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.