Pan Xin, Xu Kang, Qiu Xuefeng, Zhao Wen, Wang Dong, Yang Li
College of Pharmacy, South-Central University for Nationalities, Wuhan, China.
National Innovation and Attracting Talents "111" Base, Key Laboratory of Biorheological Science and Technology, Ministry of Education, College of Bioengineering, Chongqing University, Chongqing, China; Department of Bioengineering, University of California at Los Angeles, 410 Westwood Plaza, 5121 Engineering V, Los Angeles, CA 90095, USA.
Evid Based Complement Alternat Med. 2016;2016:2057631. doi: 10.1155/2016/2057631. Epub 2016 Nov 23.
This study aimed to investigate the extract components of FP on rat chondrocyte function and cartilaginous formation . Petroleum ether extract (P-e) of FP extract components was selected to treat Sprague-Dawley rat chondrocytes. Cell viability was tested with different concentrations (0.1, 1, 10, and 100 g/mL) of P-e treatment. Concentrations of 0.1 and 1 g/mL P-e conditioned culture mediums were used for treating chondrocytes in experiments. Cell proliferation was measured via DNA incorporation assay. Type II collagen, aggrecan, and Sox-9 genes expression levels were measured with RT-PCR. Additionally, cartilaginous formation was analyzed with type II collagen immunofluorescence, H&E, and alcian blue staining. Concentrations of 0.1 and 1 g/mL P-e showed low cytotoxicity and demonstrated stimulatory effects on chondrocyte proliferation in early stages. Following 6 days of P-e culture, aggrecan and Sox-9 gene expression levels of the 1 g/mL P-e group were upregulated by 1.82- ( < 0.05) and 2.06-fold ( < 0.05), respectively, versus controls. Moreover, 1 g/mL P-e significantly stimulated cell aggregation and type II collagen deposits after 1 week of treatment. Noteworthy, tight cartilaginous structures formed in the 10-day 1 g/mL P-e conditioned culture. These findings suggest that P-e has the potential to treat cartilage degeneration induced by chondrocyte failure.
本研究旨在探讨FP的提取物成分对大鼠软骨细胞功能和软骨形成的影响。选取FP提取物成分的石油醚提取物（P-e）处理Sprague-Dawley大鼠软骨细胞。用不同浓度（0.1、1、10和100 g/mL）的P-e处理来检测细胞活力。在实验中，使用浓度为0.1和1 g/mL的P-e条件培养基处理软骨细胞。通过DNA掺入法测定细胞增殖。用RT-PCR检测II型胶原蛋白、聚集蛋白聚糖和Sox-9基因的表达水平。此外，用II型胶原蛋白免疫荧光、苏木精-伊红（H&E）和阿尔辛蓝染色分析软骨形成。浓度为0.1和1 g/mL的P-e显示出低细胞毒性，并在早期对软骨细胞增殖具有刺激作用。P-e培养6天后，1 g/mL P-e组的聚集蛋白聚糖和Sox-9基因表达水平分别比对照组上调了1.82倍（<0.05）和2.06倍（<0.05）。此外，处理1周后，1 g/mL P-e显著刺激细胞聚集和II型胶原蛋白沉积。值得注意的是，在10天的1 g/mL P-e条件培养中形成了紧密的软骨结构。这些发现表明，P-e有潜力治疗由软骨细胞功能衰竭引起的软骨退变。
