| 摘要: |
| 目的：观察低氧诱导因子-1α小干扰质粒（small interfering hypoxia-inducible factor-1α，siHIF-1α）干预前后补肾活血汤对大鼠椎间盘退变髓核细胞炎症因子及细胞增殖的影响。方法：分离培养原代大鼠椎间盘髓核细胞，采用免疫荧光技术对第2代细胞进行鉴定；取第3代大鼠髓核细胞，采用白细胞介素-1β（interleukin-1β，IL-1β）进行退变髓核细胞造模，将造模成功的大鼠髓核细胞随机分为5组：空白组，模型组，和补肾活血汤低、中、高剂量组，并给予不同浓度药剂干预。采用细胞活力-毒性检测试验盒8（cell counting kit-8，CCK-8）检测各组细胞增殖情况，流式细胞术检测各组细胞凋亡率，生化分析法检测细胞氧化应激因子超氧化物歧化酶（superoxide dismutase，SOD）、丙二醛（malondialdethde，MDA）水平；酶联免疫吸附法检测炎症因子白细胞介素-6（interleukin-6，IL-6）的表达水平。结果：本研究成功分离、培养得到大鼠髓核细胞；相比空白组，模型组大鼠髓核细胞增殖减弱，凋亡率增加；补肾活血汤各剂量组能抑制髓核细胞凋亡率（P＜0.05）、抑制 IL-1β对氧化应激因子SOD的下调作用（P＜0.05）及对 MDA含量的上调作用（P＜0.05）；补肾活血汤各剂量组中胱天蛋白酶-3（cysteine aspartic acid specific protease-3，caspase-3）、胱天蛋白酶-9（cysteine aspartic acid specific protease-9，caspase-9）的表达水平较模型组均明显下调（P＜0.05）；与空白组相比，补肾活血汤各剂量组低氧诱导因子-1α（hypoxia-inducible factor-1α，HIF-1α）升高，与模型组相比，补肾活血汤各剂量组HIF-1α表达升高，当加入siHIF-1α后，大鼠髓核细胞增殖减弱，凋亡率增加，SOD下调，MDA、IL-1β、IL-6含量上调。结论：HIF-1α对髓核细胞的增殖呈正相关，并且能抑制炎症因子表达，补肾活血汤可能通过调控HIF-1α实现抑制大鼠椎间盘退变模型髓核细胞凋亡、炎症和氧化应激的发生，以补肾活血汤中剂量组为佳。 |
| 关键词: 低氧诱导因子-1α 补肾活血汤 大鼠髓核细胞 增殖与凋亡 炎症因子 |
| DOI： |
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| Effect of Bushen Huoxue decoction on the proliferation of nucleus pulposus cells and inflammatory factors in rat intervertebral disc before and after intervention with small interfering hypoxia-inducible factor-1α |
| CHEN Haoxiong,JIN Daxiang,XU Qiliang |
| (Shenzhen Hospital of Guangzhou University of Chinese Medicine（Fu Tian），Shenzhen 518034，China;The First Affiliated Hospital of Guangzhou University of Chinese Medicine，Guangzhou 510405，Guangdong，China) |
| Abstract: |
| Objective：To investigate the effect of Bushen Huoxue decoction on inflammatory factors and proliferation of nucleus pulposus cells in rats with intervertebral disc degeneration before and after intervention with small interfering hypoxia-inducible factor-1α （siHIF-1α）.Methods：Primary nucleus pulposus cells of the rat intervertebral disc were separated and culture，and immunofluorescence assay was used for identification of the second passage of cells.The third passage of cells were collected，and interleukin-1β （IL-1β）was used to establish a nucleus pulposus cell model of degeneration.After successful modeling，the nucleus pulposus cells were randomly divided into blank group，model group，and low-，middle-，and high-dose Bushen Huoxue decoction groups，and medi-cament at different concentrations was used for intervention.CCK-8 assay was used to measure cell proliferation in each group；flow cytometry was used to measure cell apoptosis rate；biochemical methods were used to measure the levels of the oxidative stress factors superoxide dismutase （SOD）and malondialdehyde （MDA）in cells；ELISA was used to measure the expression level of the inflammatory factor interleukin-6 （IL-6）.Results：Rat nucleus pulposus cells were successfully separated and cultured in this study.Compared with the blank group，the model group had a reduction in cell proliferation and an increase in apoptosis rate.Bushen Huoxue decoction could inhibit the apoptosis rate of nucleus pulposus cells （P＜0.05）and suppress IL-1β to downregulate the oxidative stress factor SOD （P＜0.05）and upregulate the content of MDA （P＜0.05）.Compared with the model group，the Bushen Huoxue decoction groups had significant reductions in the expression levels of caspase-3 and caspase-9 （P＜0.05）；compared with the blank group，the Bushen Huoxue decoction groups had an increase in hypoxia-inducible factor 1α （HIF-1α），and compared with the model group，each Bushen Huoxue decoction group had an increase in the expression of HIF-1α.After the addition of siHIF-1α，the rat nucleus pulposus cells showed a reduction in proliferation，an increase in apoptosis rate，a reduction in SOD，and increases in MDA，IL-1β，and IL-6.Conclusion：HIF-1α is positively correlated with the proliferation of nucleus pulposus cells and can inhibit the expression of inflammatory factors.Bushen Huoxue decoction can inhibit apoptosis，inflammation，and oxidative stress in nucleus pulposus cells as a model of intervertebral disc degeneration in rats，possibly by regulating HIF-1α，and middle-dose Bushen Huoxue decoction has the best effect. |
| Key words: hypoxia-inducible factor 1α Bushen Huoxue decoction rat nucleus pulposus cells proliferation and apoptosis inflammatory factor |
