壳聚糖介导细胞因子反应调节因子A对软骨细胞白细胞介素-1β和肿瘤坏死因子α表达的抑制作用

Inhibitory effect of chitosan-mediated cytokine response modifier A on interkin-1β and tumor necrosis factor-α expression of chondrocytes

目的观察壳聚糖介导细胞因子反应调节因子A（CrmA）对软骨细胞白细胞介素（IL）-1β和肿瘤坏死因子（TNF）-α表达的作用。方法体外培养兔关节软骨细胞，分别加入磷酸盐缓冲液（PBS）、10mg／L壳聚糖和壳聚糖／pCDNA3．1^＋CnnA微粒处理6h后，加入10μg/L IL-1β共培养48h，采用实时定量聚合酶链反应（Real-time PCR）方法检测软骨细胞中IL-1β和TNF-αmRNA的表达。结果IL-1βmRNA在壳聚糖／pCDNA3．1^＋CrmA处理组（0．55±0．08）软骨细胞中的表达明显低于壳聚糖组（0．69±0．06，P〈0．01）和PBS组（0．99±0．04，P〈0．01），壳聚糖组软骨细胞IL-1β的表达与PBS组比较差异有统计学意义（P〈0．01）。TNF—αmRNA在壳聚糖／pCDNA3．1^＋CrmA处理组（0．57±0．07）软骨细胞中的表达与壳聚糖组（0．71±0．05）和PBS组（0．99±0．06）比较均显著下降（P〈0．01），壳聚糖处理组TNF-α的表达与PBS组比较差异有统计学意义（P〈0．01）。结论壳聚糖介导CrmA能够明显抑制IL-1β诱导的软骨细胞IL-1β和TNF-α的表达。

Objective To investigate the effects of chitosan/pCDNA3.1 ^＋ cytokine response modifier A （CrmA） nanoparticles on interkin-1β （IL-1β） and tumor necrosis factor-α （TNF-α） expression of chondroeytes. Methods Rabbit chondrocytes were isolated and cultured, and treated with PBS, 10 mg/L chitosan （CS） or chitosan/pCDNA3.1 ＋ CrmA nanoparticles, respectively for 6 h. Then 10 μg/L IL-1β was added into the culture medium. After 48 h, the mRNA expression of IL-1 β and TNF-α in chondrocytes was detected by using real-time polymerase chain reaction. Results In chitosan/pCDNA3.1 ^＋ CrmA-treated group （0. 55 ± 0. 08 ） , the mRNA expression of IL-1 β in chondrocytes was significantly suppressed as compared with corresponding samples of chitosan-treated group （ 0. 69 ±0. 06, P 〈 0. 01 ） and PBS-treated group （0. 99 ±0. 04,P 〈 0. 01 ）. There was significant difference in the IL-1β expression between chitosantreated group and PBS-treated group （ P 〈 0. 01 ）. The TNF-α mRNA expression of chondroeytes in chitosan/pCDNA3.1 ＋ CrrnA-treated group （0. 57 ± 0. 07 ） was lower than that in ehitosan-treated group （0. 71 ± 0. 05, P 〈 0. 01 ） and PBS-treated group （ 0. 99 ± 0. 06, P 〈 0. 01 ）. Significant difference of TNF-α expression between chitoson treated group and PBS treated group was observed （ P 〈 0. 01 ）. Conclusion CrmA mediated by ehitosan could significantly suppress the mRNA expression of IL-1 β and TNF-α of chondrocytes induced by interkin-1 β. chitosan/pCDNA3.1 CrmA nanoparticles maybe a potential candidate for therapy of osteoarthritis.