| 过载静压下益肾活血通络含药血清对人椎间盘髓核细胞凋亡的影响 |
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Received:September 16, 2020
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| 作者 | Author | 单位 | Unit | E-Mail |
| 肖辉灯 |
XIAO Hui-deng |
首都医科大学附属北京中医医院, 北京 100010 |
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| 柳根哲 |
LIU Gen-zhe |
首都医科大学附属北京中医医院, 北京 100010 |
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| 孙旗 |
SUN Qi |
北京中医药大学东直门医院, 北京 100700 |
Dongzhimen Hospital, Beijing University of Chinese Medicine, Beijing 100700, China |
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| 张帆 |
ZHANG Fan |
北京中医药大学东直门医院, 北京 100700 |
Dongzhimen Hospital, Beijing University of Chinese Medicine, Beijing 100700, China |
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| 祝永刚 |
ZHU Yong-gang |
首都医科大学附属北京中医医院, 北京 100010 |
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| 刘志超 |
LIU Zhi-chao |
首都医科大学附属北京中医医院, 北京 100010 |
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| 郭菲宇 |
GUO Fei-yu |
首都医科大学附属北京中医医院, 北京 100010 |
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| 康晓乐 |
KANG Xiao-le |
北京中医药大学东直门医院, 北京 100700 |
Dongzhimen Hospital, Beijing University of Chinese Medicine, Beijing 100700, China |
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| 袁巧妹 |
YUAN Qiao-mei |
北京中医药大学东直门医院, 北京 100700 |
Dongzhimen Hospital, Beijing University of Chinese Medicine, Beijing 100700, China |
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| 邸学士 |
DI Xue-shi |
北京中医药大学东直门医院, 北京 100700 |
Dongzhimen Hospital, Beijing University of Chinese Medicine, Beijing 100700, China |
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| 龙水文 |
LONG Shui-wen |
北京中医药大学东直门医院, 北京 100700 |
Dongzhimen Hospital, Beijing University of Chinese Medicine, Beijing 100700, China |
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| 陈江 |
CHEN Jiang |
北京中医药大学东直门医院, 北京 100700 |
Dongzhimen Hospital, Beijing University of Chinese Medicine, Beijing 100700, China |
42691631@qq.com |
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| 期刊信息：《中国骨伤》2021年34卷，第11期，第1044-1052页 |
| DOI：10.12200/j.issn.1003-0034.2021.11.012 |
| 基金项目:国家自然科学基金委员会青年科学基金项目（编号：81603638）；中国博士后科学基金面上项目（编号：2019M662791）；中央高校基本科研业务费专项资金资助（编号：2018-JYB-XJQ010）；北京中医药大学2019年度基本科研项目（编号：2019-JYB-JS-042）；北京中医药大学东直门医院青苗人才项目（编号：DZMYS-201702） |
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目的：探讨在过载静压应力下益肾活血通络方含药血清对人椎间盘髓核细胞凋亡的影响及其相关机制。
方法：将人椎间盘髓核细胞分为3组，空白组无任何干预，模型组和中药血清干预组在体外3 MPa压应力干预下作用2、4、6 h后，观察椎间盘髓核细胞的形态、生长状况、超微结构的变化及差异；检测椎间盘髓核细胞的凋亡率及髓核细胞内核因子κB p65（nuclear factor kappa-B p65，NF-κB p65），Y染色体中性别决定区相关的高迁移率组框9（SRY-related high mobility group-box 9，SOX9），C/EBP同源蛋白（C/EBP-homologous protein，CHOP），基质金属蛋白酶-13（matrix metalloprotein-13，MMP-13）蛋白和相对应基因表达情况。
结果：同一作用时间下模型组髓核细胞与空白组比较：体积缩小、胞浆减少、生长状况更差；中药血清干预组髓核细胞较模型组体积稍大，形态保存更完整，胞浆更丰富，生长状况更好。同一作用时间下空白组髓核细胞超微结构完整，主级突起、次级突起结构未见断裂，模型组与中药血清干预组超微结构破坏，主级突起、次级突起可见不同程度断裂，两组超微结构未见明显差异。相同作用时间下模型组髓核细胞凋亡率高于空白组，而中药血清干预组髓核细胞凋亡率低于模型组（P<0.05）；随作用时间增加，空白组、中药血清干预组椎间盘髓核细胞凋亡率未见明显差异，模型组椎间盘髓核细胞凋亡率增加。相同作用时间下，中药血清干预组较模型组NF-κB p65、CHOP、MMP-13表达减少，SOX9表达增多（P<0.05）；随作用时间的增加，空白组、模型组髓核细胞的NF-κB p65、CHOP、MMP-13表达增多，SOX9表达减少，且模型组表达程度较空白组大（P<0.05）；通过基因表达总体来看，在相同作用时间下，中药血清干预组髓核细胞基因NF-κB p65、CHOP、MMP-13相对定量较模型组均减少，而SOX9相对定量增多（P<0.05）；空白组髓核细胞基因NF-κB p65、CHOP、MMP-13相对定量比模型组减少，而SOX9相对定量增多（P<0.05）；随作用时间的增加，空白组、模型组髓核细胞的NF-κB p65、CHOP、MMP-13相对定量增多，SOX9相对定量减少（P<0.05）。
结论：益肾活血通络方能减少过载静压下髓核细胞的凋亡，具有延缓髓核细胞退变的作用，其机制可能通过抑制髓核细胞NF-κB p65信号通路，使CHOP、MMP-13表达减少，SOX9表达增加有关。 |
| [关键词]:中草药 髓核 细胞 过载静压压力 细胞凋亡 |
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| Efficacy of Yishen Huoxue Tongluo(益肾活血通络) medicated serum on apoptosis of nucleus pulposus cells of human intervertebral disc under overload static pressure |
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Abstract:
Objective: To explore the efficacy of Yishen Huoxue Tongluo(益肾活血通络) decoction containing serum on the apoptosis of human disc nucleus pulposus cells under the overload static pressure stress and its related mechanism.
