METHODS: High pressure homogenization methods were used to fabricate oleic acid embedded lipid nanoparticles (OALNs). The in vitro and in vivo models were used to validate the physiological effect of this OALNs via various cellular and molecular approaches including cell viability essay, fluorescent staining, electron microscope, RNAseq, qPCR, Western blots, and IHC staining.
RESULTS: We successfully fabricated OALNs with enhanced stability and solubility. More importantly, lipid accumulation was successfully induced in hepatocytes via the application of OALNs in a dose-dependent manner. Overload of OALNs resulted in ROS accumulation and apoptosis of hepatocytes dose-dependently. With the help of transcriptome sequencing and traditional experimental approaches, we demonstrated that the lipotoxic effect induced by OALNs was exerted via the DDIT3/BCL2/BAX/Caspases signaling. Moreover, we also verified that OALNs induced steatosis and subsequent apoptosis in the liver of mice via the activation of DDIT3 in vivo.
CONCLUSIONS: In all, our results established a potential pathogenic model of NAFLD for further studies and indicated the possible involvement of DDIT3 signaling in abnormal steatosis process of the liver.
方法:使用高压均质方法制备油酸包埋的脂质纳米颗粒(OALN)。体外和体内模型用于通过各种细胞和分子方法验证这种OALN的生理效应,包括细胞活力论文。荧光染色,电子显微镜,RNAseq,qPCR,西方印迹,和IHC染色。
结果:我们成功地制造了具有增强的稳定性和溶解性的OALN。更重要的是,通过以剂量依赖的方式应用OALN成功诱导肝细胞中的脂质积累。OALN的过载导致ROS积累和肝细胞的凋亡呈剂量依赖性。在转录组测序和传统实验方法的帮助下,我们证明了OALN诱导的脂毒性作用是通过DDIT3/BCL2/BAX/Caspases信号传导产生的。此外,我们还证实OALN通过体内DDIT3的激活诱导小鼠肝脏脂肪变性和随后的细胞凋亡。
结论:总而言之,我们的结果为进一步研究建立了NAFLD的潜在致病模型,并表明DDIT3信号传导可能参与肝脏异常脂肪变性过程.