莱菔硫烷降低尿酸钠诱导成骨细胞的炎症和氧化应激反应  

Sulforaphane reduces osteoblast damage and oxidative stress induced by MSU

作　　者：陈巧凤[1] 施建辉[1] 罗昌林 CHEN Qiaofeng;SHI Jianhui;LUO Changlin(Department of Orthopedics,the Affiliated Quanzhou First Hospital of Fujian Medical University,Quanzhou 362000;Fujian Medical University,Fuzhou 362002,China)

机构地区：[1]福建医科大学附属泉州第一医院骨科,福建泉州362000 [2]福建医科大学,福建福州362002

出　　处：《中国骨质疏松杂志》2023年第11期1598-1602,共5页Chinese Journal of Osteoporosis

基　　金：福建省自然科学基金项目(2021J11406)。

摘　　要：目的观察尿酸钠盐(monosodium urate,MSU)对成骨细胞的损伤,并探讨莱菔硫烷(sulforaphane,SFN)对MSU诱导的成骨细胞损伤的机制及治疗作用。方法用CCK-8试剂盒检测成骨细胞活力,用流式细胞仪检测成骨细胞凋亡,用ROS敏感荧光探针测定成骨细胞ROS水平。在成骨细胞培养基中分别加入MSU(10、50、100、300、500μg/mL)和SFN(1、5、10、20、50μmol/L)后0、12、24、48、72 h分别检测,确定药物的作用浓度和作用时间,用蛋白质印迹法检测成骨细胞Nrf2、HO-1、HIF-1、IL-6和caspase-3。结果MSU对成骨细胞的毒性作用随着加入作用浓度和作用时间的增加而增加。当MSU作用浓度为300μg/mL,作用时间超过48 h,细胞活力低于50%。当SFN在作用浓度为1μmol/L和10μmol/L时对成骨细胞的活力没有影响,但作用浓度为20μmol/L和50μmol/L时细胞活力明显降低。确定MSU作用浓度为300μg/mL,SFN作用浓度为10μmol/L,作用时间为48 h。MSU增加了成骨细胞内的ROS水平,SFN降低了MSU增加的成骨细胞内ROS水平和凋亡率。SFN降低了MSU诱导的成骨细胞的HIF-1、IL-6和caspase-3的表达水平,但增加了Nrf2、HO-1的表达水平。结论SFN可减少MSU对成骨细胞的损伤。Objective To observe the damage of MSU(monosodium-urate)on osteoblasts and the mechanism and therapeutic effect of SFN(sulforaphane)on osteoblasts induced by MSU.Methods CCK-8 kit was used to detect osteoblast viability.Flow cytometry was used to detect the apoptosis.ROS levels were confirmed using ROS sensitive fluorescent probe.Drug concentration and duration of action were determined after MSU(10,50,100,300,500μg/mL)and SFN(1,5,10,20,50μmol/L)were added respectively in osteoblasts culture medium for 0,12,24,48,72 h.Nrf2,HO-1,HIF-1,IL-6,and caspsae-3 in osteoblasts were detected using Western blotting.Results The toxic effect of MSU on osteoblasts activity increased along with the increase of concentration and time.Cell viability was less than 50%when the concentration was 300μg/mL and reaction time exceeded 48 hours.There was no effect on osteoblast viability when SFN was at 1μmol/L and 10μmol/L,but cell viability reduced when SFN was at 20μmol/L and 50μmol/L.300μg/mL and 10μmol/L were determined as action concentrations of MSU and SFN,respectively.48 h was determined as action time.MSU increased the level of ROS in osteoblasts,while SFN decreased the ROS level and apoptosis rate in osteoblasts increased by MSU.SFN reduced the expression levels of HIF-1,IL-6,and caspase-3 in MSU induced osteoblasts,but increased the expression levels of Nrf2 and HO-1.Conclusion SFN reduces osteoblast damage induced by MSU.

关 键 词：莱菔硫烷 尿酸钠盐 成骨细胞 痛风 氧化应激 

分 类 号：R965[医药卫生—药理学]