金骨莲胶囊对炎症模型大鼠的抗炎作用及机制研究

目的:研究金骨莲胶囊对炎症模型大鼠的抗炎作用及机制。方法:将48只大鼠随机分为空白对照组、模型组、金骨莲胶囊低、中、高剂量组(0.66、1.32、2.64 g/kg)和地塞米松组(阳性对照,0.945 mg/kg),每组8只。空白对照组和模型组大鼠灌胃等体积水,其余各组大鼠灌胃相应药物,每天给药2次,连续3天。末次给药后30 min,模型组和给药组大鼠腹腔注射脂多糖(10 mg/kg)以复制炎症模型。腹腔注射6 h后,于大鼠尾静脉取血,采用酶联免疫吸附试验检测大鼠血清中肿瘤坏死因子(TNF-α)、白细胞介素1β(IL-1β)、IL-6、前列腺素E(2 PGE2)的含量;测定大鼠肺组织湿质量/干质量(W/D)比值;采用苏木精-伊红染色法观察大鼠肺组织的病理学形态变化;采用逆转录-聚合酶链式反应法检测大鼠肺组织中TNF-α、IL-6、PGE2、IL-1βmRNA的表达水平;采用Western blot法检测大鼠肺组织中核因子κB(NF-κB)p65蛋白的磷酸化水平和NF-κB抑制蛋白α(IκBα)蛋白的表达水平。结果:与空白对照组比较,模型组大鼠血清中TNF-α、IL-1β、IL-6、PGE2的含量,肺组织W/D比值,肺组织中TNF-α、IL-1β、IL-6、PGE2 mRNA的表达水平和NF-κB p65蛋白磷酸化水平均显著升高,IκBα蛋白表达水平显著降低(P<0.05或P<0.01);大鼠肺组织中大量肺泡萎缩塌陷、壁增厚,可见肺实质化及大量炎症细胞浸润。与模型组比较,金骨莲胶囊各剂量组大鼠血清中TNF-α、IL-1β(低剂量组除外)、IL-6、PGE2的含量,以及肺组织中TNF-α(低剂量组除外)、IL-1β、IL-6(低、高剂量组除外)、PGE2 mRNA的表达水平均显著降低(P<0.05或P<0.01);金骨莲胶囊高剂量组大鼠肺组织W/D比值显著降低(P<0.05或P<0.01);金骨莲胶囊中剂量组大鼠肺组织中NF-κB p65蛋白磷酸化水平均显著降低(P<0.05或P<0.01),IκBα蛋白表达水平显著升高(P<0.05);大鼠肺泡结构较为清晰、壁轻度增厚,并见少量炎症细胞浸润。结论:金骨莲胶囊对炎症模型大鼠具有良好的抗炎作用,其作用机制可能与抑制NF-κB信号通路有关。

Study on Anti-inflammatory Effect and Mechanism of Jingulian Capsule on Inflammatory Model Rats

OBJECTIVE:To study the anti-inflammatory effect and mechanism of Jingulian capsule on inflammatory model rats.METHODS:Totally 48 rats were randomly divided into blank control group,model group,Jingulian capsule low-dose,medium-dose and high-dose groups(0.66,1.32,2.64 g/kg),dexamethasone group(positive control,0.945 mg/kg),with 8 rats in each group.Blank control group and model group were given constant volume of water intragastrically,and other groups were given relevant medicine intragastrocally,twice a day,for consecutive 3 days.Thirty minutes after last administration,model group and administration groups were given lipopolysaccharide(10 mg/kg)intraperitoneally to induce inflammatory model.Six hours after intraperitoneal injection,the contents of TNF-α,IL-1β,IL-6,PGE2 in serum were detected by ELISA.The wet to dry weight(W/D)ratio of lung tissue were determined.HE staining was used to observe the pathological changes of lung tissue.RT-PCR was used to detect the mRNA expression of TNF-α,IL-6,PGE2 and IL-1βin lung tissue.Western blot assay was used to detect the phosphorylation of NF-κB p65 protein and the expression of IκBαprotein in lung tissue.RESULTS:Compared with blank control group,the contents of TNF-α,IL-1β,IL-6 and PGE2 in serum,the W/D ratio of lung tissue,the expression of TNF-α,IL-1β,IL-6 and PGE2 mRNA and the phosphorylation level of NF-κB p65 protein in lung tissue of model group were significantly increased,and the expression of IκBαprotein was significantly decreased(P<0.05 or P<0.01);a large number of alveolar atrophy and collapse,alveolar wall thickening,lung consolidation,and a large number of inflammatory cell infiltration could be seen in lung tissue.Compared with model group,the contents of TNF-α,IL-1β(except for low-dose group),IL-6 and PGE2 in serum,as well as the expression of TNF-α(except for high-dose group),IL-1β,IL-6(except for low-dose,high-dose groups)and PGE2 mRNA in lung tissue were decreased significantly in Jingulian capsule groups(P<0.05 or P<0.01);the W/D ratio of lung tissue was decreased significantly in Jingulian high-dose group(P<0.05 or P<0.01);the expression of phosphorylation level of NF-κB p65 protein in lung tissue of Jingulian medium-dose group were decreased significantly(P<0.05 or P<0.01),while the expression of IκBαprotein was increased significantly(P<0.05);the alveolar structure was clear,the alveolar wall was slightly thickened,and a small amount of inflammatory cell infiltration was seen in lung tissue.CONCLUSIONS:Jingulian capsule has good anti-inflammatory effect on inflammatory model rats,the mechanism of which may be related to the inhibition of NF-κB signal pathway.