女贞子多糖对IL-1β诱导的关节软骨细胞损伤和炎症反应的作用

Effects of Ligustrum lucidum polysaccharide on IL-1β-induced articular chondrocyte injury and inflammatory response

目的探讨女贞子多糖对白细胞介素1β(IL-1β)诱导的关节软骨细胞损伤和炎症反应的影响及分子机制。方法人关节软骨细胞分为对照组(不作任何处理),模型组(50 ng/mL IL-1β),女贞子多糖低、中、高剂量组(50、200、800μg/mL女贞子多糖),si-circ_0039411组,si-NC组,miR-145组,miR-NC组,女贞子多糖(800μg/mL)+pcDNA-circ_0039411组,女贞子多糖(800μg/mL)+anti-miR-145组。MTT法检测细胞活性,流式细胞术检测细胞凋亡,Western blot法检测蛋白表达,ELISA法检测IL-8、IL-6、IL-10水平,RT-qPCR法检测circ_0039411和miR-145的表达,双荧光素酶报告实验检测circ_0039411和miR-145的靶向关系。结果不同剂量女贞子多糖处理后的软骨细胞活性升高,细胞凋亡率降低,Bax蛋白表达降低,Bcl-2蛋白表达升高,IL-8、IL-6水平升高,IL-10水平降低,circ_0039411表达升高,miR-145表达降低,并呈剂量依赖性(P<0.05)。干扰circ_0039411或过表达miR-145,软骨细胞的活性升高,细胞凋亡率降低,Bax蛋白表达降低,Bcl-2蛋白表达升高,IL-8、IL-6水平降低,IL-10水平升高(P<0.05)。circ_0039411靶向调控miR-145;过表达circ_0039411或抑制miR-145可逆转女贞子多糖对IL-1β诱导的软骨细胞损伤。结论女贞子多糖可能通过调节circ_0039411/miR-145缓解IL-1β诱导的关节软骨细胞损伤和炎症反应。

AIM To explore the effects and molecular mechanism of Ligustrum lucidum polysaccharide on the injury and inflammatory response of articular chondrocytes induced by interleukin 1β(IL-1β).METHODS Human articular chondrocytes were divided into control group for no treatment,model group(50 ng/mL IL-1β),groups for intervention of low dose,medium dose and high dose L.lucidum polysaccharide(50,200,800μg/mL)respectively,si-circ_0039411 group,si-NC group,miR-145 group,miR-NC group,L.lucidum polysaccharide+pcDNA-circ_0039411 group,and L.lucidum polysaccharide+anti-miR-145 group.The articular chondrocytes were subsequently subjected to the detection of cell activity by MTT assay;cell apoptosis by flow cytometry;protein expressions by Western blot;IL-8,IL-6 and IL-10 levels by ELISA;and expressions of circ_0039411 and miR-145 by RT-qPCR.And the targeting relationship between circ_0039411 and miR-145 was determined by dual luciferase reporting assay.RESULTS L.lucidum polysaccharides treatments dose-dependently(P<0.05)increased chondrocyte activity and apoptosis rate,expressions of Bcl-2 protein and circ_0039411,and IL-8 and IL-6 levels;decreased Bax protein expression and miR-145 expression,and IL-10 levels.Interference with circ_0039411 or overexpression of miR-145 increased chondrocyte activity,decreased apoptosis rate,Bax protein expression,and IL-8 and IL-6 levels;increased Bcl-2 protein expression and IL-10 level(P<0.05).With circ_0039411 being the miR-145 target regulator,either circ_0039411 overexpression or miR-145 inhibition offset the effect of L.lucidum polysaccharide on IL-1β-induced chondrocyte injury.CONCLUSION L.lucidum polysaccharide may alleviate IL-1β-induced articular chondrocytes injury and inflammatory response via regulation on circ_0039411/miR-145.