School and Hospital of Stomatology, China Medical University, Shenyang, China.
J Periodontal Res. 2024 Aug;59(4):812-820. doi: 10.1111/jre.13254. Epub 2024 May 3.
To investigate whether visceral adipose tissue-derived serine protease inhibitor (vaspin) can alleviate the inhibitory effect of high-glucose (HG) culture on the proliferation and osteogenic differentiation of human periodontal ligament stem cells (PDLSCs) and to preliminarily explore the underlying mechanisms.
High glucose produces damage to the regeneration of periodontal tissue of PDLSCs. The expression level of vaspin in periodontal tissue is high in periodontitis patients and effectively reduced after initial therapy of periodontal diseases. However, the effect of vaspin on PDLSCs remains unknown.
PDLSCs were cultured in media augmented with 5.5 or 25.0 mM concentrations of glucose to elucidate the impact and mechanism of vaspin on PDLSCs under high glucose in vitro. Proliferation was measured by Cell Counting Kit-8 (CCK8) assay. Osteogenesis of PDLSCs was assessed by alkaline phosphatase (ALP) staining, ALP activity, and Alizarin Red staining. Quantitative real-time polymerase chain reaction (qRT-PCR) and western blot (WB) were used to investigate the osteo-specific markers. Then, the molecular impact of vaspin in the presence/absence of HG on PDLSCs physiology was determined with TGF-β1/Smad signaling pathway as the main focus.
It was revealed that the proliferation and osteogenic differentiation (OD) of PDLSCs under HG was reduced, and by adding vaspin the anti-osteogenic impact of HG was relieved. Moreover, vaspin enhanced TGF-β1/Smad signaling pathway activity. Pretreatment with TGF-β1 inhibitor blocked vaspin-triggered TGF-β1/Smad signal activation and minimized the vaspin-induced protective effect against HG-inhibited growth and OD.
In summary, vaspin observably reduces HG-mediated inhibition of PDLSCs OD by modulating the TGF-β1/Smad signaling pathway. Vaspin may be a potential therapeutic for periodontal tissue regeneration in diabetic patients.
探讨内脏脂肪组织来源的丝氨酸蛋白酶抑制剂(vaspin)是否可以减轻高糖(HG)培养对人牙周膜干细胞(PDLSCs)增殖和成骨分化的抑制作用,并初步探讨其潜在机制。
高糖对 PDLSCs 牙周组织再生造成损害。在牙周炎患者的牙周组织中,vaspin 的表达水平较高,在牙周病的初始治疗后有效降低。然而,vaspin 对 PDLSCs 的影响尚不清楚。
用含有 5.5 或 25.0 mmol/L 葡萄糖的培养基培养 PDLSCs,以阐明高糖环境下 vaspin 对 PDLSCs 的影响及其机制。通过 Cell Counting Kit-8(CCK8)测定法测量增殖。通过碱性磷酸酶(ALP)染色、ALP 活性和茜素红染色评估 PDLSCs 的成骨分化。采用定量实时聚合酶链反应(qRT-PCR)和蛋白质印迹(WB)检测成骨特异性标志物。然后,以 TGF-β1/Smad 信号通路为主要关注点,确定 vaspin 在 HG 存在/不存在的情况下对 PDLSCs 生理的分子影响。
结果表明,HG 下 PDLSCs 的增殖和成骨分化(OD)降低,添加 vaspin 可减轻 HG 对成骨的抑制作用。此外,vaspin 增强了 TGF-β1/Smad 信号通路活性。TGF-β1 抑制剂预处理阻断了 vaspin 触发的 TGF-β1/Smad 信号激活,并使 vaspin 诱导的对 HG 抑制生长和 OD 的保护作用最小化。
总之,vaspin 通过调节 TGF-β1/Smad 信号通路显著降低 HG 介导的 PDLSCs OD 抑制。vaspin 可能是糖尿病患者牙周组织再生的潜在治疗方法。