Department of Microbiology & Immunology, Pusan National University School of Medicine, Yangsan 50611, Republic of Korea.
Department of Molecular Medicine, University of Texas Health at San Antonio, San Antonio, TX 78229, USA.
J Microbiol Biotechnol. 2022 Aug 28;32(8):1017-1025. doi: 10.4014/jmb.2203.03001. Epub 2022 Jul 8.
Bone homeostasis is regulated by constant remodeling through osteogenesis by osteoblasts and osteolysis by osteoclasts and osteoporosis can be provoked when this balance is broken. Present pharmaceutical treatments for osteoporosis have harmful side effects and thus, our goal was to develop therapeutics from intrisincally safe natural products. Fucoidan is a polysaccharide extracted from many species of brown seaweed, with valuable pharmaceutical activities. To intensify the effect of fucoidan on bone homeostasis, we hydrolyzed fucoidan using AMG, Pectinex and Viscozyme. Of these, fucoidan biotransformed by Pectinex (Fu/Pec) powerfully inhibited the induction of tartrate-resistant acid phosphatase (TRAP) activity in osteoclasts differentiated from bone marrow macrophages (BMMs) by the receptor for activation of nuclear factor-κB ligand (RANKL). To investigate potential of lower molecular weight fucoidan it was separated into >300 kDa, 50-300 kDa, and <50 kDa Fu/Pec fractions by ultrafiltration system. The effects of these fractions on TRAP and alkaline phosphatase (ALP) activities were then examined in differentiated osteoclasts and MC3T3-E1 osteoblasts, respectively. Interestingly, 50-300 kDa Fu/Pec suppressed RANKL-induced osteoclasts differentiation from BMMs but did not synergistically enhance osteoblasts differentiation induced by osteogenic agents. In addition, this fraction inhibited the expressions of NFATc1, TRAP, OSCAR, and RANK, which are all key transcriptional factors involved in osteoclast differentiation, and those of Src, c-Fos and Mitf, as determined by RT-PCR. In conclusion, enzymatically low-molecularized 50-300 kDa Fu/Pec suppressed TRAP by downregulating RANKL-related signaling, contributing to the inhibition of osteoclasts differentiation, and represented a potential means of inducing bone remodeling in the background of osteoporosis.
骨稳态是通过成骨细胞的成骨作用和破骨细胞的骨吸收来调节的,当这种平衡被打破时,就会引发骨质疏松症。目前用于治疗骨质疏松症的药物有副作用,因此,我们的目标是从固有安全的天然产物中开发治疗方法。褐藻糖胶是一种从多种褐藻中提取的多糖,具有有价值的药物活性。为了增强褐藻糖胶对骨稳态的作用,我们使用 AMG、pectinex 和 Viscozyme 对褐藻糖胶进行了水解。其中,pectinex 转化的褐藻糖胶(Fu/Pec)有力地抑制了核因子-κB 配体(RANKL)诱导的破骨细胞分化的抗酒石酸酸性磷酸酶(TRAP)活性。为了研究低分子量褐藻糖胶的潜力,它通过超滤系统被分离成>300 kDa、50-300 kDa 和<50 kDa 的 Fu/Pec 级分。然后,在分化的破骨细胞和 MC3T3-E1 成骨细胞中分别检测这些级分对 TRAP 和碱性磷酸酶(ALP)活性的影响。有趣的是,50-300 kDa 的 Fu/Pec 抑制了 RANKL 诱导的 BMMs 来源的破骨细胞分化,但不能协同增强成骨剂诱导的成骨细胞分化。此外,该级分通过 RT-PCR 抑制了 NFATc1、TRAP、OSCAR 和 RANK 的表达,这些都是参与破骨细胞分化的关键转录因子,以及 Src、c-Fos 和 Mitf 的表达。总之,酶解低分子量 50-300 kDa 的 Fu/Pec 通过下调 RANKL 相关信号来抑制 TRAP,有助于抑制破骨细胞分化,并代表了在骨质疏松症背景下诱导骨重塑的一种潜在手段。