Department of Chemistry and Biochemistry, Laboratory of Bioassays and Cell Dynamics, Institute of Biosciences, Sao Paulo State University - UNESP, Botucatu, São Paulo, Brazil.
Department of Restorative Dentistry, School of Dentistry at Araraquara, Sao Paulo State University - UNESP, Araraquara, São Paulo, Brazil.
J Mater Sci Mater Med. 2021 Mar 22;32(4):33. doi: 10.1007/s10856-021-06504-y.
Phytocystatins are endogenous cysteine-protease inhibitors present in plants. They are involved in initial germination rates and in plant defense mechanisms against phytopathogens. Recently, a new phytocystatin derived from sweet orange, CsinCPI-2, has been shown to inhibit the enzymatic activity of human cathepsins, presenting anti-inflammatory potential and pro-osteogenic effect in human dental pulp cells. The osteogenic potential of the CsinCPI-2 protein represents a new insight into plants cysteine proteases inhibitors and this effect needs to be better addressed. The aim of this study was to investigate the performance of pre-osteoblasts in response to CsinCPI-2, mainly focusing on cell adhesion, proliferation and differentiation mechanisms. Together our data show that in the first hours of treatment, protein in CsinCPI-2 promotes an increase in the expression of adhesion markers, which decrease after 24 h, leading to the activation of Kinase-dependent cyclines (CDKs) modulating the transition from G1 to S phases cell cycle. In addition, we saw that the increase in ERK may be associated with activation of the differentiation profile, also observed with an increase in the B-Catenin pathway and an increase in the expression of Runx2 in the group that received the treatment with CsinCPI-2.
植物凝集素是存在于植物中的内源性半胱氨酸蛋白酶抑制剂。它们参与初始发芽率和植物对植物病原体的防御机制。最近，一种从甜橙中提取的新植物凝集素 CsinCPI-2 已被证明能抑制人组织蛋白酶的酶活性，在人牙髓细胞中表现出抗炎潜力和促成骨作用。CsinCPI-2 蛋白的成骨潜力为植物半胱氨酸蛋白酶抑制剂提供了新的见解，需要更好地研究这种作用。本研究旨在研究前成骨细胞对 CsinCPI-2 的反应，主要关注细胞黏附、增殖和分化机制。我们的数据表明，在治疗的最初几个小时内，CsinCPI-2 蛋白促进了黏附标志物的表达增加，24 小时后减少，导致依赖激酶的细胞周期素（CDKs）的激活，调节细胞周期从 G1 期向 S 期的过渡。此外，我们还发现 ERK 的增加可能与分化特征的激活有关，这也与 B-连环蛋白途径的增加和接受 CsinCPI-2 处理的组中 Runx2 表达的增加有关。
