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口服纳米载体可以有效地提高难溶性药物的生物利用度,其与肠上皮细胞的相互作用研究也成为一大热点。在该领域的研究中,物理包载或化学偶联荧光基团的纳米载体常常被选择,但是物理包载荧光染料可能会发生泄漏,影响结果分析,而化学偶联荧光探针可能会改变纳米粒结构的表面特征。因此,需要寻找一种简便、准确的检测方法以探究口服纳米粒内吞及跨膜转运的过程、胞内分布和机制。本文选择金纳米粒为纳米模型载体,基于金纳米粒具有光散射的性质,以633 nm波长的激光为激发光源,通过激光扫描共焦显微镜(CLSM)检测金纳米粒溶液,肠道上皮细胞内及大鼠肠道中的金纳米粒。结果通过激光共聚焦显微镜反射光的模式,无论在细胞水平还是大鼠肠道中,都能够准确地检测到金纳米粒的信号,进而分析金纳米粒入胞及跨越肠道上皮细胞的过程和胞内分布,且不受背景的干扰,并能借助层扫模式,构建金纳米粒在细胞内分布的3D模型。因此,在不进行荧光探针偶联的条件下,通过反射光分析金纳米粒跨膜转运是准确、简便的方法。
Oral nano-carriers can effectively improve the bioavailability of poorly soluble drugs, and its interaction with intestinal epithelial cells has also become a hot topic. In this area of study, nanocarriers that are physically or chemically conjugated to fluorophores are often selected, but physical inclusion of fluorescent dyes can leak and affect the result analysis, whereas chemically coupled fluorescent probes can change the nanometer Surface features of grain structure. Therefore, there is a need to find a simple and accurate detection method to investigate the process, intracellular distribution and mechanism of oral nanoparticle endocytosis and transmembrane transport. In this paper, gold nanoparticle is selected as the carrier of nanometer model. Based on the light scattering properties of gold nanoparticle, the gold nanoparticle solution is detected by laser scanning confocal microscopy (CLSM) with 633 nm wavelength laser as excitation light source. And gold nanoparticles in rat intestine. Results The pattern of light reflected by confocal laser scanning microscope could detect the signal of gold nanoparticles accurately at the cellular level and in the intestine of rat and then analyze the process of entering and crossing the epithelial cells of gold particles Within the distribution, and not subject to background interference, and can use the layer sweep mode to build a 3D model of gold nanoparticles distributed in the cell. Therefore, it is an accurate and simple method to analyze transmembrane transport of gold nanoparticles by reflected light without fluorescent probe coupling.