Baicalin Regulates MASPIN And IKK? Expression Of HOKs Through NF-?B Non-classical Pathway

Objectives:Baicalin from the root of Scutellaria baicalensis Georgi has anti-inflammatory properties.Previous studies have found that the mammary serine protease inhibitor(MASPIN)mediated by I?B kinase(IKK kinase)is involved in the inflammatory process of oral lichen planus(OLP).The purpose of this study is to explore the inhibitory effect of baicalin on the inflammatory response of Lipopolysaccharide(LPS)stimulated HOKs and the molecular mechanism of MASPIN and IKK? regulation via NF-?B non-classical pathway,and to provide a new research idea for the drug treatment of oral mucosal inflammatory diseases such as oral lichus planus(OLP).Materials and Methods:Stimulating human oral mucosal keratinocytes(HOKs)with 10?g/ml LPS to simulate the inflammatory environment of oral lichen planus in vitro.Cell proliferation toxicity assay CCK-8 was used to detect the toxic effects of baicalin in different concentrations on HOKs;HOKs were pre-treated with baicalin in different concentrations,Enzyme?linked immunosorbent assays(ELISA)were used to detect the secretion of IL-6 and TNF-? in LPS-stimulated HOKs;Reverse transcription polymerase chain reaction(RT-PCR)assays and Western blot were used to detect the regulatory effects of baicalin on the gene and protein expression levels of MASPIN and IKK? in LPS-stimulated HOKs.In order to further study the molecular mechanism of baicalin regulating MASPIN expression in HOKs,NF-?B inhibitor(BAY 11-7082)was pre-added to the LPS-stimulated HOKs,The toxic effects of baicalin on HOKs and the expression levels of MASPIN and IKK? genes and proteins in HOKs stimulated by LPS ofpre-added inhibitor(BAY 11-7082)as well as the regulatory effects on the secretion of IL-6 and TNF-? were detected.Results:HOKs stimulated by 10?g/ml LPS can successfully simulate the inflammatory environment of OLP in vitro.The concentration of baicalin between 1?g/ml and 20?g/ml had no toxic effect on HOKs.With the increasing concentration of baicalin,the expression of MASPIN increased gradually,while the expression of IKK? and inflammatory factors decreased gradually(P < 0.05).NF-?B inhibitor(BAY 11-7082)(0.1-5?M)has no toxic effect on HOKs,When the concentration of inhibitor is 2.5?M,it can significantly inhibit the production of IKK?,IL-6,and TNF-? in the HOKs stimulated by LPS and promoted the expression of MASPIN.The regulatory effect of baicalin on HOKs related factors is similar to that of inhibitors.After the addition of NF-?B inhibitors(BAY 11-7082)in LPS-stimulated HOKs,The concentration dependence of baicalin on the inhibition of inflammatory factors disappeared.there was no statistically significant difference in the expression levels of genes and protein of MASPIN?IKK??IL-6 and TNF-? in LPS-stimulated HOKs compared with the group without inhibitors.NF-?B inhibitors block the anti-inflammatory effects of baicalin.Conclusion:Baicalin may promote MASPIN expression through the NF-?B non-classical pathway and inhibit the inflammatory response of LPS-stimulated HOKs,providing a new research idea for the drug treatment of oral mucosal inflammatory diseases such as OLP.