低强度脉冲超声波对髁突软骨细胞炎症状态的作用研究

STUDY ON THE EFFECT OF LOW INTENSITY PULSED ULTRASOUND ON THE NEW STATE OF CONDYLAR CHONDROCYTES

目的 观察低强度脉冲超声波对白介素-1β制造的大鼠髁突软骨细胞炎症模型中MAPK信号通路相关因子磷酸化的表达变化。方法 二次酶消化法分离髁突软骨细胞并完成鉴定。将分离出的大鼠髁突软骨细胞随机分为3组,加入3种不同浓度的IL-1β处理并进行Ⅱ型胶原与MMP-13免疫细胞化学染色,筛选出最佳致炎浓度。再次将所提取的大鼠髁突软骨细胞随机分为3组:NC组、OA组、LIPUS组,处理后对Ⅱ型胶原、p38、p-p38、ERK1/2、pERK1/2进行免疫细胞化学染色,观察蛋白表达情况。结果(1)成功提取大鼠髁突软骨细胞。(2)IL-1β:10 ng/mL组Ⅱ型胶原表达量降低最明显,差异有统计学意义(P<0.05),MMP-13表达量升高最明显,差异有统计学意义(P<0.05)。(3)OA组与LIPUS组Ⅱ型胶原表达量均降低,差异有统计学意义(P<0.05),OA组降低更明显,差异有统计学意义(P<0.05)。3组的p38、ERK1/2表达量无明显差异(P>0.05)。OA组与LIPUS组p-p38、p-ERK1/2表达量均升高,差异有统计学意义(P<0.05),OA组升高更明显,差异有统计学意义(P<0.05)。结论(1)二次酶消化法成功提取大鼠髁突软骨细胞。(2)10 ng/mL为IL-1β最佳致炎浓度。(3)LIPUS处理可减少炎性髁突软骨细胞中Ⅱ型胶原的降解,抑制了MAPK通路相关因子p38、ERK1/2的磷酸化。

Objective To observe the effect of low-intensity pulsed ultrasound on the expression of phosphorylation of MAPK signaling pathway-related factors in the inflammatory model of rat condylar chondrocytes induced by interleukin-1β.Methods Condylar chondrocytes were isolated by secondary enzymatic digestion and identified.The isolated condylar chondro-cytes were randomly divided into 3 groups,and treated with three different concentrations of IL-1β,and immunocytochemistry staining of type II collagen and MMP-13 was performed to screen out the optimal inflammatory concentration.The extracted rat condylar chondrocytes were randomly divided into 3 groups:NC group,OA group and LIPUS group.After treatment,type II col-lagen,p38,p-p38,ERK1/2 and p-ERK1/2 were stained by immunocytochemistry to observe the protein expression.Results(1)Condylar chondrocytes were successfully isolated.(2)IL-1β:The expression of type II collagen in 10 ng/mL group decreased most significantly(P<0.05),and the expression of MMP-13 increased most significantly(P<0.05).(3)The expression of type II collagen in OA group and LIPUS group decreased(P<0.05),and the decrease in OA group was more obvious(P<0.05).There was no significant difference in the expression of p38 and ERK1/2 among the three groups(P>0.05).The expression of p-p38 and p-ERK1/2 in the OA group and the LIPUS group increased(P<0.05),and the increase in the OA group was more significant(P<0.05).Conclusions(1)Rat condylar chondrocytes were successfully extracted by secondary enzyme digestion.(2)10 ng/mL was the optimal concentration of IL-1β.(3)LIPUS treatment can reduce the degradation of type II collagen in in-flammatory condylar chondrocytes and inhibit the phosphorylation of MAPK pathway-related factors p38 and ERK1/2.