Li Nianhu, Whitaker Camden, Xu Zhanwang, Heggeness Michael, Yang Shang-You
Department of Orthopedics, University of Kansas School of Medicine-Wichita, 929 N Saint Francis St, Wichita, KS, USA; Department of Orthopaedic Surgery, Affiliated Hospital to Shandong University of Traditional Chinese Medicine, 42 Wenhua West Road, Jinan, China.
Department of Orthopedics, University of Kansas School of Medicine-Wichita, 929 N Saint Francis St, Wichita, KS, USA.
Spine J. 2016 Oct;16(10):1231-1237. doi: 10.1016/j.spinee.2016.05.007. Epub 2016 May 18.
Over half the population of the world will suffer from moderate or severe low back pain (LBP) during their life span. Studies have shown that naringin, a major flavonoid in grapefruit and an active compound extracted from a Chinese herbal medicine (Rhizoma Drynariae) possesses many pharmacological effects.
The aim of this study was to evaluate the influence of naringin on the growth of degenerative human nucleus pulposus (NP) cells, and its repair effects on protein and gene expressions of the cells.
STUDY DESIGN/SETTING: This was an in vitro investigation of the human NP cells isolated from degenerated intervertebral discs that were interacted with various concentrated of naringin.
This study was exempted by the institutional Human Subjects Committee-2, University of Kansas School of Medicine-Wichita. Degenerative human NP cells were isolated from intervertebral discs of patients with discogenic LBP and cultured at 37°C with 5% CO. The proliferation of NP cells was determined following treatment with various concentrations of naringin. The protein expressions of tumor necrosis factor-α (TNF-α) and Bone morphogenetic protein 2 (BMP-2) were tested using enzyme-linked immunosorbent assay. Aggrecan and type II collagen levels were measured by immunohistological staining. Further examination of the gene expression of aggrecan, Sox6, and MMP3 was performed after intervention with naringin for 3 days.
The human NP cells were successfully propagated in culture and stained positive with toluidine blue staining. Naringin effectively enhanced the cell proliferation at an optimal concentration of 20 µg/mL. Naringin treatment resulted in significant inhibition of TNF-α, but elevated protein expressions of BMP-2, collagen II, and aggrecan. Naringin also increased disc matrix gene activity including aggrecan and Sox6, and decreased the gene expression of MMP3.
Naringin effectively promotes the proliferation of degenerative human NP cells and improves the recuperation of the cells from degeneration by increasing expression of aggrecan, BMP-2, and Sox6 while inhibiting the expression of TNF-α and MMP3. This study suggests that naringin may represent an alternative therapeutic agent for disc degeneration.
世界上超过一半的人口在其一生中会遭受中度或重度腰痛(LBP)。研究表明,柚皮苷是葡萄柚中的一种主要黄酮类化合物,也是从一种中药材(骨碎补)中提取的活性化合物,具有多种药理作用。
本研究旨在评估柚皮苷对退变人髓核(NP)细胞生长的影响及其对细胞蛋白质和基因表达的修复作用。
研究设计/环境:这是一项对从退变椎间盘分离的人NP细胞与不同浓度柚皮苷相互作用的体外研究。
本研究获堪萨斯大学医学院威奇托分校机构人类受试者委员会2批准豁免。从椎间盘源性LBP患者的椎间盘中分离退变的人NP细胞,并在37°C、5%CO₂条件下培养。用不同浓度的柚皮苷处理后测定NP细胞的增殖情况。采用酶联免疫吸附测定法检测肿瘤坏死因子-α(TNF-α)和骨形态发生蛋白2(BMP-2)的蛋白表达。通过免疫组织化学染色测量聚集蛋白聚糖和II型胶原蛋白水平。用柚皮苷干预3天后进一步检测聚集蛋白聚糖、Sox6和MMP3的基因表达。
人NP细胞在培养中成功增殖,甲苯胺蓝染色呈阳性。柚皮苷在最佳浓度20μg/mL时有效促进细胞增殖。柚皮苷处理导致TNF-α显著抑制,但BMP-2、胶原蛋白II和聚集蛋白聚糖的蛋白表达升高。柚皮苷还增加了包括聚集蛋白聚糖和Sox6在内的椎间盘基质基因活性,并降低了MMP3的基因表达。
柚皮苷通过增加聚集蛋白聚糖、BMP-2和Sox6的表达,同时抑制TNF-α和MMP3的表达,有效促进退变人NP细胞的增殖,并改善细胞从退变中的恢复。本研究表明柚皮苷可能是椎间盘退变的一种替代治疗药物。