活血化瘀利水方对急性踝关节损伤炎症大鼠P65、TAK1的影响

Effect of Huoxue Huayu Lishui Decoction on P65 and TAK1 of acute soft tissue injury in rats

目的:探讨活血化瘀利水方对急性踝关节损伤的作用机制。方法:将48只SPF级大鼠随机分为空白组、模型组、舒筋活络胶囊组及活血化瘀利水方低、中、高剂量组,其中空白组不予以造模处理,其余各组予以重锤坠落锤击造模法处理,每组8只。舒筋活络胶囊组予以0.2 g/mL混合液,低、中、高活血化瘀利水方组分别予以1.03、2.06、4.12 g/mL浓度水煎液,按1.5 mL/100 g灌胃;药物空白对照组、模型组蒸馏水3 mL灌胃,1 d 1次,连续1周。蛋白免疫印迹法检测大鼠踝关节周围软组织NF-κB P65、转化生长因子激酶1(TAK1)蛋白表达,运用PCR技术检测NF-κB P65、TAK1 mRNA表达。结果:造模后1 d,模型组、阳性药物组及低、中、高剂量活血化瘀利水方的P65、TAK1 mRNA及蛋白表达与空白对照组的差异有统计学意义(P<0.05)。服药7 d后,与模型组相比,舒筋活络胶囊组及低、中、高剂量活血化瘀利水方组服药后P65、TAK1 mRNA及蛋白的表达均降低(P<0.05)。用药7 d后,与舒筋活血胶囊组相比,低剂量活血化瘀利水方组服药后P65蛋白无差异,中、高剂量组与舒筋活血胶囊组P65蛋白的表达均降低(P<0.05),与舒筋活血胶囊组相比,中、高剂量组与舒筋活血胶囊组TAK1蛋白以及P65、TAK1 mRNA的表达均降低(P<0.05)。结论:活血化瘀利水方可能通过调控NF-κB大鼠踝关节损伤NF-κB P65、TAK1的表达,以改善踝关节损伤。

Objective:To explore the mechanism of the action of Huoxue Huayu Lishui decoction on acute ankle joint injury.Methods:Forty-eight SPF rats were randomly divided into blank group,model group,Shujin Huoluo capsule group,low,medium and high dose groups with Huoxue Huayu Lishui decoction.The blank group didn't receive any treatment,and the other groups were treated by hammering.Each group had 8 rats.Rats in the Shujin Huoluo capsule group was given mixed liquid of Shujin Huoluo capsule at 0.2 g/mL by gavage,and the low,medium and high dose groups with Huoxue Huayu Lishui decoction were given the decoction at 1.03 g/mL,2.06 g/mL,4.12 g/mL,respectively by gavage.The blank group and model group were given distilled water 3 mL by gavage.The gavage was performed once a day for 1 week.The expression of NF-κB P65 and transforming growth factor kinase 1(TAK1)protein in the soft tissue around the ankle joint of rats was detected by Western blotting,and the mRNA expression of NF-κB P65 and TAK1 was detected by PCR technology.Results:One day after modeling,the expressions of mRNA and protein of P65 and TAK1 in the model group,the positive drug group,and low,medium and high dose groups of Huoxue Huayu Lishui decoction were significantly different from those of the blank control group(P<0.05).Seven days after treatment,compared with the model group,the expressions of P65 and TAK1 mRNA and protein in the Shujin Huoluo capsule group and the low,medium and high dose groups of Huoxue Huayu Lishui decoction were all decreased(P<0.05).Compared with the Shujin Huoxue Capsule group,there was no difference in P65 protein in the low-dose Huoxue Huayu Lishui decoction group.The expression of P65 protein in the middle and high dose groups and the Shujin Huoxue Capsule group were all decreased(P<0.05).Compared with the Shujin Huoxue Capsule group,the expressions of TAK1 protein,mRNA of P65 and TAK1 in the low,medium,and high groups and the Shujin Huoxue Capsule group were all decreased(P<0.05).Conclusion:Huoxue Huayu Lishui decoction may regulate the expression of NF-κB P65 and TAK1 in rats with ankle joint injury,so as to improve the local swelling and pain in the ankle joint.