| 引用本文:刘志超1,2，张帆3，孙旗3，康晓乐3，袁巧妹3，柳根哲4，陈江3,5.身痛逐瘀汤对静水压下人髓核细胞凋亡和基质相关蛋白表达的影响[J].世界中医药,2020,(08):. |
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| 身痛逐瘀汤对静水压下人髓核细胞凋亡和基质相关蛋白表达的影响 |
| Effects of Shentong Zhuyu Decoction on Autophagy,Matrix Related Genes and Expression of Sox9 and Collagen Ⅱ in Human Nucleus Pulposus Cells under Hydrostatic Pressure |
| 投稿时间：2019-08-17 |
| DOI：10.3969/j.issn.1673-7202.2020.08.004 |
| 中文关键词: 椎间盘退变 身痛逐瘀汤 人髓核细胞 静水压 凋亡 基质代谢 |
| English Keywords:PI3K/AKT pathway Shentong Zhuyu decoction Human nucleus pulposus cells Hydrostatic pressure Apoptosis Matrix metabolism |
| 基金项目:国家自然科学基金项目(81603638);中国博士后科学基金面上项目(2019M662791);中央高校基本科研业务费专项资金资助(2018-JYB-XJQ010);北京中医药大学2017年度基本科研业务费项目(2017-JYB-JS-085);北京中医药大学2019年度基本科研业务费项目(2019-JYB-JS-042);北京中医大学东直门医院青苗人才项目(DZMYS-201702) |
| 作者 | 单位 | | 刘志超1,2，张帆3，孙旗3，康晓乐3，袁巧妹3，柳根哲4，陈江3,5 | 1 中国中医科学院广安门医院南区骨一科,北京,102600
2 北京中医药大学,北京,100029
3 北京中医药大学东直门医院骨伤一科,北京,100700
4 首都医科大学附属北京中医医院骨伤科,北京,100010
5 湖南中医药大学,长沙,410208 |
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| 目的:探讨身痛逐瘀汤对静水压下人髓核细胞(NP)凋亡率及髓核细胞外基质相关蛋白SOX9、Collagen Ⅱ表达的调控作用。方法：按照兔与人的表面积转换方法得出兔中药灌胃量为28 g/(kg·d)颗粒剂。连续灌胃3 d后,从腹主动脉取血,制备身痛逐瘀汤含药血清。将人NP细胞分为6组,压力组:0.3 MPa压力组、1 MPa压力组、3 MPa压力组;中药组:0.3 MPa压力+中药含药血清组、1 MPa压力+中药含药血清组、3 MPa压力+中药含药血清组。将6组人NP细胞在医用恒温静水压压力罐中作用2 h、4 h、6 h后,使用Cell Counting Kit-8（CCK-8）法检测每组NP细胞的增殖活性；使用Annexin V-FITC/Propidium Iodide双染法检验各组NP细胞的凋亡情况;Western Blot法检验SOX9、Collagen Ⅱ在NP细胞中含量的变化。结果：CCK-8法检验2组NP细胞的活性:在相同压力和时间下,含药血清组NP细胞活性比压力组高,差异有统计学意义（P<0.05）；流式细胞术检测NP细胞凋亡:在相同压力和时间下,含药血清组NP细胞凋亡率低于压力组,差异有统计学意义（P<0.05）。Western Blot检测相关蛋白的表达情况:中药组SOX9、Collagen II的总体水平高于压力组,差异有统计学意义（P<0.05）。结论：在静水压环境下下,身痛逐瘀汤能够增加细胞活性,减少细胞凋亡,并促进人NP细胞 Collagen Ⅱ、SOX9表达,具有延缓椎间盘退变的作用。 |
| English Summary: |
| To investigate the regulation of Shentong Zhuyu Decoction on apoptotic rate of human nucleus pulposus(NP) cells and expression of anti-apoptotic proteins SOX9 and Collagen II under hydrostatic pressure.Methods:According to the conversion method of “rabbit surface area/human surface area=rabbit dosage/human dosage”,the dosage of 28 g/kg D granules for rabbits was calculated.After 3 days of continuous intragastric administration,abdominal aorta blood was taken to prepare serum containing Shentong Zhuyu Decoction.Human NP cells were divided into 6 groups:pressure group:0.3 MPa pressure group,1 MPa pressure group,3 MPa pressure group; Chinese medicine group:0.3 MPa pressure+Chinese medicine serum group,1 MPa pressure+Chinese medicine serum group,3 MPa pressure+Chinese medicine serum group.Cell Counting Kit-8 (CCK-8) method was used to detect the proliferative activity of NP cells in 6 groups of human NP cells after 2,4 and 6 hours in a constant temperature hydrostatic pressure tank.Annexin V-FITC/Propidium Iodide double staining flow cytometry was used to detect the apoptotic status of NP cells in each group.Changes of SOX9 and Collagen II content in NP cells were detected using Western Blot.Results:CCK-8 method was used to test the activity of NP cells in the two groups.The activity of NP cells in the drug-containing serum group was higher than that in the pressure group at the same pressure and time (P<0.05).Flow cytometry was used to test the apoptosis of NP cells.The apoptosis rate of NP cells in the drug-containing serum group was lower than that in the pressure group at the same pressure and time (P<0.05).Western Blot was used to detect the expression of related proteins:SOX9 and Collagen II levels in the Chinese herbal medicine group were higher (P<0.05).Conclusion:Shentong Zhuyu Decoction can activate PI3K/AKT pathway, promote the expression of Collagen II and SOX9,increase the activity of NP cells, reduce autophagy and apoptosis of NP cells,thus delaying the degeneration of intervertebral disc. |
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