柚皮苷抑制胶原诱导小鼠关节炎症作用机制的实验研究

Experimental study of inhibitory effect and mechanism of Naringin on collagen-induced arthritis in mice

目的检测柚皮苷干预后胶原诱导(CIA)小鼠关节炎症相关指标的变化。方法选择雄性DBA1/J小鼠35只,随机分为正常组(5只)和实验组(30只)。实验组炎症评分达2分以上再分为骨碎补组(10只)、CIA模型组(10只)、柚皮苷组(10只)。各组均以灌胃方式给药,每天1次,连续20次。给药20 d后处死小鼠,ELISA方法测定小鼠血清中IL-6、TNF-α的水平,HE染色观察滑膜炎症表达情况,免疫组化法检测小鼠膝关节滑膜组织中HIF-1α、CXCR4蛋白的表达情况。结果给药第11天开始,柚皮苷组炎症评分低于CIA模型组,差异有统计学意义(P<0.05);给药第15天开始,柚皮苷组与骨碎补组炎症评分均低于CIA模型组,差异有统计学意义(P<0.05);给药第19天,骨碎补组与柚皮苷组炎症评分均有降低表现,且柚皮苷组炎症评分较骨碎补组低,差异有统计学意义(P<0.05)。柚皮苷、骨碎补组较CIA模型组的IL-6、TNF-α水平均下降,差异有统计学意义(P<0.01)。柚皮苷组的IL-6水平较骨碎补组低,TNF-α水平较骨碎补组高,但差异无统计学意义(P>0.05)。柚皮苷组HIF-1α、CXCR4的IOD值低于CIA模型组,差异有统计学意义(P<0.05);骨碎补组HIF-1α、CXCR4的IOD值与CIA模型组比较差异无统计学意义(P>0.05);骨碎补组与柚皮苷组2组间差异无统计学意义(P>0.05)。柚皮苷、骨碎补组染色阳性区域Area值均低于CIA模型组,差异有统计学意义(P<0.05);骨碎补组与柚皮苷组2组间比较差异无统计学意义(P>0.05)。结论柚皮苷对HIF-1α上游的炎症因子IL-6、TNF-α及下游的CXCR4都具有一定调控作用,显著的降低了CIA小鼠炎症反应。

Objective To detect the changes of relevant index in CIA mice by intervention of Naringin. Methods Thirty- five male DBA1/J mice were randomly divided into normal group （5 rats） and experimental group （30 rats）. When the inflammatory score was up to 2 points, experimental group was subdivided for Drynaria group（n =10）, the CIA model group（n = 10）, and Naringin group （n =10）. All groups were administered orally once a day, for 20 consecutive times. All the mice were sacrificed on 20th day after the drug administration. ELISA method was used for determination of the level of IL-6 and TNF-a in serum. HE staining was used to observe the synovitis expression, immune group of knee joint synovial tissue of mice was detected in HIF-lct and CXCR4 protein expression. Results From the llth day after administration of, inflammation score of Naringin group was lower than CIA model group, the difference was statistically significant （P 〈0.05）; From the 15th day after administration, inflammation score of Naringin group and Drynaria group were lower than the CIA model group, the difference was statistically significant （P 〈0.05）; On the 19th day after administration, inflammation score of Drynaria group and Naringin group inflammation were decreased, and Naringin group lower than the Drynaria group, the difference was statistically significant （P 〈0.05）. The levels of IL-6, TNF-a in Naringin and Drynaria group were lower than CIA model group, the difference was statistically significant （P 〈0.01）. IL-6 levels of Naringin group were lower than Drynaria group, TNF-a levels of Drynaria group higher than Naringin group, but the difference was not statistically significant （P 〉0.05）. IOD value of HIF-lct, CXCR4 in Naringin group were lower than CIA model group, the difference was statistically significant （P 〈0.05）, there was no significant difference between Drynaria group and CIA model group （P 〉0.05）,there was no significant difference between Drynaria group and Naringin group （P 〉0.05）. Area value of HIF-lot, CXCR4 in Naringin group and Drynaria group were lower than those in CIA model group, the difference was statistically significant （P 〈 0.05）, there was no significant difference between Drynaria group and Naringin group（P 〉0.05）. Conclusion Naringin plays a role of regulation in the upstream （IL-6 and TNF-a and downstream （CXCR4）of the HIF-1a, and it can reduce the inflammatory reaction of CIA mice significantly.