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Efficacy of Andrographis paniculata against extended spectrum β-lactamase (ESBL) producing E. coli

机译:穿心莲对产超广谱β-内酰胺酶(ESBL)的大肠杆菌的功效

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A. paniculata is widely known for its medicinal values and is traditionally used to treat a wide range of diseases such as cancer, diabetes, skin infections, influenza, diarrhoea, etc. The phytochemical constituents of this plant possess unique and interesting biological activities. The main focus of this study was to evaluate the antibacterial property of crude ethyl acetate (CEA) extract of A. paniculata against E. coli clinical isolates along with molecular docking of 10 different bioactive components from this plant with CTX-M-15. CEA extract was subjected to phytochemical and FTIR analysis. The E. coli isolates were tested for antibiotic susceptibility through disk-diffusion method to observe their resistance pattern towards different antibiotics. Antibacterial activity and biofilm assay were performed through broth microdilution using a 96-well microplate. CEA extract was further utilized to observe its effect on the expression of a gene encoding CTX-M-15. Finally, in-silico studies were performed where 10 different bioactive compounds from A. paniculata were molecularly docked with CTX-M-15. Phytochemical and FTIR analysis detected the presence of various secondary metabolites and functional groups in CEA extract respectively. Molecular docking provided the number of residues and bond lengths together with a positive docking score. Antibiotic susceptibility showed the multi-drug resistance of all the clinical strains of E. coli. The antibacterial and antibiofilm efficiency of CEA extract (25, 50 and 100?μg/ml) was tested and 100?μg/ml of the extract was more effective in all the strains of E. coli. All 3 ESBL producing strains of E. coli were subjected to gene expression analysis through PCR. Strains treated with 100?μg/ml of the extract showed a downregulation of the gene encoding CTX-M-15 compared to untreated controls. The utilization of CEA extract of A. paniculata proved an economical way of controlling the growth and biofilm formation of ESBL strains of E. coli. CEA extract was also able to downregulate the expression of a gene encoding CTX-M-15. Molecular docking of 10 different bioactive compounds from A. paniculata with CTX-M-15 provided the residues and bond lengths with a positive docking score.
机译:Paniculata因其药用价值而广为人知,并且传统上用于治疗多种疾病,例如癌症,糖尿病,皮肤感染,流感,腹泻等。该植物的植物化学成分具有独特而有趣的生物学活性。这项研究的主要重点是评估paniculata的粗乙酸乙酯(CEA)提取物对大肠杆菌临床分离株的抗菌性能,以及该植物中10种不同生物活性成分与CTX-M-15的分子对接。对CEA提取物进行植物化学和FTIR分析。通过圆盘扩散法测试了大肠杆菌分离物的抗生素敏感性,以观察其对不同抗生素的抗药性。通过使用96孔微孔板的肉汤微量稀释进行抗菌活性和生物膜测定。进一步利用CEA提取物观察其对编码CTX-M-15的基因表达的影响。最后,进行了计算机模拟研究,其中将10种不同的来自A. paniculata的生物活性化合物与CTX-M-15分子对接。植物化学和FTIR分析分别检测到CEA提取物中各种次生代谢物和官能团的存在。分子对接提供了残基的数目和键长以及对接得分为正。抗生素敏感性显示所有临床大肠杆菌菌株均具有多重耐药性。测试了CEA提取物(25、50和100?μg/ ml)的抗菌和抗生物膜效率,并且100?μg/ ml的提取物在所有大肠杆菌中均更有效。通过PCR对所有3株产生ESBL的大肠杆菌菌株进行基因表达分析。与未处理的对照相比,用100?μg/ ml的提取物处理的菌株显示出编码CTX-M-15的基因的下调。沙棘曲霉CEA提取物的利用被证明是控制大肠杆菌ESBL菌株生长和生物膜形成的经济方法。 CEA提取物还能够下调编码CTX-M-15的基因的表达。 10种不同生物活性化合物与A. paniculata的分子对接,其CTX-M-15的残基和键长为对接分数。

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