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李硕夫,刘宏哲,张嘉麟,杨雷,陈龙,郭彦涛,刘笑蓉.牛膝总皂苷对IL-1β诱导髓核细胞的凋亡及炎性损伤的影响[J].湖南中医药大学学报英文版,2023,43(9):1591-1597.[Click to copy
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| 牛膝总皂苷对IL-1β诱导髓核细胞的凋亡及炎性损伤的影响 |
| 李硕夫,刘宏哲,张嘉麟,杨雷,陈龙,郭彦涛,刘笑蓉 |
| (湖南中医药大学第一附属医院, 湖南 长沙 410007;湖南省人民医院, 湖南 长沙 410002;中南大学, 湖南 长沙 410083;湖南中医药大学, 湖南 长沙 410208) |
| 摘要: |
| 目的 通过观察髓核细胞（nucleus pulposus cell, NPC）的凋亡、炎症及细胞外基质（extracellular matrix, ECM）代谢研究不同浓度牛膝总皂苷（achyranthes bidentata saponin, ABS）对白细胞介素-1β（interleukin-1β, IL-1β）诱导人源NPC损伤的保护作用。方法 通过IL-1β诱导建立NPC退变模型，随机将细胞分为正常组、模型组（10 ng/mL IL-1β）、ABS低剂量组（10 ng/mL IL-1β+3 μg/mL ABS）、ABS中剂量组（10 ng/mL IL-1β+10 μg/mL ABS）及ABS高剂量组（10 ng/mL IL-1β+30 μg/mL ABS），干预24 h。CCK-8法检测细胞活力，采用Tunel法和流式细胞术检测细胞凋亡，Western blot检测B细胞淋巴瘤-2相关X蛋白（B-cell lymphoma-2-associated X protein, Bax）、天冬氨酸蛋白水解酶3（cysteinyl aspartate specific proteinase-3, Caspase-3）、B细胞淋巴瘤-xl（B-cell lymphoma-xl, Bcl-xl）、环氧合酶-2（cyclooxygenase-2, COX-2）、基质金属蛋白酶-3（matrix metalloproteinase-3, MMP-3）和血小板结合蛋白基序的解聚蛋白样金属蛋白酶-5（recombinant a disintegrin and metalloproteinase with thrombospondin-5, ADAMTS-5）蛋白表达情况。结果 与正常组比较，模型组NPC活力显著下降（P<0.01），凋亡率增加（P<0.01），Bax、Caspase-3、COX-2、MMP-3和ADAMTS-5蛋白表达升高（P<0.05），Bcl-xl蛋白表达降低（P<0.05）；与模型组比较，不同浓度的ABS处理后，NPC活力显著上升（P<0.05），凋亡率减少（P<0.05），Bax、Caspase-3、COX-2、MMP-3和ADAMTS-5蛋白表达降低（P<0.05），Bcl-xl蛋白表达升高（P<0.05）。结论 ABS可以通过抑制由IL-1β诱导引起的NPC凋亡、炎症反应、ECM降解，促进细胞增殖和活性，对NPC产生保护作用，其机制可能与抑制Bax、Caspase-3、ADAMTS-5、MMP-3及COX-2表达，促进Bcl-xl表达相关。 |
| 关键词: 炎性损伤 牛膝总皂苷 髓核细胞 细胞凋亡 椎间盘退行性变性 |
| DOI：10.3969/j.issn.1674-070X.2023.09.007 |
| Received:April 10, 2023 |
| 基金项目:湖南省自然科学基金项目（2022JJ80086，2022JJ70029）；湖南省卫生健康委员会计划研究项目（D202302078705）；湖南省中医药管理局科研计划项目（B2023150，2021161，D2022094）；长沙市自然科学基金项目（kq2202460）。 |
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| Effects of achyranthes bidentata saponins on IL-1β-induced apoptosis and inflammatory injury of nucleus pulposus cells |
| LI Shuofu,LIU Hongzhe,ZHANG Jialin,YANG Lei,CHEN Long,GUO Yantao,LIU Xiaorong |
| (The First Hospital of Hunan University of Chinese Medicine, Changsha, Hunan 410007, China;Hunan Provincial People's Hospital, Changsha, Hunan 410002, China;Central South University, Changsha, Hunan 410083, China;Hunan University of Chinese Medicine, Changsha, Hunan 410208, China) |
| Abstract: |
| Objective To investigate the protective effects of achyranthes bidentata saponin (ABS) on interleukin-1β (IL-1β) induced injury in human nucleus pulposus cell (NPC) by observing the apoptosis, inflammation, and extracellular matrix (ECM) metabolism of NPC. Methods The degeneration model of NPC was established by IL-1β induction, the cells were randomly divided into normal group, model group (10 ng/mL IL-1β), low- (10 ng/mL IL-1β+3 μg/mL ABS), medium- (10 ng/mL IL-1β+10 μg/mL ABS), and high-dose (10 ng/ mL IL-1β+30 μg/mL ABS) ABS groups, and the intervention was performed for 24 h. Cell viability was determined by CCK-8 assay, and apoptosis was tested by Tunel assay and flow cytometry; Western blot was used to determine the protein expressions of B-cell lymphoma-2-associated X protein (Bax), cysteinyl aspartate specific proteinase-3 (Caspase-3), B-cell lymphoma-xl (Bcl-xl), cyclooxygenase-2 (COX-2), matrix metalloproteinase-3 (MMP-3), and recombinant a disintegrin and metalloproteinase with thrombospondin-5 (ADAMTS-5). Results Compared with the normal group, the model group showed significantly lower NPC viability (P<0.01), higher apoptosis rate (P<0.01), elevated protein expressions of Bax, Caspase-3, COX-2, MMP-3, and ADAMTS-5 (P<0.05), and reduced Bcl-xl protein expression (P<0.05); compared with the model group, after ABS treatment of different concentrations, NPC viability increased significantly (P<0.05), apoptosis rate was reduced (P<0.05), the protein expressions of Bax, Caspase-3, COX-2, MMP-3, and ADAMTS-5 were lower (P<0.05), and Bcl-xl protein expression was higher (P<0.05). Conclusion ABS has protective effects on NPCs by inhibiting apoptosis, inflammatory response, and ECM degradation of NPC induced by IL-1β and promoting cell proliferation and activity. Its mechanism may be related to the inhibition of Bax, Caspase-3, ADAMTS-5, MMP-3, and COX-2 expressions, and the promotion of Bcl-xl expression. |
| Key words: inflammatory injury achyranthes bidentata saponins nucleus pulposus cells apoptosis intervertebral disc degeneration |
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