Attenuation of obesity related inflammation in RAW 264.7 macrophages and 3T3-L1 adipocytes by varanadi kashayam and identification of potential bioactive molecules by UHPLC-Q-Orbitrap HRMS

Obesity is associated with chronic low-grade inflammation, characterised by the up-regulation of pro-inflammatory cytokines in obese adipose tissue. In this study, 3T3-L1 CM enhanced TNF-α and IL-1β in RAW 264.7 cells compared with LPS treated cells. However, treatment with Varanadi kashayam suppressed the inflammatory changes associated with RAW 264.7 cells. Subsequently, RAW CM used to stimulate adipocytes, resulting in decreased intracellular lipid content and reduced adipogenic markers after Varanadi kashayam treatment. The chemical profiling of Varanadi kashayam using UHPLC-Q-Orbitrap-HRMS identified 194 compounds by comparing their retention time, the experimentally measured exact mass of precursor, and fragmented ions, and fragmentation pattern with spectral library and reported literature. Collectively, Varanadi kashayam act as a potent anti-inflammatory and anti-adipogenic agent that could disrupt the crosstalk between adipocytes and macrophages. Hence it could be a better candidate for reducing inflammation associated with obese adipose tissue.

肥胖与慢性低度炎症密切相关，该炎症以肥胖脂肪组织中促炎细胞因子的上调为典型特征。本研究中，与脂多糖（Lipopolysaccharide, LPS）处理组相比，3T3-L1条件培养基（Conditioned Medium, CM）可上调RAW 264.7细胞内肿瘤坏死因子-α（TNF-α）与白细胞介素-1β（IL-1β）的表达水平。然而，经Varanadi kashayam处理后，RAW 264.7细胞的炎症相关病理改变得到显著抑制。后续实验中，研究人员采用RAW 264.7条件培养基刺激脂肪细胞；结果显示，经Varanadi kashayam处理后，细胞内脂质含量降低，成脂标志物的表达水平也有所下调。本研究采用超高效液相色谱-四极杆-轨道阱高分辨质谱（Ultra High Performance Liquid Chromatography-Quadrupole-Orbitrap High Resolution Mass Spectrometry, UHPLC-Q-Orbitrap-HRMS）对Varanadi kashayam进行化学成分分析，通过比对各化合物的保留时间、前体离子实测精确质量、碎片离子及碎裂模式与光谱数据库及已发表文献数据，共鉴定出194种成分。综上，Varanadi kashayam是一种强效的抗炎及抗成脂制剂，可干扰脂肪细胞与巨噬细胞间的信号串扰，因此有望成为缓解肥胖相关脂肪组织炎症的优良候选药物。