摘要
目的探讨身痛逐瘀汤含药血清是否可通过介导核因子-κB(NF-κB)通路抑制肿瘤坏死因子-α(TNF-α)诱导的大鼠髓核细胞退变。方法将成年雄性SD大鼠随机分为4组制备空白血清和身痛逐瘀汤高、中、低剂量含药血清。取从大鼠尾盘获得并传代到第3代的髓核细胞,分为5组培养:空白组加20%空白大鼠血清,模型组加20%空白大鼠血清和20 ng/mL TNF-α,身痛逐瘀汤高剂量组加20%身痛逐瘀汤高剂量含药血清和20 ng/mL TNF-α,身痛逐瘀汤中剂量组加20%身痛逐瘀汤中剂量含药血清和20 ng/mL TNF-α,身痛逐瘀汤低剂量组加20%身痛逐瘀汤低剂量含药血清和20 ng/mL TNF-α,均培养48 h。采用RT-PCR法检测髓核细胞中基质金属蛋白酶-3(MMP-3)、聚集蛋白聚糖(Aggrecan)、Ⅱ型胶原蛋白(CollagenⅡ)、B细胞淋巴瘤/白血病-2(Bcl-2)、Bcl-2家族相关x蛋白(Bax)、半胱氨酸天冬氨酸蛋白酶-3(Caspase-3)mRNA表达情况,采用Western blot法检测细胞中p-IKKα/β、IKKβ、p-IκBa、IκBa、p-p65、p65蛋白表达情况。结果与空白组比较,模型组细胞中MMP-3、Bax、Caspase-3 mRNA相对表达量和p-IKKα/β/IKKβ、p-IκBa/IκBa、p-p65/p65相对表达量均明显升高(P均<0.05),Aggrecan、CollagenⅡ、Bcl-2 mRNA相对表达量均明显降低(P均<0.05);与模型组比较,身痛逐瘀汤各组细胞中MMP-3、Bax、Caspase-3 mRNA相对表达量和p-IKKα/β/IKKβ、p-IκBa/IκBa、p-p65/p65相对表达量均明显降低(P均<0.05),Aggrecan、CollagenⅡ、Bcl-2 mRNA相对表达量均明显升高(P均<0.05),且身痛逐瘀汤各组中各指标呈浓度依赖性递减或递增趋势。结论身痛逐瘀汤含药血清可通过抑制NF-κB通路的激活,减少TNF-α诱导的髓核细胞外基质降解和髓核细胞凋亡,延缓髓核细胞退变。
Objective It is to investigate whether the medicated serum of Shentong Zhuyu Decoction(STZYT)inhibits tumor necrosis factor-α(TNF-α)-induced degeneration of rat myeloid nucleus cells by mediating the nuclear factor-κB(NF-κB)pathway.Methods Adult male SD rats were randomly divided into 4 groups to prepare blank serum and medicated serum containing high,medium and low doses of STZYT.The myeloid nucleus cells obtained from the rat tail disc and passaged to the 3rd generation were taken and divided into 5 groups for culture:the blank group was cultured with 20%serum of blank rats,the model group was cultured with 20%serum of blank rats and 20 ng/mL TNF-α,the STZYT high-dose group was cultured with 20%medicated serum containing high dose of STZYT and 20 ng/mL TNF-α,the STZYT middle-dose group was cultured with medicated serum containing high,medium and low doses of STZYT and 20 ng/mL TNF-α,and the STZYT low-dose group was cultured with medicated serum containing high,medium and low doses of STZYT and 20 ng/mL TNF-α,and all of them were cultured for 48 h.The expressions of matrix metalloproteinase-3(MMP-3),aggregated proteoglycan(Aggrecan),collagen typeⅡ(CollagenⅡ),B-cell lymphoma/leukemia-2(Bcl-2),Bcl-2 family-associated x-protein(Bax),and cysteine aspartate protease-3(Caspase-3)mRNA in the myeloid nucleus cells were detected by RT-PCR,and the expressions of p-IKKα/β,IKKβ,p-IκBa,IκBa,p-p65,p65 protein in the cells were detected by Western blot.Results Compared with the blank group,the relative expressions of MMP-3,Bax,Caspase-3 mRNA and the relative expressions of p-IKKα/β/IKKβ,p-IκBa/IκBa,p-p65/p65 in the cells of the model group were significantly increased(all P<0.05),and the relative expressions of Aggrecan,CollagenⅡ,Bcl-2 mRNA were significantly decreased(all P<0.05).Compared with the model group,the relative expressions of MMP-3,Bax,Caspase-3 mRNA and the relative expressions of p-IKKα/β/IKKβ,p-IκBa/IκBa,p-p65/p65 in the cells of each STZYD group were significantly decreased(all P<0.05),and the relative expressions of Aggrecan,CollagenⅡ,Bcl-2 mRNA were significantly increased(all P<0.05),and the decreasing or increasing trends of these indicators were concentration-dependent in each STZYD group.Conclusion The medicated serum of Shentong Zhuyu Decoction can reduce TNF-α-induced myeloid extracellular matrix degradation and myeloid apoptosis by inhibiting the activation of the NF-κB pathway,thus t slow down myeloid degeneration.
作者
芦冲
栗丽丽
马旭凯
韩东卫
LU Chong;LI Lili;MA Xukai;HAN Dongwei(Shanxi Provincial Integratcd TCM And WM Hospital,Taiyuan 030013,Shanxi,China;Heilongjiang University of Chinese Medicine,Harbin 150040,Heilongjiang,China)
出处
《现代中西医结合杂志》
CAS
2023年第22期3074-3078,3086,共6页
Modern Journal of Integrated Traditional Chinese and Western Medicine
基金
国家自然科学基金资助项目(81904090)
山西中医药大学2022年博士科研启动基金科技兴医创新计划项目(2022-B-03)
山西省卫生健康委科研课题(2023101)。
关键词
腰椎退行性疾病
髓核细胞
身痛逐瘀汤
核因子-κB
lumbar degenerative disc disease
myeloid nucleus
Shentong Zhuyu Decoction
nuclear factor-κB