Methods: Human nucleus pulposus cells were divided into three groups. The blank group had no intervention. The model group and traditional Chinese medicine serum intervention group were treated with 3 MPa compressive stress in vitro for 2, 4 and 6 hours. The changes and differences of morphology, growth status and ultrastructure of intervertebral disc nucleus pulposus cells were observed. The apoptosis rate of nucleus pulposus cells and nuclear factor kappa-B p65(NF-κB p65), SRY-related high mobility group-box 9(SOX9), C/EBP-homologous protein (CHOP), matrix metalloprotein-13(MMP-13) and corresponding gene expression were detected.
Results: At the same time, compared with the blank group, the nucleus pulposus cells in the model group were smaller in volume, less in cytoplasm and worse in growth; the nucleus pulposus cells in the traditional Chinese medicine serum intervention group were slightly larger in volume, more complete in morphology, richer in cytoplasm and better in growth. Under the same action time, the ultrastructure of nucleus pulposus cells in blank group was complete, and the structures of primary and secondary processes were not broken;and the ultrastructure of model group and traditional Chinese medicine serum intervention group were damaged, the main and secondary processes were broken to varying degrees, and there was no significant difference between the two groups. At the same time, the apoptosis rate of nucleus pulposus cells in model group was higher than that in blank group, while the apoptosis rate of nucleus pulposus cells in the traditional Chinese medicine serum intervention group was lower than that in model group, the difference was statistically significant(P<0.05);with the increase of action time, there was no significant difference in the apoptosis rate of nucleus pulposus cells between blank group and traditional Chinese medicine serum intervention group, and the apoptosis rate of nucleus pulposus cells in model group was increased. Compared with model group, the expression of NF-κB p65, CHOP, MMP-13 were decreased and SOX9 was increased in traditional Chinese medicine serum intervention group at the same time(P<0.05);with the increase of action time, the expression of NF-κB p65, CHOP and MMP-13 were increased, and the expression of SOX9 was decreased in blank group and model group(P<0.05), and the expression level of model group was higher than that of blank group(P<0.05). Overall observation by gene expression, under the same action time, the relative quantifications of NF-κB p65, CHOP and MMP-13 in traditional Chinese medicine serum intervention group were lower than that in model group, while SOX9 was increased(P<0.05);compared with model group, the relative quantifications of NF-κB p65, CHOP and MMP-13 in blank group were decreased(P<0.05), and the relative quantification of SOX9 was increased(P<0.05);with the increase of action time, the relative quantifications of NF-κB p65, CHOP and MMP-13 of nucleus pulposus cells in blank group and model group were increased and SOX9 was decreased(P<0.05).
Conclusion: Yishen Huoxue Tongluo(益肾活血通络) decoction can reduce the apoptosis of nucleus pulposus cells under overload and static pressure, and has the effect of delaying the degeneration of nucleus pulposus cells. Its mechanism may be related to the decrease of CHOP, MMP-13 expression and the increase of SOX9 expression by inhibiting NF-κB p65 signal pathway of nucleus pulposus cells. |
| KEYWORDS:Drugs,Chinese herbal Nucleus pulposus Cells Overload static pressure Apoptosis |
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| 引用本文，请按以下格式著录参考文献： |
| 中文格式: | 肖辉灯,柳根哲,孙旗,张帆,祝永刚,刘志超,郭菲宇,康晓乐,袁巧妹,邸学士,龙水文,陈江.过载静压下益肾活血通络含药血清对人椎间盘髓核细胞凋亡的影响[J].中国骨伤,2021,34(11):1044~1052 |
| 英文格式: | XIAO Hui-deng,LIU Gen-zhe,SUN Qi,ZHANG Fan,ZHU Yong-gang,LIU Zhi-chao,GUO Fei-yu,KANG Xiao-le,YUAN Qiao-mei,DI Xue-shi,LONG Shui-wen,CHEN Jiang.Efficacy of Yishen Huoxue Tongluo(益肾活血通络) medicated serum on apoptosis of nucleus pulposus cells of human intervertebral disc under overload static pressure[J].zhongguo gu shang / China J Orthop Trauma ,2021,34(11):1044~1052 |
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