摘要
目的探究如意金黄散对耐甲氧西林金黄色葡萄球菌(MRSA)感染创面炎症及Toll样受体(TLR)-核因子κB(NF-κB)通路的影响。方法将41只清洁级SD雌性大鼠随机分为生理盐水组10只、75%酒精组10只、莫匹罗星组10只、金黄散酊组11只。各组均制备MRSA感染创面,造模成功后次日,生理盐水组、75%酒精组、莫匹罗星组、金黄散酊组分别给予生理盐水、75%酒精、莫匹罗星软膏、如意金黄散酊换药,每日1次,连续14 d。记录治疗第1天、第3天、第7天、第14天创面面积,测定创面菌落计数,采用ELISA法检测肉芽组织中NF-κB p65、白细胞介素-6(IL-6)、白细胞介素-10(IL-10)含量,采用RT-PCR法检测治疗第7天肉芽组织中1-TLR2、1-TLR4 mRNA表达情况。结果MH羊血琼脂培养平板菌落计数,金黄散酊组治疗第3天和第14天均明显低于生理盐水组(P均<0.05);MRSA显色培养平板菌落计数,莫匹罗星组治疗第3天和第7天均明显低于其他3组(P均<0.05),金黄散酊组治疗第7天明显低于75%酒精组(P<0.05),莫匹罗星组治疗第14天明显低于75%酒精组与金黄散酊组(P均<0.05)。治疗第3天,金黄散酊组NF-κB p65含量明显高于75%酒精组(P<0.05),IL-6含量明显低于莫匹罗星组(P<0.05);治疗第7天,金黄散酊组IL-10含量明显高于生理盐水组及75%酒精组(P均<0.05),莫匹罗星组IL-10含量明显高于生理盐水组(P<0.05);治疗第14天,金黄散酊组和莫匹罗星组IL-10含量均明显高于生理盐水组及75%酒精组(P均<0.05)。治疗第7天,莫匹罗星组和金黄散酊组1-TLR2、1-TLR4 mRNA相对表达量均明显低于生理盐水组(P均<0.05),且金黄散酊组1-TLR2、1-TLR4 mRNA相对表达量均明显低于75%酒精组(P均<0.05)。结论如意金黄散对于大鼠MASR感染创面的影响主要体现在非单一菌落中多样共生菌的体外抑制作用中,而在单一MRSA菌落生长的环境中,如意金黄散对其抑制作用并不明显。如意金黄散具有控制MRSA感染创面炎症的作用,其机制可能与抑制TLR-NF-κB通路的1-TLR2、1-TLR4受体的表达,从而调控炎症因子表达有关。
Objective It is to investigate the effects of Ruyi Jinhuang Powder(RJP)on inflammation of methicillin-resistant Staphylococcus aureus(MRSA)infected wounds and Toll-like receptor(TLR)-nuclear factor-κB(NF-κB)pathway.Methods Forty-one clean-grade SD female rats were randomly divided into normal saline group(n=10),75%alcohol group(n=10),mupirocin group(n=10),and RJP tincture group(n=11).MRSA-infected wounds were prepared in each group,and on the next day after successful modeling,the normal saline group,75%alcohol group,mupirocin group,and RJP tincture group were respectively treated with normal saline,75%alcohol,mupirocin ointment,and RJP tincture for dressing change,once daily,continuously treated for 14 days.The area of the wounds was recorded on the first,third,seventh,and fourteenth days of treatment,and the colony counts of wounds were determined,the contents of NF-κB p65,interleukin-6(IL-6),interleukin-10(IL-10)in the granulation tissue were detected by ELISA assay,and the expressions of 1-TLR2,1-TLR4 mRNA in the granulation tissue on the 7th day of treatment were detected by RT-PCR.Results The colony counts in MH sheep blood agar culture plate in the RJP tincture group on the 3rd and 14th day of treatment were significantly lower than those in the normal saline group(all P<0.05);the colony counts in MRSA chromogenic culture plate in the mupirocin group on the 3rd and 7th day of treatment were significantly lower than the other three groups(all P<0.05),the counts in the RJP tincture group on the 7th day of treatment was significantly lower than that in the 75%alcohol group(P<0.05),and the counts in the mupirocin group on the 14th day of treatment was significantly lower than the 75%alcohol group and RJP tincture group(both P<0.05).On the 3rd day of treatment,the content of NF-κBp65 in the RJP tincture group was significantly higher than that in the 75%alcohol group(P<0.05),and the content of IL-6 was significantly lower than that in the mupirocin group(P<0.05);on the 7th day of treatment,the content of IL-10 in the RJP tincture group was significantly higher than that in the normal saline group and 75%alcohol group(both P<0.05),and the content of IL-10 in the mupirocin group was significantly higher than that in the normal saline group(P<0.05);on the 14th day of treatment,the contents of IL-10 in both the RJP tincture group and mupirocin group were significantly higher than those in the normal saline group and 75%alcohol group(all P<0.05).On the 7th day of treatment,the relative expressions of 1-TLR2 and 1-TLR4 mRNA in both the mupirocin group and RJP tincture group were significantly lower than that in the normal saline group(P<0.05),and the relative expressions of 1-TLR2 and 1-TLR4 mRNA in the RJP tincture of Jinhuang San group were significantly lower than that in the 75%alcohol group(all P<0.05).Conclusion The effect of Ruyi Jinhuang Powder on MASR infected wounds in rats is mainly reflected in the inhibitory effect of diverse symbiotic bacteria in non-single colonies in vitro,while the inhibitory effect of Ruyi Jinhuang Powder on the growth of single MRSA colonies is not significant.Ruyi Jinhuang Powder could control the inflammation in MRSA-infected wounds,and its mechanism may be related to inhibiting the expression of 1-TLR2 and 1-TLR4 receptors of the TLR-NF-κB pathway,thus to regulate the expression of inflammatory factors.
作者
刘现周
戴睿
徐强
LIU Xianzhou;DAI Rui;XU Qiang(The Second Affiliated Hospital of Tianjin University of Traditional Chinese Medicine,Tianjin 300250,China;Tianjin University of Traditional Chinese Medicine,Tianjin 301617,China)
出处
《现代中西医结合杂志》
CAS
2023年第16期2191-2196,2208,共7页
Modern Journal of Integrated Traditional Chinese and Western Medicine
基金
国家自然科学基金青年科学基金项目(81904211)。
