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骨碎补总黄酮抑制NF-kB信号通路干预大鼠椎间盘退变 被引量:1

Rhizoma drynariae flavonoids inhibits rat intervertebral disc degeneration by inhibiting nuclear factor-kappa B signaling pathway
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摘要 背景:椎间盘退变是由于一系列的机械压迫、炎性因子、基质金属蛋白酶等共同作用导致的。骨碎补总黄酮被证实具有抗炎作用,但其延缓椎间盘软骨细胞作用和机制尚不明确。目的:探讨骨碎补总黄酮干预SD大鼠颈椎间盘退变模型的作用机制并对其临床价值进行分析。方法:SD大鼠50只(雌雄各半)随机分为2组:10只作为假手术组,40只大鼠用于建立动静力失衡性颈椎间盘退变大鼠模型。通过病理学方法鉴定造模成功后,取36只颈椎间盘退变模型大鼠随机分为骨碎补总黄酮低、中、高剂量组和模型各9只。假手术组和模型组每日分别给予4 mL生理盐水灌胃,骨碎补总黄酮低、中、高剂量组分别给予62.5,125,250 mg/kg的骨碎补供试液灌胃,各组共喂养1个月,然后分别取出大鼠的C3-4,C4-5,C5-6,C6-7椎间盘组织,采用免疫组织化学、ELISA测定肿瘤坏死因子α、基质金属蛋白酶3蛋白表达;RT-PCR检测肿瘤坏死因子α、基质金属蛋白酶基因表达;Western Blot检测IkBα的磷酸化水平。实验方案经安徽医科大学动物实验伦理委员会批准。结果与结论:①RT-PCR、免疫组织化学、ELISA和Western blot检测显示模型组IkBα的磷酸化水平、肿瘤坏死因子α、基质金属蛋白酶3 mRNA及蛋白表达显著高于假手术组(P<0.01);骨碎补总黄酮干预各组IkBα的磷酸化水平、肿瘤坏死因子α、基质金属蛋白酶3 mRNA及蛋白表达显著低于模型组,以骨碎补总黄酮低剂量组干预效果最佳;②结果说明,骨碎补总黄酮高、中、低剂量均能通过抑制退变椎间盘中NF-kB信号通路激活,以及细胞内基质金属蛋白酶3和肿瘤坏死因子α的表达,保护椎间盘软骨细胞,延缓椎间盘,但以低剂量骨碎补总黄酮干预效果最佳。 BACKGROUND:The combined action of mechanical compression,inflammatory factors,and matrix metalloproteinases leads to intervertebral disc degeneration.Rhizoma drynaria flavones have been shown to have anti-inflammatory effect.However,its protective effects against degeneration of the intervertebral discs remain unclear.OBJECTIVE:To investigate the potential molecular mechanisms underlying the protective effects of rhizoma drynaria flavones in a rat model of cervical intervertebral discs degeneration and analyze the clinical value.METHODS:Fifty Sprague-Dawley rats(half male and half female)were randomly divided into two groups:10 in sham-operated group and 40 in degeneration group.Animal models of cervical intervertebral discs degeneration due to dynamic and static imbalance were made in the degeneration group.After successful modeling,36 model rats were randomized into model group,high-dose group,middle-dose group and low-dose group(n=9 per group).Rats in the sham and model groups were intragastrically given 4 mL of normal saline per day,and those in the high-,middle-and low-dose groups were given 62.5,125,and 250 mg/kg rhizoma drynaria solution,respectively.The C3-4,C4-5,C5-6,and C6-7 segments of the rat spine were taken.Immunohistochemistry and ELISA were used to detect expression of tumor necrosis factorαand matrix metalloproteinase 3 proteins.Quantitative RT-PCR was used to determine the mRNA expression of tumor necrosis factorαand matrix metalloproteinase 3.Western blot was used to detect the phosphorylation of IkBα.RESULTS AND CONCLUSION:Results from the immunohistochemistry,western blot,and RT-PCR,and ELISA showed that the phosphorylation of IkBαand expression of tumor necrosis factorαand matrix metalloproteinase 3 were significantly higher in the model group than the sham group(P<0.01).However,interventions with rhizoma drynaria flavones,especially with low-dose rhizoma drynaria flavones,significantly lowered the phosphorylation of IkBαand expression of tumor necrosis factorαand matrix metalloprotei
作者 陈敏 赵凯 王娟 胡勇 Chen Min;Zhao Kai;Wang Juan;Hu Yong(Department of Rehabilitation,First Affiliated Hospital of Anhui Medical University,Hefei 230000,Anhui Province,China;Department of Orthopedics,First Affiliated Hospital of Anhui Medical University,Hefei 230000,Anhui Province,China)
出处 《中国组织工程研究》 CAS 北大核心 2020年第17期2654-2659,共6页 Chinese Journal of Tissue Engineering Research
基金 安徽省自然科学基金(1708085MH215),项目负责人:胡勇。
关键词 骨碎补总黄酮 椎间盘退变 模型 炎症 基质金属蛋白酶 NF-KB rhizoma drynaria flavones intervertebral disc degeneration model inflammation intracellular matrix metalloproteinase nuclear factor-κB
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