based nanocarriers(SLNs,NLCs and NEs)for nose-to-brain delivery.Based on the literature available from the past two years,we present an insight into the different mechanisms that drugs can follow to reach the brain after intranasal administration.The results of pharmacokinetic and pharmacodynamics studies are reported and a critical analysis of the differences between the anatomy of the nasal cavity of the different animal species used in in vivo studies is carried out.Although the exact mechanism of drug transport from the nose to the brain is not fully understood and its effectiveness in humans is unclear,it appears that the intranasal route together with the use of NLCs,SLNs or NEs is advantageous for targeting drugs to the brain.These systems have been shown to be more effective for nose-to-brain delivery than other routes or formulations with non-encapsulated drugs,so they are expected to be approved by regulatory authorities in the coming years.

基于耗散粒子动力学模拟辅助的纳米结构脂质载体优化制备

基于分子模拟能够探究纳米结构脂质载体(nanostructure lipid carriers,NLC)各物质间的结合能、溶解度参数等,预测混合物的相容性,从而用于指导实验前原料种类以及用量选择。该文以姜黄素(curcumin,Cur)为模型营养物,以固体脂质(单硬脂酸甘油酯)和液体脂质(中链甘油三酯、葵花籽油、甜橙油)及表面活性剂(泊洛沙姆188、卵磷脂、吐温-80)作为筛选对象,采用耗散粒子动力学(dissipative particle dynamics,DPD)方法进行模拟筛选,再通过正交试验制备NLC以对模拟结果进行验证,最后对NLC的粒径和微观形态等进行表征。模拟与实验结果均表明,单硬脂酸甘油酯-中链甘油三酯(medium chain triglycerides,MCT)-卵磷脂能形成较好的载体体系,固液脂质比为1∶3(质量比),卵磷脂含量为5%(质量分数),姜黄素含量占总脂质的3%(质量分数)时形成的球壳结构最好。优化后制备的姜黄素NLC平均粒径为(233.8±2.4)nm,电位为(-38.2±0.6)mV,包封率为85.06%,微观形态均匀分散、没有明显聚集现象。

紫草素纳米结构脂质载体的制备与评价

目的 制备并优化紫草素纳米结构脂质载体(NLC),并对其进行表征、创面刺激性测试及抗炎活性评价。方法 通过饱和溶解度测定、脂质相容性和乳化能力筛选了紫草素NLC的处方。以粒径和多分散系数(PDI)为指标,考察紫草素NLC的制备方法,并采用该方法制备紫草素NLC,单因素法考察固液脂质比、总脂质用量、乳化剂用量和投药量的范围。通过正交试验考察固液脂质比、总脂质用量、投药量对PDI和载药量的影响,得到最优处方,并对其进行验证及表征。通过在大鼠背部表皮造成破损后连续给药,测试紫草素NLC的创面刺激性。采用脂多糖诱导RAW264.7细胞炎症模型,以NO释放率为指标评价紫草素NLC的体外抗炎活性。结果 采用Transcutol P为液体脂质,固体脂质为双硬脂酸甘油酯,乳化剂为BRIJ?O20,采用薄膜水化法制备NLC,最优处方:乳化剂用量为6%,固液脂质比为4∶6,总脂质用量为6%,投药量为20 mg。大鼠背部破损皮肤均未见红斑或水肿,皮肤组织结构均基本正常,同时等剂量下的紫草素NLC抑制RAW264.7释放NO的能力存在比紫草素更强的趋势。结论 制得的紫草素NLC载药量高,粒径均一。紫草素被制成NLC后,对创面无刺激性,细胞毒性有所下降,同时具有较好的抗炎活性。

白藜芦醇纳米结构脂质载体的制备与体外释放研究

目的 制备白藜芦醇纳米结构脂质载体(Res-NLCs),并对其进行理化性质和体外释放特性的考察。方法 采用乳化蒸发-高压均质法制备Res-NLCs,通过正交试验得出最优处方,并对最优处方制得的Res-NLCs进行理化性质考察;通过HPLC建立白藜芦醇体外释放方法学,采用动态透析法对Res-NLCs的体外释药特性进行考察。结果 Res-NLCs的最优处方为:双硬脂酸甘油酯用量为0.0356 g,大豆磷脂用量为0.3918 g,Tween-80的质量浓度为4.25%,辛酸癸酸聚乙二醇甘油酯用量为0.0475 g。经透射电镜观察Res-NLCs呈规则圆球状,分散均匀。测得Res-NLCs的平均粒径为(81.87±0.14)nm,多分散系数为(0.099±0.02),Zeta电位为(-8.09±0.75)m V,包封率为(98.43±0.07)%,载药量为(11.04±0.14)%。体外释放结果显示,白藜芦醇溶液在8 h释放97.31%,而Res-NLCs则缓慢持续释放,并在48 h后达到释放平台,累计释放的白藜芦醇为80.69%。结论 通过乳化蒸发-高压均质法制备的Res-NLCs粒径小,分散均匀,具有较高的包封率及载药量,并可显著延长白藜芦醇释放的时间,具有良好的缓释效果。

