Protective Effect Of Three Active Components Of Traditional Chinese Medicine On Inflammatory Injury

Inflammation is the key response of the body to resist foreign influences and maintain the homeostasis of the body,and it is also the early symptom of various diseases.Chinese herbs and their monomers have the advantages of wide sources,low toxicity,low resistance,and immunity enhancement,which have become natural resources for the research and development of new anti-inflammatory drugs.Baicalein appears in the compatibility of various classical compounds due to its wide range of pharmacological activities,and baicalein is the main active component of the traditional Chinese medicine Scutellaria baicalein.Dihydromyricetin is the main active ingredient of vine tea,which has a long drinking history,and ursolic acid is the main active ingredient of many traditional Chinese medicines.In this study,we studied the protective effects of baicalin,dihydromyricetin and ursolic acid on lipopolysaccharide(LPS)-induced inflammation in mice and the underlying mechanisms,to provide a basis for the development of anti-inflammatory drugs and the study of the basic mechanism of traditional Chinese medicine.Lipopolysaccharide(LPS)was used to stimulate mouse macrophage RAW264.7cells to establish an inflammatory injury model in vitro.The effects of baicalin,dihydromyricetin and ursolic acid on the phagocytosis and proliferation of macrophages were measured,and the mRNA expression of nitric oxide and pro-inflammatory cytokines iNOS,IL-1,IL-6 and TNF-α were measured.The anti-inflammatory effects of three TCM active ingredients were verified and compared.Animal models of inflammation were established by intragastric administration of baicalin,dihydromyricetin and ursolic acid at different concentrations for 7 consecutive days followed by intraperitoneal injection of lipopolysaccharide.The changes of spleen organ coefficient were measured.The differentiation and proliferation of CD3+,CD4+and CD8+ T cells were detected by flow cytometry.Real-time PCR was used to detect the gene expression of inflammatory factors IL-1 and TNF-α to verify the anti-inflammatory effect of the three active components of traditional Chinese medicine in vivo.The proliferation and differentiation of Th17 and Treg cells in the spleen were detected by flow cytometry.The mrna expression levels of IL17,IL-22,IL-10 and FoxP3 in macrophages and spleen tissues were detected by real-time PCR.The gene expressions of JAK/STAT and TGF/SMAD signaling pathway related factors in macrophages and spleen tissues were detected by real-time PCR to verify the mechanism.Findings were as follows:1.The results of in vitro experiments showed that baicalin,dihydromyricetin and ursolic acid could increase the phagocytic capacity of macrophages,promote the proliferation of macrophages,inhibit the expression of inflammatory factors,and have a good protective effect on LPS-induced inflammatory injury.At the same dose concentration of 6.25 μg/m L,dihydromyricetin had the best anti-inflammatory activity.2.Baicalin,dihydromyricetin and ursolic acid could reduce the swelling of mouse spleen,and then reduce the change of spleen organ coefficient.In addition,the three active ingredients of traditional Chinese medicine could promote the proliferation and differentiation of T cells in spleen tissue,and restore the balance of CD4/CD8 by promoting the differentiation of CD4+ cells and inhibiting the proliferation of CD8+cells,thus playing an anti-inflammatory role.3.Baicalin,dihydromyricetin and ursolic acid can promote the proliferation and differentiation of Treg cells in T cells,and inhibit the differentiation of Th17 cells.The mRNA expression of IL-17,which is associated with Th17/Treg differentiation,was significantly inhibited,while the expression of FoxP3 was enhanced in macrophages and spleen tissues,which were consistent with the results of flow cytometry.4.LPS could activate the expression of JAK/STAT signaling pathway and inhibit the activation of TGF/SMAD signaling pathway.Baicalin,dihydromyricetin and ursolic acid all had the ability to reverse this result.Baicalin,dihydromyricetin and ursolic acid may inhibit the activation of JAK1/3-STAT1/6,JAK1/3-STAT1/6 and JAK1/2-STAT6 signaling pathways in macrophages in vitro.Baicalin may inhibit the activation of JAK-STAT1/3 pathway,dihydromyricetin may inhibit the activation of JAK2/3-STAT3/6 pathway,and ursolic acid may inhibit the activation of JAK2/3-STAT1/6 pathway in mice.Taken together,baicalin,dihydromyricetin and ursolic acid have protective effects on LPS-induced inflammation,which may be related to inhibiting JAK/STAT signaling pathway,activating TGFβ/SMAD signaling pathway,inhibiting the expression of inflammatory factors,and promoting the Th17/Treg balance axis.