宝乐果提取物8,9-糖基化环烯醚萜苷促进人软骨细胞增殖和抑制凋亡的作用

目的观察宝乐果(Borojo)提取物8,9-糖基化环烯醚萜苷(GIG)对体外培养的人骨关节炎软骨细胞促进增殖和抑制细胞凋亡的作用。方法体外培养人骨关节炎软骨细胞,进行软骨细胞鉴定。用不同浓度(25、50、100、200、400μg/ml)的8,9-糖基化环烯醚萜苷培养人骨关节炎软骨细胞,用硫酸酚嗪甲酯(MTS)法、5-乙炔基-2脱氧尿嘧啶核苷(EdU)法、平板克隆形成实验检测细胞的增殖、流式细胞术观察软骨细胞凋亡,用丙二醛(MDA)法、超氧化物歧化酶(SOD)检测、谷胱甘肽过氧化物酶(GSH-Px)检测观察软骨细胞氧化酶的活性,用Western bolt法检测凋亡相关蛋白表达。结果与对照组相比,随GIG浓度增加,细胞活力升高,差异有统计学意义(P<0.01);细胞核分裂增加,差异有统计学意义(P<0.05);细胞克隆数量增多,差异有统计学意义(P<0.05);细胞周期的S期延长,细胞增殖活跃细胞凋亡率减少;MDA含量下降,差异有统计学意义(P<0.05)。SOD含量升高,差异有统计学意义(P<0.05);GSH-Px含量升高,差异有统计学意义(P<0.05)。结论 GIG可能对软骨细胞具有增殖的作用,而且在观察浓度内,细胞毒性小,并可抑制细胞凋亡,提示其有潜在能力可能成为治疗骨关节炎的一种策略。

Borojo extract 8,9-glycosylated iridoid glycosides promote the proliferation of human chondrocytes and inhibit apoptosis

Objective To observe the effects of Borojo extract 8,9-glycosylated iridoid glycosides(GIG)on humanosteoarthritis chondrocytes cultured in vitro to promote proliferation and inhibit cell apoptosis.MethodsHuman osteoarthri-tis chondrocytes were cultured in vitro for chondrocyte identification. Culture human osteoarthritis chondrocytes with differentconcentrations(25,50,100,200,400 μg/ml)of 8,9-glycosylated iridoid glycosides,using MTS method,Ed U detec-tion,plate clone formation experiment,flow cytometry Cytometry was used to observe the proliferation and apoptosis of chon-drocytes,the activity of chondrocyte oxidase was detected by MDA,SOD,and GSH-Px,and the expression of apoptosis-re-lated proteins was detected by Western blot.ResultsCompared with the control group detected by MTS,the cell viabilityincreased,which was statistically significant(P<0.01). Ed U detection,compared with the control group,increased cell nu-clear division,which was statistically significant(P<0.05). In plate clone detection,compared with the control group,thenumber of cell clones increased,which was statistically significant(P<0.05). Flow cytometry detected the cell cycle. Com-pared with the control,the s phase of the cell cycle was prolonged and the cell proliferation was active. Flow cytometry wasused to detect cell apoptosis. Compared with the control group,the apoptosis rate was reduced. MDA detects the antioxidanteffect of cells. Compared with the control group,the MDA content decreased,which was statistically significant(P<0.05).Compared with the control group,the SOD detection increased the SOD content,which was statistically significant(P<0.05). Compared with the control group,the GSH-Px test showed that the GSH-Px content increased,which was statisticallysignificant(P<0.05).Conclusion8,9-glycosylated iridoid glycosides have a proliferation effect on chondrocytes,andwithin the observed concentration,there is no cytotoxicity. 8,9-glycosylated iridoid glycosides can inhibit cell apoptosis. 8,9-glycosylated iridoid glycosides have the potential to become methods for the treatment of osteoarthritis.