脂质纳米粒在经皮给药系统中治疗皮肤病的应用

经皮给药作为一种非侵入性给药方式,正逐渐受到广大医药工作者和患者的青睐。然而,皮肤角质层的屏障功能限制了药物的经皮吸收。脂质纳米粒作为一种新型的药物载体能够有效提高皮肤渗透性、减少药物相关副作用。本文就脂质纳米粒的特点、透皮作用机制以及在多种皮肤病中的应用做一综述,阐明脂质纳米粒在经皮给药领域的广阔发展前景,为脂质纳米粒的开发及临床应用提供参考。

血浆NGAL、尿NAG及尿RBP在诊断早期糖尿病肾脏疾病中的价值

目的探讨血浆中性粒细胞明胶酶相关脂质运载蛋白(NGAL)、尿N-乙酰-β-D-氨基葡萄糖苷酶(NAG)和尿视黄醇结合蛋白(RBP)在诊断早期糖尿病肾脏疾病(DKD)中的价值。方法选取2021年5月至2023年3月在上海市浦东新区公利医院内分泌科确诊的2型糖尿病(T2DM)患者227例作为研究对象,根据尿微量清蛋白/肌酐比值(UACR)进一步分为正常蛋白尿(NA)组、微量蛋白尿(MA)组、大量蛋白尿(HA)组,选取88例体检健康者作为对照组。检测各组UACR、NGAL、NAG、RBP、血糖、肾功能水平。分析NGAL、NAG、RBP水平与DKD的相关性,采用受试者工作特征(ROC)曲线分析各项指标诊断早期DKD的价值。结果根据UACR水平分组,NA组97例,MA组73例,HA组57例。NA组、MA组和HA组的NGAL、NAG、RBP水平高于对照组,并且HA组>MA组>NA组,差异均有统计学意义(P<0.05)。Pearson相关分析发现,NGAL、NAG、RBP水平与UACR、尿素氮(BUN)、血肌酐(SCr)、光抑素C(CysC)均呈正相关(P<0.05)。ROC曲线分析结果显示,NGAL、NAG、RBP及三者联合检测诊断DKD的曲线下面积(AUC)依次为0.854、0.788、0.830、0.929;单项检测时NGAL的灵敏度(82.3%)较高,NGAL、NAG和RBP联合检测时诊断效能最大(AUC=0.929)。结论血浆NGAL、尿NAG、尿RBP水平与DKD患者肾脏损伤程度密切相关。三者联合检测可发挥其最大诊断效能,为早期DKD的诊断提供新的方法。

信使核糖核酸药物递送载体的研究进展

信使核糖核酸(mRNA)作为一类新兴的核酸类药物,在疫苗、免疫治疗、再生医学、基因编辑等方面具有巨大的应用潜力,尤其是2020年全球新型冠状病毒感染疫情极大加速mRNA技术的发展。然而,mRNA带负电,很难进入细胞,且在体内易被先天免疫系统的细胞吞噬或被核酸酶降解,其不稳定性和低递送效率阻碍了广泛应用。因此,需要安全、有效、稳定的递送载体,以保护mRNA不被降解,突破细胞膜障碍,实现细胞内表达。该文系统介绍了基于mRNA药物递送载体的最新研究进展,主要介绍了脂质载体、聚合物载体、多肽载体及病毒载体,并对未来mRNA药物递送载体研究进行展望,以期为mRNA药物递送新载体的研发提供参考。

槲皮素纳米结构脂质载体的处方与制备工艺优化

目的:对槲皮素纳米结构脂质载体(Q-NLC)的处方与制备工艺进行优化和评价。方法:以粒径大小、多分散指数和Zeta电位等作为质量评价指标,采用单因素试验优化槲皮素纳米结构脂质载体的制备工艺;采用透射电子显微镜观察槲皮素固体脂质纳米粒的微观形态;采用紫外分光光度法测定槲皮素的药物包封率。结果:所制备的Q-NLC为白色乳光的透明乳液,在透射电镜下呈圆形或圆球形状。最优的制备条件为采用溶剂乳化超声法,药物用量为5.0 mg的槲皮素粉末,0.975 g单硬脂酸甘油酯、0.150 g聚乙二醇单硬脂酸酯、0.300 g辛酸癸酸聚乙二醇甘油酯,泊洛沙姆188质量分数为1.0%,纳米结构脂质载体质量浓度为1 mg/mL,制备条件为在400 r/min以及70℃水浴条件下磁力搅拌时间15 min,超声搅拌时间为5 min。槲皮素在0.056~3.12μg/mL范围内线性关系良好,经测定槲皮素固体脂质纳米粒中药物含量为3.12μg/mL,包封率为39.42%±1.12%。结论:经优化制备的Q-NLC分散性好,包封率高,具有良好的稳定性,有望为槲皮素纳米药物的开发和应用提供实验基础。

