KBN触变凝胶剂对非感染性炎症模型动物的抗炎作用及机制研究

Anti-inflammatory Effects of KBN Thixotropic Gel on Animal Models with Non-infective Inflammation and Its Mechanism

【目的】观察KBN触变凝胶剂(由黄连、龙胆等10味中药组成)的抗炎药效,并从核转录因子-κB(NF-κB)信号通路初步探讨其作用机制。【方法】将60只SD大鼠随机分为空白组,模型组,地塞米松组,KBN低、中、高剂量(生药质量浓度分别为0.1、0.2、0.4 g·mL-1)组,每组10只。采用右后足垫皮下注射角叉菜胶法建立大鼠足跖肿胀模型。测量各组大鼠不同时间点的足容积,计算足肿胀度,采用酶联免疫吸附分析(ELISA)法检测大鼠血清中肿瘤坏死因子α(TNF-α)、白细胞介素1β(IL-1β)的含量,蛋白质免疫印迹(Western blot)法检测大鼠足部软组织中NF-κB p65、核转录因子抑制蛋白(IκBα)、环氧合酶2(COX-2)的表达。另将60只KM小鼠随机分为空白组,模型组,地塞米松组,KBN低、中、高剂量组(生药质量浓度分别为0.1、0.2、0.4 g·mL-1),每组10只。采用涂布二甲苯法诱导小鼠耳廓肿胀模型。小鼠打耳片称质量,计算耳肿胀度,采用免疫组化(IHC)法检测小鼠耳廓组织中NF-κB p65、IκBα的表达水平。【结果】与模型组比较,KBN低、中、高剂量组大鼠足肿胀度在致炎后0.5~6 h降低,血清TNF-α、IL-1β含量显著降低,足部软组织NF-κB p65、COX-2表达降低、IκBα表达增加,均呈剂量依赖性。与模型组比较,KBN低、中、高剂量组小鼠耳肿胀度显著降低,耳廓组织中NF-κB p65表达降低,IκBα表达增加,均呈剂量依赖性。【结论】KBN触变凝胶剂具有良好的抗炎作用,其机制可能与抑制NF-κB信号通路活化进而降低炎症因子的释放有关。

Objective To observe the anti-inflammation effects of KBN Thixotropic Gel(composed of 10 Chinese medicinal materials involving Rhizoma Coptidis,Radix Gentianae,etc.),and to explore its mechanism based on nuclear factor(NF)-κB signaling pathway.Methods Sixty SD rats were randomly divided into 6 groups,namely blank group,model group,dexamethasone group,low-,middle-and high-dose KBN groups(at the mass concentrations of 0.1,0.2,0.4 g·mL-1 of crude drug,respectively),10 rats in each group.Paw edema rat model was established by subcutaneous injection ofλ-carrageen to right hind paw pad.The rat paw volume was measured at different time to calculate paw edema degree,the serum contents of tumor necrosis factor(TNF)-αand interleukin(IL)-1βwere detected by enzyme linked immunosorbent assay(ELISA),and the expression levels of NF-κB p65,inhibitor kappa B-alpha(IκB-α)and cyclooxygenase(COX)-2 in soft tissue of paw pad were detected by Western blotting method.Moreover,sixty KM mice were randomly divided into 6 groups,namely blank group,model group,dexamethasone group,high-,middle-and low-dose KBN groups(at the mass concentrations of 0.1,0.2,0.4 g·mL-1 of crude drug,respectively),10 mice in each group.Ear edema mouse model was established by external use of dimethyl benzene on ear.The ear slices were weighed to calculated ear edema degree,and the expression levels of NF-κB p65 and IκBαin ear tissue by detected by immunohistochemical method.Results Compared with the model group,the paw edema degrees of rats during 0.5-6 h after inflammation induction in low-,middle-and high-dose KBN groups were decreased,the serum contents of TNF-αand IL-1βwere reduced,the expression levels of NF-κB p65 and COX-2 were lowered,and IκB-αwas increased in soft tissue of paw pad,in dose-dependent manner.Compared with the model group,the ear edema degrees of mice in low-,middle-and high-dose KBN groups were decreased,and the expression level of NF-κB p65 was lowered and IκBαwas increased in auricular tissue,in dose-dependent manner.Conclusion KBN thixotropic gel has a marked anti-inflammatory effect,which possible mechanism is related with inhibiting the activation of NF-κB signaling pathway resulting in down-regulation of inflammatory factors.