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地骨皮对RAW 264.7细胞中RANKL诱导的破骨细胞分化和激活的影响。

The effects of Lycii Radicis Cortex on RANKL-induced osteoclast differentiation and activation in RAW 264.7 cells.

作者信息

Kim Jae-Hyun, Kim Eun-Young, Lee Bina, Min Ju-Hee, Song Dea-Uk, Lim Jeong-Min, Eom Ji Whan, Yeom Mijung, Jung Hyuk-Sang, Sohn Youngjoo

机构信息

Department of Anatomy, College of Korean Medicine, Kyung Hee University, Seoul 130-701, Republic of Korea.

Acupuncture and Meridian Science Research Center, College of Korean Medicine, Kyung Hee University, Seoul 130-701, Republic of Korea.

出版信息

Int J Mol Med. 2016 Mar;37(3):649-58. doi: 10.3892/ijmm.2016.2477. Epub 2016 Feb 1.

Abstract

Post-menopausal osteoporosis is a serious age-related disease. After the menopause, estrogen deficiency is common, and excessive osteoclast activity causes osteoporosis. Osteoclasts are multinucleated cells generated from the differentiation of monocyte/macrophage precursor cells such as RAW 264.7 cells. The water extract of Lycii Radicis Cortex (LRC) is made from the dried root bark of Lycium chinense Mill. and is termed 'Jigolpi' in Korea. Its effects on osteoclastogenesis and post‑menopausal osteoporosis had not previously been tested. In the present study, the effect of LRC on receptor activator of nuclear factor-κB (NF-κB) ligand (RANKL)-induced osteoclast differentiation was demonstrated using a tartrate-resistant acid phosphatase (TRAP) assay and pit formation assay. Moreover, in order to analyze molecular mechanisms, we studied osteoclastogenesis-related markers such as nuclear factor of activated T-cells cytoplasmic 1 (NFATc1), c-Fos, receptor activator of NF-κB (RANK), TRAP, cathepsin K (CTK), matrix metallopeptidase-9 (MMP-9), calcitonin receptor (CTR) and carbonic anhydrase Ⅱ (CAII) using RT-qPCR and western blot analysis. Additionally, we also determined the effect of LRC on an ovariectomized (OVX) rat model. We noted that LRC inhibited RANKL-induced osteoclast differentiation via suppressing osteoclastogenesis-related markers. It also inhibited osteoporosis in the OVX rat model by decreasing loss of bone density and trabecular area. These results suggest that LRC exerts a positive effect on menopausal osteoporosis.

摘要

绝经后骨质疏松症是一种严重的与年龄相关的疾病。绝经后,雌激素缺乏很常见,破骨细胞活性过高会导致骨质疏松症。破骨细胞是由单核细胞/巨噬细胞前体细胞(如RAW 264.7细胞)分化产生的多核细胞。地骨皮水提取物由枸杞干燥根皮制成,在韩国被称为“鸡骨皮”。其对破骨细胞生成和绝经后骨质疏松症的影响此前尚未得到测试。在本研究中,使用抗酒石酸酸性磷酸酶(TRAP)测定法和凹坑形成测定法证明了地骨皮对核因子κB(NF-κB)配体(RANKL)诱导的破骨细胞分化的影响。此外,为了分析分子机制,我们使用RT-qPCR和蛋白质印迹分析研究了破骨细胞生成相关标志物,如活化T细胞核因子细胞质1(NFATc1)、c-Fos、NF-κB受体激活剂(RANK)、TRAP、组织蛋白酶K(CTK)、基质金属蛋白酶-9(MMP-9)、降钙素受体(CTR)和碳酸酐酶Ⅱ(CAII)。此外,我们还确定了地骨皮对去卵巢(OVX)大鼠模型的影响。我们注意到,地骨皮通过抑制破骨细胞生成相关标志物来抑制RANKL诱导的破骨细胞分化。它还通过减少骨密度和小梁面积的损失来抑制OVX大鼠模型中的骨质疏松症。这些结果表明,地骨皮对绝经后骨质疏松症具有积极作用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/56e4/4771095/3929fd297bca/IJMM-37-03-0649-g00.jpg

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