血清癌胚抗原相关细胞黏附分子1 脂质运载蛋白-2 恶性肿瘤特异生长因子联合检测鉴别诊断乳腺癌的价值

目的 探讨血清癌胚抗原相关细胞黏附分子1(CEACAM1)、脂质运载蛋白2(LCN-2)、恶性肿瘤特异生长因子(TSGF)联合检测鉴别诊断乳腺癌的价值。方法 本研究为回顾性分析,对我院2021年1月至2023年1月收治的40例良性乳腺肿瘤患者临床资料进行收集,作为对照组;并对同期医院收治的40例乳腺癌患者临床资料进行收集,作为观察组。设计基线资料填写表,详细对2组基线资料、血清检查指标进行填写、比较,重点分析血清CEACAM1、LCN-2、TSGF联合检测鉴别诊断乳腺癌的价值。结果 观察组血清CEACAM1、LCN-2、TSGF水平比对照组高,差异有统计学意义(P<0.05),组间年龄、体质指数(BMI)、肿瘤直径、绝经及患侧比较,差异无统计学意义(P>0.05);绘制受试者工作曲线(ROC)结果显示,血清CEACAM1、LCN-2、TSGF检测鉴别诊断乳腺癌的曲线下面积(AUC)均>0.70,且以联合检测(并联)价值最佳。结论 血清CEACAM1、LCN-2、TSGF联合检测鉴别诊断乳腺癌有较高的灵敏度、特异度,鉴别诊断价值满意。

血清AOPP、LCN-2水平在妊娠期糖尿病孕妇不良妊娠结局中的预测分析

目的:研究血清晚期氧化蛋白产物(Gestational diabetes mellitus,AOPP)、人脂质运载蛋白2(Lipocalin 2,LCN-2)水平在妊娠期糖尿病(Gestational diabetes mellitus,GDM)孕妇不良妊娠结局中的预测价值。方法:回顾性收集2021年10月-2022年6月在我院分娩的89例GDM孕妇的临床资料。根据妊娠结局将孕妇分为妊娠结局良好组(n=67)和妊娠结局不良组(n=22)。分析不良妊娠结局情况;对比两组的血清AOPP、LC-2水平;分析血清AOPP、LCN-2水平与GDM孕妇不良妊娠结局的相关性及其对不良妊娠结局的预测价值。结果:妊娠结局不良组血清AOPP、LCN-2较妊娠结局良好组显著提高(P<0.05)。经双变量Pearson分析,血清AOPP、LCN-2水平与GDM孕妇不良妊娠结局呈正相关(r=0.568、0.599,P<0.001)。绘制ROC曲线显示,得到AUC分别为:0.852、0.890、0.915,其中联合检验的预测价值最高。结论:GDM孕妇血清AOPP、LCN-2高表达与不良妊娠结局具有密切关系,且具有一定预测价值。

中性粒细胞明胶酶相关脂质运载蛋白、抗磷脂酶A2受体抗体表达及尿视黄醇结合蛋白水平与慢性肾脏病的关系

目的探讨与分析中性粒细胞明胶酶相关脂质运载蛋白(NGAL)、抗磷脂酶A2受体抗体(PLA2R)表达及尿视黄醇结合蛋白(RBP)水平与慢性肾脏病的关系。方法选择2019年6月—2023年6月在扬州大学附属靖江人民医院诊治的120例慢性肾脏病患者作为研究对象,根据慢性肾脏病定义及经我国人群校正的EPI公式计算得出的估算肾小球滤过率水平将其分为观察组和对照组,将慢性肾脏病Ⅳ期到Ⅴ期的40例患者分为观察组,将Ⅰ期到Ⅲ期的80例患者分为对照组。检测所有患者的血清NGAL、PLA2R表达及RBP水平,检测肾功能常规指标并进行相关性分析。结果观察组的血清NGAL、PLA2R水平均高于对照组,差异有统计学意义(P<0.05)。观察组的RBP水平显著高于对照组,差异有统计学意义(P<0.05),观察组的血清尿素氮、血肌酐水平均显著高于对照组,差异有统计学意义(P<0.05)。在观察组中,Pearson分析显示血清尿素氮、血肌酐水平与NGAL、PLA2R、RBP水平均存在正向相关性,差异有统计学意义(P<0.05)。多因素logistic回归分析显示血清NGAL、PLA2R表达与RBP水平都为导致慢性肾脏病临床分期的重要因素,差异有统计学意义(P<0.05)。结论慢性肾脏病患者多伴随有血清NGAL、PLA2R表达及RBP水平升高,其与患者的常规肾功能指标存在相关性,也可影响慢性肾脏病的进展。

