Extraction technology, component analysis, and the immunomodulatory effects in immunosuppressed broilers of total flavonoids extract from the thorns of Gleditsia sinensis Lam

Spina Gleditsiae is the dry thorns from Gleditsia sinensis Lam. (G. sinensis), exhibits the effect of reducing swelling and toxins, and draining pus. Flavonoids are the primary active constituents of Spina Gleditsiae, but their extraction technology has not been studied systematically. Relevant studies revealed that Spina Gleditsiae shown certain immunomodulatory effects, but its mechanism was still unclear. This research was carried out about the extraction process, chemical composition, and the effects on the immune function of total flavonoids extract (TFE). The extraction conditions of TFE were investigated by response surface methodology, and the major components were preliminarily analyzed and inferred by ultra performance liquid chromatography- quadrupole-time-of-flight-mass spectrum method. The immunomodulatory effect of TFE was evaluated by cyclophosphamide induced immunosuppressed broilers. The yield of TFE was 1.80 % under the following optimized conditions: ethanol concentration 50 %, extraction time 20 min, and the ratio of solvent to material 50:1 ml/g. In addition, the main chemical components in the TFE have been preliminarily identified based on related literature and mass spectrometry information. Meanwhile, the TFE could mitigate the immunosuppressive state caused by cyclophosphamide in broilers by improving the histomorphology of immune organs, increasing the index of immune organ, elevating the serum levels of IL-2, IFN-β, and IFN-γ, and raising the titer of Newcastle disease antibodies in the serum. Furthermore, the immunoregulatory effects of these flavonoids were found to be closely related to the enhancement of the relative expression level of key genes and proteins in the TLR4-MyD88/TICAM-NF-κB signaling pathway. The findings of the study suggest that TFE derived from the thorns of G. sinensis holds promise as an enhanced traditional Chinese medicine with the capability to potentially bolster immune function.