番石榴叶提取物对抵抗素诱导的人膝关节软骨细胞炎症的缓解作用及机制研究

Remission effect of guava leaf extract on resistin-induced chondrocytic inflammation of human knee chondrocytes and its mechanism

目的:探讨番石榴叶提取物(GLE)对抵抗素诱导的人膝关节软骨细胞炎症的缓解作用及机制。方法:对人膝关节软骨细胞进行培养,并将其分为对照组、抵抗素组及不同浓度(30、60、120μg/ml)GLE组。使用500 ng/ml抵抗素诱导分离的软骨细胞。通过实时荧光定量(RT-qPCR)检测细胞微小RNA-181a-5p(miR-181a-5p)表达水平。通过CCK-8实验检测各组细胞增殖情况。通过流式细胞术和Western blot检测各组细胞凋亡率及凋亡蛋白表达情况。通过酶联免疫吸附试验(ELISA)检测各组细胞白细胞介素6(IL-6)、肿瘤坏死因子(TNF)-α和TNF-β1表达情况。结果:与对照组比较,抵抗素组软骨细胞miR-181a-5p表达水平升高(P<0.05)。与抵抗素组比较,60、120μg/ml GLE组软骨细胞miR-181a-5p表达水平降低(均P<0.05)。与对照组比较,抵抗素组细胞OD值降低(P<0.05)。与抵抗素组比较,GLE组细胞OD值升高(P<0.05)。与对照组比较,抵抗素组细胞凋亡率、裂解形式的半胱氨酸蛋白酶蛋白-3(Cl-caspase-3)及B淋巴细胞瘤-2(Bcl-2)关联X蛋白(Bax)表达水平升高,Bcl-2蛋白表达水平降低(均P<0.05)。与抵抗素组比较,GLE组细胞凋亡率、Cl-caspase-3及Bax蛋白表达水平降低,Bcl-2蛋白表达水平升高(均P<0.05)。与对照组比较,抵抗素组IL-6、TNF-α及TNF-β1水平升高(均P<0.05)。与抵抗素组比较,GLE组IL-6、TNF-α及TNF-β1炎症因子水平降低(均P<0.05)。结论:番石榴叶提取物可缓解抵抗素诱导的人膝关节软骨细胞炎症,其机制可能与降低miR-181a-5p表达有关。

Objective:To investigate the effect of guava leaf extract(GLE)on resistin-induced chondrocytic inflammation of human knee chondrocytes and its mechanism.Methods:Human knee chondrocytes were cultured and divided into control group,resistin group,and GLE groups at various concentrations(30,60,120μg/ml).Isolated chondrocytes were induced by using 500 ng/ml resistin.The expression level of miR-181a-5p was measured by RT-qPCR.Cell proliferation was assessed by CCK-8.Cell apoptosis rate and apoptotic protein expression were determined by flow cytometry and Western blot.Expression of IL-6,TNF-αand TNF-β1 by ELISA.Results:The level of miR-181a-5p was increased in resistin group compared with control group(P<0.05).The level of miR-181a-5p expression was reduced in 60 and 120μg/ml GLE groups compared with resistin group(all P<0.05).The value of OD was reduced in resistin group compared with control group(P<0.05).The value of OD was higheRin GLE group than that in resistin group(P<0.05).Resistin group had increased apoptosis rate,Cl-caspase-3 and Bax protein levels and decreased Bcl-2 protein level compared with control group(all P<0.05).The apoptosis rate,Cl-caspase-3,and Bax protein levels were reduced and Bcl-2 protein level was increased in GLE group compared with resistin group(all P<0.05).IL-6,TNF-αand TNF-β1 levels were increased in resistin group compared with control group(all P<0.05).The levels of IL-6,TNF-αand TNF-β1 were loweRin GLE group than those in resistin group(all P<0.05).Conclusion:GLE can alleviates resistin-induced human knee chondrocyte inflammation,and its mechanism may be related to decreased miR-181a-5p expression.