The Effect of N-Acetylation on the Anti-Inflammatory Activity of Chitooligosaccharides and Its Potential for Relieving Endotoxemia.

Endotoxemia is mainly caused by a massive burst of inflammatory cytokines as a result of lipopolysaccharide (LPS) invasion. Chitooligosaccharides (COS) is expected to be a potential drug for relieving endotoxemia due to its anti-inflammatory properties. However, the structural parameters of COS are often ambiguous, and the effect of degree of acetylation (DA) of COS on its anti-inflammatory remains unknown. In this study, four COSs with different DAs (0%, 12%, 50% and 85%) and the same oligomers distribution were successfully obtained. Their structures were confirmed by 1H NMR and MS analysis. Then, the effect of DA on the anti-inflammatory activity and relieving endotoxemia potential of COS was researched. The results revealed that COS with a DA of 12% had better anti-inflammatory activity than COSs with other DAs, mainly in inhibiting LPS-induced inflammatory cytokines burst, down-regulating its mRNA expression and reducing phosphorylation of IκBα. Furthermore, this COS showed an obviously protective effect on endotoxemia mice, such as inhibiting the increase in inflammatory cytokines and transaminases, alleviating the injury of liver and intestinal tissue. This study explored the effect of DA on the anti-inflammatory activity of COS for the first time and lays the foundation for the development of COS as an anti-inflammatory drug against endotoxemia.

Preparation and Antioxidant Activity of Chitosan Dimers with Different Sequences.

As a popular marine saccharide, chitooligosaccharides (COS) has been proven to have good antioxidant activity. Its antioxidant effect is closely related to its degree of polymerization, degree of acetylation and sequence. However, the specific structure-activity relationship remains unclear. In this study, three chitosan dimers with different sequences were obtained by the separation and enzymatic method, and the antioxidant activity of all four chitosan dimers were studied. The effect of COS sequence on its antioxidant activity was revealed for the first time. The amino group at the reducing end plays a vital role in scavenging superoxide radicals and in the reducing power of the chitosan dimer. At the same time, we found that the fully deacetylated chitosan dimer DD showed the strongest DPPH scavenging activity. When the amino groups of the chitosan dimer were acetylated, it showed better activity in scavenging hydroxyl radicals. Research on COS sequences opens up a new path for the study of COS, and is more conducive to the investigation of its mechanism.

Pleiotropic Modulation of Chitooligosaccharides on Inflammatory Signaling in LPS-Induced Macrophages.

Chitooligosaccharide (COS) is a green and non-toxic cationic carbohydrate that has attracted wide attention in recent years due to its anti-inflammatory activity. However, the anti-inflammatory mechanism of COS remains unclear. In this study, RNA-seq was used to investigate the integrated response of COS to LPS-induced damage in macrophages. The results showed that the experimental group with COS had 2570 genes with significant differences compared to the model group, and that these genes were more enriched in inflammatory and immune pathways. The KEGG results showed that COS induces the pleiotropic modulation of classical inflammatory pathways, such as the Toll-like receptor signaling pathway, NF-κB, MAPK, etc. Based on the RNA-seq data and the RT-qPCR, as well as the WB validation, COS can significantly upregulate the expression of membrane receptors, such as Tlr4, Tlr5, and MR, and significantly inhibits the phosphorylation of several important proteins, such as IκB and JNK. Overall, this study offers deep insights into the anti-inflammatory mechanism and lays the foundation for the early application of COS as an anti-inflammatory drug.

Review: Advances in preparation of chitooligosaccharides with heterogeneous sequences and their bioactivity.

Chitooligosaccharides has attracted increasing attention due to their diverse bioactivities and potential application. Previous studies on the bioactivity of chitooligosaccharides were mostly carried out using a mixture. The structure-function relationship of chitooligosaccharides is not clear. Recently, it is confirmed that chitooligosaccharides with different degrees of polymerization play different roles in many bioactivities. However, heterogeneous chitooligosaccharides with a single degree of polymerization is still a mixture of many uncertain sequences and it is difficult to determine which structure is responsible for biological effects. Therefore, an interesting and challenging field of studying chitooligosaccharides with heterogeneous sequences has emerged. Herein, we reviewed the current methods for preparing heterogeneous chitooligosaccharides, including chemical synthesis, separation techniques and enzymatic methods. Advances in the bioactivities of chitooligosaccharides with heterogeneous sequences are also reviewed.