背景:大黄素是一种在传统中草药中发现的化合物。它具有抗炎和许多其他药理作用。我们先前的研究表明,大黄素可显着减轻重症急性胰腺炎(SAP)的炎症作用。然而,其溶解性差,高毒性和有限的胰腺保留时间阻碍了其临床应用。
目的:我们旨在制备具有改善的生物利用度的大黄素纳米胶囊,以通过靶向巨噬细胞实现大黄素的控释。Further,探讨了负载大黄素的甘露糖偶联壳聚糖包被的脂质纳米胶囊(M-CS-E-LNC)治疗SAP的机制。
方法:通过相转化法制备了M-CS-E-LNC,并进行了少量修饰。ELISA法检测M-CS-E-LNC的炎症介质表达和抗炎作用,巨噬细胞和LPS诱导的SAP小鼠中的IHC和IF。应用IVIS光谱成像和HPLC探索M-CS-E-LNC在胰腺中的控释。进行LC-MS/MS用于巨噬细胞的脂质组学分析。此外,基于载体的短发夹RNA(shRNA)方法用于沉默巨噬细胞中CTP1基因的表达.
结果:用M-CS-E-LNC治疗后,巨噬细胞中炎症介质的水平明显降低。通过对血清淀粉酶水平的分析,在SAP小鼠中检测到相同的抗炎作用,TNF-α和IL-6。重要的是,M-CS-E-LNC允许大黄素选择性地积聚在胰腺和胃肠道组织,从而表现出针对性的释放。机械上,M-CS-E-LNC治疗组显示肉碱棕榈酰转移酶1(CPT1)蛋白的表达上调,促进细胞内长链脂肪酸的转运,从而促进巨噬细胞的M2表型极化。
结论:M-CS-E-LNC具有显著改善的生物利用度和水溶性,这对巨噬细胞极化有更大的治疗作用。我们的发现还表明,第一次,CPT1可能是SAP治疗的新治疗靶点。
BACKGROUND: Emodin is a chemical compound found in traditional Chinese herbs. It possesses anti-inflammatory and many other pharmacological effects. Our previous study showed that emodin significantly alleviates the inflammation effect of severe acute pancreatitis (SAP). However, its poor solubility, high toxicity and limited pancreas retention time hinder its clinical application.
OBJECTIVE: We aimed to prepare emodin
nanocapsules with improved bioavailability to achieve the controlled release of emodin by targeting macrophages. Further, the mechanism of mannose-conjugated chitosan-coated lipid
nanocapsules loaded with emodin (M-CS-E-LNC) in the treatment of SAP was explored.
METHODS: M-CS-E-LNC were prepared by the phase inversion method with slight modification. The expression of inflammation mediators and the anti-inflammation efficacy of M-CS-E-LNC were examined by ELISA, IHC and IF in macrophage cells and LPS-induced SAP mice. IVIS spectrum imaging and HPLC were applied to explore the controlled release of M-CS-E-LNC in the pancreas. LC-MS/MS was performed for lipidomics analysis of macrophages. Moreover, a vector-based short hairpin RNA (shRNA) method was used to silence CTP1 gene expression in macrophage cells.
RESULTS: The levels of inflammatory mediators in macrophages were markedly decreased after treatment with M-CS-E-LNC. The same anti-inflammation effects were detected in SAP mouse through the analysis of serum levels of amylase, TNF-α and IL-6. Importantly, M-CS-E-LNC allowed the emodin to selectively accumulate at pancreas and gastrointestinal tissues, thus exhibiting a targeted release. Mechanistically, the M-CS-E-LNC treatment group showed up-regulated expression of the carnitine palmitoyltransferase 1 (CPT1) protein which promoted intracellular long-chain fatty acid transport, thereby promoting the M2 phenotype polarization of macrophages.
CONCLUSIONS: M-CS-E-LNC exhibited significantly improved bioavailability and water solubility, which translated to greater therapeutic effects on macrophage polarization. Our findings also demonstrate, for the first time, that CPT1 may be a new therapeutic target for SAP treatment.