愈疡胶囊对细菌感染皮炎模型大鼠的抗炎作用及对TLR/NF-κB通路的影响
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篇名: 愈疡胶囊对细菌感染皮炎模型大鼠的抗炎作用及对TLR/NF-κB通路的影响
TITLE: Anti-inflammatory Effects of Yuyang Capsule on Bacterial Dermatitis Model Rats and Its Effect on TLR/NF-κB Signaling Pathway
摘要: 目的:研究愈疡胶囊对细菌感染皮炎模型大鼠的抗炎作用及对Toll样受体/核因子κB(TLR/NF-κB)通路的影响。方法:采用肉汤稀释法测定愈疡胶囊对金黄色葡萄球菌的最低抑菌浓度(MIC)和最低杀菌浓度(MBC)。将50只大鼠随机分为模型组、阳性对照组(阿莫西林克拉维酸钾,以阿莫西林计120mg/kg)和愈疡胶囊高、中、低剂量组(根据MIC设定),每组10只。以金黄色葡萄球菌感染大鼠烧伤皮肤构建细菌感染皮炎模型。造模成功24h后,各药物组大鼠灌胃相应药物,模型组大鼠灌胃等体积生理盐水,每日1次,连续7d。计算给药第1、3、5、7天各组大鼠皮肤愈合率并记录创面结痂、脱痂、愈合情况;使用酶联免疫吸附试验法检测皮肤组织中白细胞介素1β(IL-1β)、IL-6、IL-10、肿瘤坏死因子α(TNF-α)、羟脯氨酸(HYP)、胶原蛋白Ⅰ(ColⅠ)、ColⅢ的含量;使用苏木精-伊红染色法观察皮肤组织形态学变化,并使用透射电子显微镜观察超微结构;采用Westernblotting法检测皮肤组织中Toll样受体2(TLR2)、TLR4与磷酸化-NF-κB-p65(p-NF-κBp65)蛋白表达水平。结果:愈疡胶囊的MIC与MBC分别为25mg/mL和50mg/mL,愈疡胶囊高、中、低剂量组分别设为600、300、150mg/kg。与模型组比较,愈疡胶囊高、中剂量组大鼠给药3d后伤处皮肤出现结痂,给药5~7d后部分大鼠伤处出现脱痂;阳性对照组和愈疡胶囊高、中剂量组大鼠各时间点的皮肤愈合率均显著升高,IL-1β、IL-6、IL-10、TNF-α含量和病理评分以及TLR2、TLR4、p-NF-κBp65蛋白表达水平均显著降低(P<0.05或P<0.01),皮肤组织炎细胞浸润等病理改变好转;阳性对照组和愈疡胶囊高剂量组大鼠的HYP、ColⅠ、ColⅢ含量均显著升高(P<0.05或P<0.01)。而阳性对照组和愈疡胶囊高、中剂量组上述指标差异大多无统计学意义(P>0.05)。结论:愈疡胶囊可促进细菌感染皮炎模型大鼠皮肤愈合,具有一定的抗炎作用,其机制可能与抑制TLR/NF-κB信号通路、减少炎症反应有关。
ABSTRACT: OBJECTIVE:To study the anti-inflammatory effects of Yuyang capsule on bacterial dermatitis model rats and its effect on TLR/NF-κ B pathway. METHODS:Minimum inhibition concentration (MIC)and minimal bactericidal concentration (MBC)of Yuyang capsule against Staphylococcus aureus were determined by microdilution test. Totally 50 SD rats were randomly divided into model group ,positive control group (amoxicillin and clavulanate potassium ,120 mg/kg as amoxicillin ),Yuyang capsules high-dose ,medium-dose and low-dose groups (according to MIC ),with 10 rats in each group. The model of bacterial dermatitis was established by using the burned skin of rats infected with S. aureus . 24 h after modeling ,administration groups were intragastrically given the corresponding drug ,and model group was intragastrically given the same amount of normal saline ,once a day,for consecutive 7 days. The skin healing rate was calculated on the 1st,3rd,5th and 7th day of administration ,and the scab formation,decrustation and healing were recorded. The contents of IL- 1β,IL-6,IL-10,TNF-α,hydroxyproline(HYP),collagen Ⅰ(Col Ⅰ)and Col Ⅲ in skin tissue were detected by ELISA. Morphology changes of skin tissues were observed by HE staining. The ultrastructure was observed by transmission electron microscope. Protein expression of TLR 2,TLR4 and p-NF-κB p65 were detected by Western blotting assay. RESULTS :MIC and MBC of Yuyang capsule were 25 mg/mL and 50 mg/mL,respectively. Dose of Yuyang capsules high-dose ,medium-dose and low-dose groups were set at 600,300,150 mg/kg. Compared with model group,scab appeared on the injured skin 3 days after administration in Yuyang capsule high-dose and medium-dose groups ,and decrustation appeared on the injured skin of part mice 5-7 days after administration ;the skin healing rate of the positive control group,Yuyang capsule high-dose and medium-dose groups were all significantly increased at each time point. The contents of IL- 1 β,IL-6,IL-10 and TNF-α,pathological score ,protein expression of TLR 2,TLR4 and p-NF-κB p65 were decreased significantly (P<0.05 or P<0.01);the pathological changes such as inflammatory cell infiltration in skin tissue were improved. The contents of HYP,Col Ⅰ and Col Ⅲ were increased significantly in positive control group and Yuyang capsule high-dose group (P<0.05 or P<0.01). There was no statistical significance in most above indexes in positive control group ,Yuyang capsule high-dose and medium-dose groups (P>0.05). CONCLUSIONS :Yuyang capsule can promote skin healing of bacterial dermatitis model rats and shows certain anti-inflammatory effects ;the mechanism may be related to inhibiting TLR/NF-κB signaling pathway and reducing inflammatory response.
期刊: 2021年第32卷第04期
作者: 罗伦才,童妍,张兴国,陈礴,钟振东
AUTHORS: LUO Luncai ,TONG Yan,ZHANG Xingguo ,CHEN Bo,ZHONG Zhendong
关键字: 愈疡胶囊;细菌感染;感染性皮炎;Toll样受体/核因子κB信号通路;抑菌抗炎;大鼠
KEYWORDS: Yuyang capsules ;Bacterial infection ;Infectious dermatitis ;TLR/NF-κ B signaling pathway;Antibacterial and
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