Protection effect of baicalin-regulated IKKα on inflammatory reaction of human oral keratinocytes

doi:10.19439/j.sjos.2021.06.006

Abstract

Abstract: PURPOSE: To observe the regulation of baicalin on IKKα mediated MASPIN in Human oral keratinocytes (HOKs) inflammatory reaction, this study was to explore the molecular regulation mechanism of baicalin on oral mucosal inflammation. METHODS: HOKs were stimulated by lipopolysaccharide (LPS) to mimic the inflammatory response of oral mucosal inflammation in vitro. CCK-8 assay was used to detect the toxicity of baicalin to HOKs; then different concentrations of baicalin were pre-treated to LPS-stimulated HOKs, enzyme-linked immunosorbent assays (ELISA) was used to detect the secretion of IL-6 and TNF-α in LPS-stimulated HOKs; reverse transcription polymerase chain reaction(RT-PCR) and Western blot assay were used to detect the regulatory effects of baicalin on gene and protein expression levels of IKKα mediated MASPIN in LPS-stimulated HOKs. SPSS 21.0 software package was used for statistical analysis of the data. RESULTS: HOKs stimulated by 10 μg/mL LPS successfully simulated the inflammatory environment of oral mucosal inflammation. The concentration of baicalin between 1 μg/mL and 20 μg/mL had no toxic effect on HOKs. With the increasing concentration of baicalin, the expression of MASPIN increased gradually, while the expression of IKKα and inflammatory factors decreased gradually(P<0.05). CONCLUSIONS: Baicalin can decrease the expression of inflammatory factors in LPS-stimulated HOKs, down-regulate IKKα and up-regulate MASPIN.

Key words: Baicalin, Mammary serine protease inhibitor, IKKα, Oral mucosal inflammation

CLC Number:

R781.5

LIU Zhuan-zhuan, XU Na, GAO Rui-fang, ZHANG Fang. Protection effect of baicalin-regulated IKKα on inflammatory reaction of human oral keratinocytes[J]. Shanghai Journal of Stomatology, 2021, 30(6): 589-594.

