Chinese Journal of Tissue Engineering Research ›› 2021, Vol. 25 ›› Issue (2): 258-263.doi: 10.3969/j.issn.2095-4344.2976
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Yu Chengshuai, Du Gang, Pang Shenning, Lao Shan
Received:
2019-12-19
Revised:
2019-12-24
Accepted:
2020-02-12
Online:
2021-01-18
Published:
2020-11-21
Contact:
Lao Shan, MD, Chief physician, First Affiliated Hospital of Guangxi Medical University, Nanning 530021, Guangxi Zhuang Autonomous Region, China
About author:
Yu Chengshuai, Master candidate, First Affiliated Hospital of Guangxi Medical University, Nanning 530021, Guangxi Zhuang Autonomous Region, China
Supported by:
CLC Number:
Yu Chengshuai, Du Gang, Pang Shenning, Lao Shan. Chemerin, a pro-inflammatory adipokine, regulates chondrocyte proliferation and metabolism by increasing production of nitric oxide[J]. Chinese Journal of Tissue Engineering Research, 2021, 25(2): 258-263.
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2.6 NO的浓度结果 见图6,NO在体内或水溶液中极易氧化成NO2-,在酸性条件下,NO2-与重氮盐磺酸胺生成重氮化合物,进一步与奈基乙烯基二胺偶合,产物在550 nm处有特征性吸收峰,测定其A值,可以计算NO浓度。体外药物刺激2 d后,软骨细胞中NO的生成量较对照组增加,而当Chemerin和白细胞介素1β共存时,NO的生成明显增加(图6A)。这些结果与既往的研究一致,白细胞介素1β可以促进NO的分泌,也表明Chemerin具有类似的作用。当在Chemerin中加入L-NMMA后,NO的分泌明显减少(图6B)。换句话说,L-NMMA成功地抑制了由诱导型一氧化氮合酶合成的NO,表明Chemerin通过促进软骨细胞中诱导型一氧化氮合酶表达而增加NO产生进而影响软骨细胞存活的这一过程可被L-NMMA所阻断。 "
2.7 软骨细胞代谢基因分析 基因的相对表达量见图7。在培养2 d后,通过对软骨细胞蛋白聚糖(一种主要由糖胺聚糖组成的蛋白多糖)、Ⅱ型胶原a1、基质金属蛋白酶13和一氧化氮合酶2基因表达的相对定量检测,进一步研究 Chemerin对软骨细胞细胞外基质合成和降解的影响。如图7A所示,与对照组相比,3个药物实验组细胞中蛋白聚糖和Ⅱ型胶原a1类软骨形成标记基因的相对表达水平均降低,而与细胞外基质降解密切相关的基因一氧化氮合酶2和基质金属蛋白酶13的表达水平显著增加。此外,实验结果表明Chemerin的作用可能不像白细胞介素1β那样强大,但两者之间的协同作用在基因水平上也是显而易见的。在Chemerin组中加入L-NMMA后,基质金属蛋白酶13的相对表达也明显降低(图7B),与图6B的结果一致,这也反过来证明NO能促进基质金属蛋白酶13基因的表达。"
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