Glycogen Synthase Kinase-3β Mediates Proinflammatory Cytokine Secretion and Adipogenesis in Orbital Fibroblasts from Patients with Graves’ Orbitopathy

Abstract

Purpose We sought to determine the role of glycogen synthase kinase-3β (GSK-3β) in the pathogenesis of Graves’ orbitopathy(GO). Methods Expression of the GSK-3β gene in whole orbital tissue explants was compared between GO and non-GO donors using quantitative real-time PCR (RT-PCR). The expression of proinflammatory molecules in the presence of the GSK-3β inhibitor CHIR 99021 was analyzed using RT-PCR, western blot, and ELISA. Adipogenic differentiation was identified using Oil Red O staining, and the levels of peroxisome proliferator activator gamma (PPARγ) and CCAAT-enhancer-binding proteins (C/EBPs) α and β were determined by western blot. Results The expression of GSK-3β was significantly higher in GO tissues than in control tissues. The addition of CHIR 99021 led to a decrease in the active form of the kinase in which the Y216 residue is phosphorylated. When GO and non-GO fibroblasts were stimulated with IL-1β or TNF-α, IL-6, IL-8, intercellular adhesion molecule-1 (ICAM-1), cyclooxygenase-1 (COX-1), and monocyte chemoattractant protein 1 (MCP-1) showed increased production, which was blunted when CHIR 99021 was added. The activation of Akt, PI3K, nuclear factor (NF)-κB, Erk, Jnk, and p38 kinase by IL-1β and TNF-α was diminished with CHIR 99021 in GO cells. A decrease in lipid droplets and expression of PPARγ and c/EBPα and -β was noted in fibroblasts treated with CHIR 99021 during adipocyte differentiation. The inhibition of Wnt and β-catenin in adipogenesis was reversed by CHIR 99021. Conclusions GSK-3β plays a significant role in GO pathogenesis. The inhibition of the kinase attenuated the proinflammatory cytokines production and fibroblast differentiation into adipocytes. GSK-3β may be a potential target for anti-inflammatory and anti-adipogenic treatment of GO.

Association of fibroblast growth factor 10 with the fibrotic and inflammatory pathogenesis of Graves’ orbitopathy

“…In this study, although basal levels of FGF10 differed between GO and non-GO orbital tissues and primary cells, the inhibitory effect of FGF10 on fibrosis did not differ between GO and non-GO cells. According to previous studies [ 20 , 33 – 35 ], GO and non-GO cells under primary culture conditions often show similar patterns of response to stimulation and inhibition of specific targets that may not reflect actual clinical events. Nonetheless, it is notable that our study found a significant difference in FGF10 expression levels between GO and non-GO tissues and OFs.…”

Glucocorticoids Play a Pivotal Role in Analgesic and Blood Glucose Regulatory Effects in C57 Mice Treated by Water Wxtract of Prunella Vulgaris L. Spica

“…5E). GSK3β was originally found to inactivate glycogen synthase and mediate the development of insulin resistance, then identi ed as a potential target of the therapy for diseases associated with in ammation [9]. In contrast, Akt2/PKB, another GC-target molecule inhibiting glucose release from hepatocytes and increasing insulin sensitivity [29], was also highly up-regulated by 21.41 ± 2.48 folds in PE70 group (Fig.…”

“…Studies using aqueous or organic fraction from P. vulgaris revealed similar effects such as anti-allergy [8], anti-microbial activity [1,2], and anti-in ammation [3]. Aqueous extracts of P. vulgaris (PE) was also reported to inhibit infection [9]. In addition, P. vulgaris is used together with other herbal medicine like Pinellia ternate (Thunb.)…”

Network Pharmacology and ­Experimental Analyses of the Mechanism of Analgesic and Glucose Intolerance Through Glucocorticoid Signaling in C57 Mice Treated with Water Extract of Prunella vulgaris L. Spica

“…GSK3β was originally found to inactivate glycogen synthase and mediate the development of insulin resistance, then identified as a potential target of the therapy for diseases associated with inflammation. 9 In contrast, Akt2/PKB, another GC-target molecule inhibiting glucose release from hepatocytes and increasing insulin sensitivity, 37 was also highly up-regulated by 21.41 ± 2.48 folds in the PE70 group (Figure 6F).…”

“…3 Aqueous extracts of PE were also reported to inhibit infection. 9 In addition, P. vulgaris is used, together with other herbal medicines, like Pinellia ternata Thunb., for some classic decoctions in Chinese medicine for dispersing hard lumps and abscesses in tissues, 10 or as an over-the-counter medication for hypertension. 11 For example, Xia Sang Ju formula, a Chinese drug and herbal tea made up of Prunellae spica, Mori folium, and Flos Chrysanthemi Indici, has commonly been used in the clinic for fever, headache, and sore throat since 1814.…”