Study On The Mechanism Of Chaiqintonglin Inhibiting Urinary System Infection Caused By Drug-Resistant Bacteria

Background: Urinary tract infection is the inflammatory response of the urothelial tract to the invasion of microorganisms such as bacteria.It usually refers to urinary tract and bladder infection.Escherichia coli is the most common pathogenic bacteria in urinary tract infection.In recent years,with the unscientific use of antibiotics with high frequency,increasing use of immunosuppressant drugs,and increasing clinical application of hormone drugs,the incidence of urinary system infection has increased significantly,and most of the clinically isolated strains of urinary infection are multidrug resistant strains,which increases the difficulty of clinical treatment of patients with urinary system infection.Therefore,how to inhibit the urinary tract infection caused by drug-resistant bacteria in clinic has become a research hotspot.Traditional Chinese medicine has a long history of use and rich experience in clinical use.Literature has shown that traditional Chinese medicine has antibacterial and immunological functions,providing an effective treatment for urinary system infections caused by drug-resistant bacteria in clinical practice.Chaiqintonglin(CQTL)are composed of Bupleurum,Radix Scutellariae,Polygonum and Dianmai,etc.According to the guidance of the basic theory system of traditional Chinese medicine,it has the effect of clearing away heat and detoxification,diuresis and drenching.It is mainly used in the clinic for urinary tract infection with good effect.This paper mainly studied the mechanism of Chaiqintonglin Tablets in inhibiting the urinary system infection caused by drug-resistant bacteria from the following parts:1.Effect of Chaiqin Tonglin Tablet on Suspend Bacteria Isolated from Clinical Drug Resistant Escherichia coliTrace drug broth dilution method determination of the experimental results show that the minimum bacteriostasis concentration experiment select 20 strains of clinical isolated urinary tract pathogenic e.coli(E5A2 E5A4 E5A6,E5A7,E5A11,E5A12,E5B2,E5B4,E4B5 / E5B5,E4B4,E4B6,E4C3,E4C8,E4C12,E6B2,E6B3,E6B4,E6B5 E5E5 E5E7)for clinical commonly used cephalosporins,aminoglycoside and quinolone basic performance for drug resistance,Cefoxitin and furantoin showed moderate drug sensitivity in most cases.Chaiqintonglin Tablets at 0.24g/ m L had inhibitory effect on 9 of 20 pathogenic Escherichia coli strains(E5A2,E5A11,E5B5,E4B6,E5A4,E5A7,E5B4,E5E5,E4C8).The results of the combined action of Chaiqintonglin and common antibiotics by chessboard method showed that the combined action of Chaiqintonglin and ceftazidime,amtronam,tigecycline,amikacin and gentamicin had no effect.In combination with ciprofloxacin can produce antagonistic effect.The results showed that after the continuous treatment of 1/4 MIC Chaiqintonglin tablets on 20 strains of Escherichia coli,3 of them showed recovery of susceptibility to ceftazidime.The MIC value of the strain with ceftazidime sensitivity recovery was not changed by continuous blank culture for 10 generations.The results showed that after continuous treatment of 1/4 MIC Chaiqintonglin Tablet,the sensitivity of some clinical Escherichia coli to ceftazidime could be restored,and the recovery of drug resistance of Escherichia coli had stable genetic characteristicsThe effect of Chaiqintonglin on β-lactamase production was determined by double paper.The results showed that all generations of clinically resistant Escherichia coli E4B5 after continuous induction of 1/4MIC CQTL had changed from the original ESBLs producing strain to non-ESBLs producing strain in the second generation.It is suggested that 1/4MIC Chaiqin Tonglin Tablet may inhibit ESBLs.After continuous induction of each generation of clinically resistant Escherichia coli E5A4 by 1/4MIC concentration CQTL,the first,second and third generation of strains changed from the original ESBLs producing strains to non-ESBLs producing strains.However,the bacteria of the fourth and fifth generations could gradually recover their enzymeproducing ability under the action of drug pressure.The results suggested that Chaiqin Tonglin could inhibit the β-lactamase in some extent.2.Antibacterial effect of Chaiqin Tonglin Tablet on Escherichia coli in biofilm stateThe results of the modified microplate method showed that the 1/2 MIC and1/4MIC Chaiqin Tonglin had certain inhibitory effects on pathogenic Escherichia coli in biofilm state at mature and formative stages.The 1/2 MIC-1/8 MIC-Chaiqintonglin significantly reduced the aggregation capacity of E4B5 strain.It was speculated that Chaiqintonglin tablets could affect the membrane formation capacity of biofilm by reducing the aggregation capacity of Escherichia coli.3.Effect of Chaiqin Tonglin Tablet on the adhesion and invasion ability of pathogenic Escherichia coli in urinary tractChai Qin tong Lin piece in 30 to 15 mg/ml concentration can significantly inhibit the urinary tract pathogenic e.coli adhesion ability of T24 cells,in the 30-7.5 mg/ml concentration are different degree of inhibition of urinary tract pathogenic e.coli invade T24 cells,in just 30 mg/ml concentration can inhibit T24 cell intracellular relative amounts of pathogenic e.coli,show the ability to remove intracellular bacteria.Laser confocal results showed that Chaiqintonglin tablets could reduce the invasion of Escherichia coli.The results of swimming showed that 30mg/ml and 15mg/ml Chaiqintonglin tablets could significantly inhibit the locomotion ability of Escherichia coli.Q-PCR results showed that flagella-related genes FLHD,FLHC and FLIA were significantly down-regulated by 30mg/ml Chaiqintonglin tablets.The results showed that Chaiqin Tonglin Tablet could protect the bladder epithelial cells,reduce the colonization and invasion of pathogenic Escherichia coli E4B5,and reduce the expression of flagellum gene of pathogenic Escherichia coli.