Pauloin Thierry, Dutot Mélody, Warnet Jean-Michel, Rat Patrice
Laboratoire de Toxicologie, Faculté des Sciences Pharmaceutiques et Biologiques, Université Paris Descartes, 75006 Paris, France.
Eur J Pharm Sci. 2008 Aug 7;34(4-5):263-73. doi: 10.1016/j.ejps.2008.04.006. Epub 2008 May 1.
Benzalkonium chloride (BAK) is one of the most often used preservative in pharmaceutical products and it is known to induce toxic effects. Hyaluronan (HA), a linear biopolymer, is involved in several biological processes. The aim of this work is to in vitro investigate if HA is able to decrease BAK toxicity.
Two human epithelial cell lines were treated with different incubation time protocol with BAK and three different molecular weights HA (HA 20k Da, HA 100 kDa and HA 1000 kDa, 0.2%, w/v). Flow cytometry, fluorescence microscopy, microplate cytofluorometry and confocal microscopy were performed to evaluate expression of CD44 receptor, cell viability, oxidative stress, mitochondrial mass, chromatin condensation, plasma-membrane permeability, DNA fragmentation and cytoskeleton morphology.
The three HAs studied induce neither oxidative stress nor apoptosis. HA 1000 kDa significantly decreases oxidative stress, apoptosis and necrosis induced by BAK. Experiments with HA 20 kDa or HA 100 kDa did not show the same effects. For instance, the more molecular weight decreases, the more protection decreases. Moreover, we suggest that HA interacts with cell plasma-membrane and inhibits cell death receptors.
High molecular weight HA (1000 kDa, 0.2%) is an effective protective agent against BAK.
苯扎氯铵（BAK）是药品中最常用的防腐剂之一，已知其具有毒性作用。透明质酸（HA）是一种线性生物聚合物，参与多种生物过程。本研究旨在体外探究HA是否能够降低BAK的毒性。
用BAK和三种不同分子量的HA（HA 20kDa、HA 100kDa和HA 1000kDa，0.2%，w/v）按照不同的孵育时间方案处理两种人类上皮细胞系。采用流式细胞术、荧光显微镜、微孔板细胞荧光测定法和共聚焦显微镜来评估CD44受体的表达、细胞活力、氧化应激、线粒体质量、染色质凝聚、质膜通透性、DNA片段化和细胞骨架形态。
所研究的三种HA均未诱导氧化应激或凋亡。HA 1000kDa显著降低了BAK诱导的氧化应激、凋亡和坏死。HA 20kDa或HA 100kDa的实验未显示出相同的效果。例如，分子量越低，保护作用越弱。此外，我们认为HA与细胞质膜相互作用并抑制细胞死亡受体。
高分子量HA（1000kDa，0.2%）是一种有效的抗BAK保护剂。
