高压氧联合儿茶素通过调控TRAF6/NF-κB信号通路抑制人骨髓基质HS-5细胞的炎症反应  

Hyperbaric oxygen combined with catechin inhibits the inflammatory response of human bone marrow stromal HS-5 cells by regulating TRAF6/NF-κB signaling pathway

作　　者：白左明[1] 何伟福 康明明 郭鹏 王东[1] Bai Zuoming;He Weifu;Kang Mingming;Guo Peng;Wang Dong(Department of Orthopedics,Second Hospital of Shanxi Medical University,Taiyuan 030053,China)

机构地区：[1]山西医科大学第二医院骨科,太原030053

出　　处：《中华航海医学与高气压医学杂志》2022年第5期652-655,660,共5页Chinese Journal of Nautical Medicine and Hyperbaric Medicine

摘　　要：目的探讨高压氧(HBO)联合儿茶素(Can)对脂多糖(LPS)诱导的人骨髓基质HS-5细胞炎症反应的影响及其作用机制。方法将一部分状态良好的正常HS-5细胞分为对照组(DMEM培养基),另一部分HS-5细胞随后通过LPS构建骨髓炎症细胞模型后再分为4组:模型组(培养基中加入1.0μg/ml LPS)、HBO组(培养基中加入1μg/ml LPS+0.2 MPa HBO处理)、Can组(培养基中加入1.0μg/ml LPS+150μmol/L Can)及联合组(培养基中加入1μg/ml LPS+150μmol/L Can+0.2 MPa HBO处理)。采用CCK-8法检测各组HS-5细胞的增殖情况,通过ELISA法测定HS-5炎症细胞液中IL-1β、IL-6、TNF-α和IL-10的表达水平,流式细胞术检测各组HS-5炎症细胞的凋亡情况,Western blotting实验检测HS-5炎症细胞TLR6/NF-κB信号通路蛋白的表达水平。结果CCK-8法检测结果显示,浓度150μmol/L Can对1.0μg/ml LPS诱导的HS-5细胞增殖产生抑制作用,并以此为参照建立骨髓HS-5炎症细胞模型。流式细胞术检测结果显示,联合组HS-5炎症细胞存活率[(13.99±1.25)%]明显低于模型组[(46.69±4.27)%]、HBO组[(39.28±3.74)%]和Can组[(23.14±2.20)%],差异均有统计学意义(P<0.05或P<0.01)。与模型组、HBO组和Can组比较,联合组HS-5炎症细胞内TLR6和p-NF-κB蛋白表达水平明显降低(P<0.01)。结论Can联合HBO对HS-5炎症细胞具有很强的抗炎作用,其作用机制可能是通过调节TLR6/NF-κB信号通路来抑制LPS诱导产生的骨髓炎症反应,这将为骨伤患者的治疗提供新的思路。Objective To discuss the effect of hyperbaric oxygen(HBO)combined with catechin(Can)on lipopolysaccharide(LPS)-induced inflammatory response of human bone marrow stromal HS-5 cells and its mechanism of action.Methods A portion of normal HS-5 cells in good condition were assigned as control group(DMEM medium),and another portion of HS-5 cells were used to establish the bone marrow inflammatory cell model induced by LPS which were then divided into four groups for different interventions:the model group(1.0μg/ml LPS was added into the medium),the HBO group(1.0μg/ml LPS was added into the medium and then treated with 0.2 MPa HBO),the Can group(1.0μg/ml LPS and 150μmol/L Can were added into the medium)and the combined group(1.0μg/ml LPS and 150μmol/L Can were added into the medium and then treated with 0.2 MPa HBO).The proliferation of HS-5 cells in each group was detected by the CCK-8 method,and the expression levels of IL-1β,IL-6,TNF-α,and IL-10 in HS-5 inflammatory cell fluid were measured by ELISA.The apoptosis of HS-5 inflammatory cells in each group was detected by flow cytometry,and the expression levels of TLR6/NF-κB signaling pathway proteins in HS-5 inflammatory cells were detected by western blotting.Results The results of the CCK-8 method showed that Can at the concentration of 150μmol/L inhibited the proliferation of HS-5 cells induced by LPS at 1.0μg/ml,which was used as a reference to establish an inflammatory cell model of bone marrow HS-5.The results of flow cytometry showed that the survival rate of HS-5 inflammatory cells in the combined group[(13.99±1.25)%]was significantly lower than that in the model group[(46.69±4.27)%],the HBO group[(39.28±3.74)%],and the Can group[(23.14±2.20)%],with statistically significant differences(P<0.05 or P<0.01).Compared with the model group,the HBO group,and the Can group,the expression levels of proteins(TLR6 and p-NF-κB)in HS-5 inflammatory cells in the combined group were significantly decreased(P<0.01).Conclusion Can combine with HBO has a stron

关 键 词：高压氧 儿茶素 TRAF6/NF-kB信号通路 人骨髓基质 HS-5细胞 炎症反应 

分 类 号：R68[医药卫生—骨科学]