Lipidoid作为新型脂质载体对肝脏非实质细胞的靶向作用实验研究

目的探讨新型脂质载体Lipidoid对肝脏的细胞靶向效率,为抗肝纤维化治疗提供一定实验室依据。方法C57BL/6小鼠分别给予CCl_4灌胃建立肝纤维化模型,同时给予Lipidoid、Lipidoid/siRNA-TIMP-1尾静脉注射,通过Masson染色观察各组小鼠肝脏纤维化情况,并通过实时荧光定量PCR和蛋白免疫印迹检测Lipidoid/siRNA-TIMP-1对TIMP-1表达水平的影响;通过对小鼠尾静脉注射绿色荧光(GFP)标记的Lipidoid-GFP,取其肝组织进行冰冻切片分别进行Desmin及F4/80染色,评价Lipidoid-GFP对肝脏非实质细胞的靶向作用;通过对小鼠分离的原代肝星状细胞SW-HSC、小鼠巨噬细胞系(RAW264.7)转染Lipidoid-GFP,流式细胞仪分别检测上述两种细胞中GFP荧光强度。结果通过对肝组织进行Masson染色发现,在CCl_4诱导的肝纤维化小鼠模型中,给予Lipidoid/siRNA-TIMP-1治疗可显著降低胶原纤维的沉积;实时荧光定量PCR和蛋白免疫印迹结果显示Lipidoid/siRNA-TIMP-1可以显著抑制TIMP-1基因的表达;肝组织冰冻切片Desmin及F4/80染色结果显示,Lipidoid-GFP主要被肝脏肝星状细胞和Kuffer所捕获,肝星状细胞、Kuffer细胞捕获Lipidoid-GFP的阳性率分别为13.4%、43.8%;原代肝星状细胞SW-HSC、小鼠巨噬细胞系(RAW264.7)转染Lipidoid-GFP后经流式细胞仪检测,结果显示转染Lipidoid-GFP的SW-HSC及RAW264.7GFP阳性率分别为66.9%、75.8%,均显著高于对照组。结论小鼠实验结果表明,Lipidoid/siRNA-TIMP-1可有效降低肝脏TIMP-1水平,发挥抗肝纤维化作用。Lipidoid作为一种新型脂质载体可有效感染肝脏非实质细胞(肝星状细胞和Kuffer细胞)。

岩白菜素纳米结构脂质载体制备及其体内药动学研究

目的制备岩白菜素纳米结构脂质载体,并考察其体内药动学。方法熔融法制备纳米结构脂质载体。以固态脂质种类、液态脂质种类、固液脂质比、脂药比、泊洛沙姆188浓度为影响因素,包封率、载药量、粒径为评价指标,单因素试验优化处方,考察体外释药,测定稳定性,进行晶型分析。18只大鼠随机分为3组,分别灌胃给予岩白菜素、物理混合物、岩白菜素纳米结构脂质载体的0.5%CMC-Na混悬液(60 mg/kg),于0.25、0.5、1、2、3、4、5、6、8、10、12 h采血,HPLC法测定岩白菜素血药浓度,计算主要药动学参数。结果最优处方为固态脂质山嵛酸甘油酯,液态脂质油酸,固液脂质比4∶1,脂药比10∶1,泊洛沙姆188浓度2.0%,平均包封率为(84.16±1.57)%,载药量为(7.73±0.27)%,粒径为(215.53±18.04)nm,Zeta电位为-(37.56±2.03)mV。纳米结构脂质载体在模拟胃液中240 min内累积释放度小于50%,在模拟肠液中36 h内为71.04%,并且在后者中12 h内稳定性良好。岩白菜素以无定形状态存在于纳米结构脂质载体中。与原料药、物理混合物比较,纳米结构脂质载体t_(max)、t_(1/2)延长(P<0.01),C_(max)、AUC_(0~t)、AUC_(0~∞)升高(P<0.01),相对生物利用度与原料药相比增加至6.08倍。结论纳米结构脂质载体可改善岩白菜素稳定性、口服生物利用度。