References

[1] Li C, Tang X, Zheng X, et al.Global prevalence and incidence estimates of oral lichen planus: a systematic review and meta-analysis[J]. JAMA Dermatol, 2020, 156(2): 172-181.
[2] Gueiros L, Arao T, Souza T, et al.IL17A polymorphism and elevated IL17A serum levels are associated with oral lichen planus[J]. Oral Dis, 2018, 24(3): 377-383.
[3] Kara YA.The measurement of serum tumor necrosis factor-alpha levels in patients with lichen planus[J]. Indian J Dermatol, 2018, 63(4): 297-300.
[4] Mozaffari HR, Sharifi R, Sadeghi M.Interleukin-6 levels in the serum and saliva of patients with oral lichen planus compared with healthy controls: a meta-analysis study[J]. Cent Eur J Immunol, 2018, 43(1): 103-108.
[5] Gao Y, Liu H, Wang H, et al.Baicalin inhibits breast cancer development via inhibiting IkB kinase activation in vitro and in vivo[J]. Int J Oncol, 2018, 53(6): 2727-2736.
[6] Tao Y, Zhan S, Wang Y, et al.Baicalin, the major component of traditional Chinese medicine Scutellaria baicalensis induces colon cancer cell apoptosis through inhibition of oncomiRNAs[J]. Sci Rep, 2018, 8(1): 14477.
[7] Wang Q, Xu H, Zhao X.Baicalin inhibits human cervical cancer cells by suppressing protein kinase c/signal transducer and activator of transcription (pkc/stat3) signaling pathway[J]. Med Sci Monit, 2018, 24:1955-1961.
[8] Wang H, Li H, Chen F, et al.Baicalin extracted from huangqin (radix scutellariae baicalensis) induces apoptosis in gastric cancer cells by regulating b cell lymphoma (bcl-2)/bcl-2-associated x protein and activating caspase-3 and caspase-9[J]. J Tradit Chin Med, 2017, 37(2): 229-235.
[9] Yu Y, Pei M, Li L.Baicalin induces apoptosis in hepatic cancer cells in vitro and suppresses tumor growth in vivo[J]. Int J Clin Exp Med, 2015, 8(6): 8958-8967.
[10] Wang C, Song Y, Wang X, et al.Baicalin ameliorates collagen-Induced arthritis through the suppression of janus kinase 1 (JAK1)/signal transducer and activator of transcription 3 (STAT3) signaling in mice[J]. Med Sci Monit, 2018, 24: 9213-9222.
[11] Yu F, Xu N, Zhou Y, et al.Anti‐inflammatory effect of paeoniflorin combined with baicalin in oral inflammatory diseases[J]. Oral Dis, 2019, 25(8): 1945-1953.
[12] Zou Z, Anisowicz A, Hendrix M, et al.Maspin, a serpin with tumor-suppressing activity in human mammary epithelial cells[J]. Science, 1994, 263(5146): 526-529.
[13] Xu N, Li B, Liu Z, et al.Role of mammary serine protease inhibitor on the inflammatory response in oral lichen planus[J]. Oral Dis, 2019, 25(4): 1091-1099.
[14] Cao T, Zhang H, Zhou L, et al.In vitro cell culture system optimization of keratinocytes from oral lichen planus(OLP) patients[J]. Oral Dis, 2016, 23(2): 225-232.
[15] Wang J, Luo H, Yang L, et al.Baicalein induces apoptosis and reduces infammation in LPS-stimulated keratinocytes by blocking the activation of NF-κB: implications for alleviating oral lichen planus[J]. Cell Mol Biol (Noisy-le-grand), 2016, 62(7): 55-60.
[16] Sun JY, Li DL, Dong Y, et al.Baicalin inhibits toll-like receptor 2/4 expression and downstream signaling in rat experimental periodontitis[J]. Int Immunopharmacol, 2016, 36(1): 86-93.
[17] Wang G, Mohammadtursun N, Lv Y, et al.Baicalin exerts anti-airway inflammation and anti-remodelling effects in severe stage rat model of chronic obstructive pulmonary disease[J]. Evid Based Complement Alternat Med, 2018, 2018: 7591348.
[18] Yang X, Zhang Q, Gao Z, et al.Baicalin alleviates IL-1β-induced inflammatory injury via down-regulating mir-126 in chondrocytes[J]. Biomed Pharmacother, 2018, 99:184-190.
[19] Sun SC.The non-canonical NF-κB pathway in immunity and inflammation[J]. Nat Rev Immunol, 2017,17(9): 545-558.
[20] Ito M, Hamano T, Komatsu T, et al.A novel IKKα inhibitor, noraristeromycin, blocks the chronic inflammation associated with collagen-induced arthritis in mice[J]. Mod Rheumatol, 2014, 24(5):775-780.
[21] Zhao Y, Wang HL, Li TT, et al.Baicalin ameliorates dexamethasone-induced osteoporosis by regulation of the RANK/RANKL/OPG signaling pathway[J]. Drug Des Devel Ther, 2020,15(14):195-206.
[22] Wang N, Chang LL.The potential function of IKKα in gastric precancerous lesion via mediating Maspin[J]. Tissue Cell, 2020, 65:101349.
[23] Luo JL, Tan W, Ricono JM, et al.Nuclear cytokine-activated IKKa controls prostate cancer metastasis by repressing Maspin[J]. Nature, 2007, 446(7136): 690-694.
[24] Leopizzi M, Cocchiola R, Milanetti E, et al.IKKa inhibition by a glucosamine derivative enhances maspin expression in osteosarcoma cell line[J]. Chem Biol Interact, 2017, 262(1):19-28.
[25] Lsci Bostanci E, Guler I, Dikmen AU, et al.Prognostic role of maspin expression in patients with cervical dysplasia and cervical cancer[J]. J Obstet Gynaecol Res, 2020,46(5):759-764.
[26] Toll A, Margalef P, Masferrer E, et al.Active nuclear IKK correlates with metastatic risk in cutaneous squamous cell carcinoma[J]. Arch Dermatol Res, 2015, 307(8): 721-729.
[27] Choi KY, Choi HJ, Chung EJ, et al.Loss of heterozygosity in mammary serine protease inhibitor (maspin) and p53 at chromosome 17 and 18 in oral cavity squamous cell carcinoma[J]. Head Neck, 2015, 37(9): 1239-1245.