A Study On The Inhibitory Effect Of Artesunate On Ureaplasma Urealyticum In Vitro

Mycoplasma is the smallest prokaryotic microorganism that can be cultured in inanimate medium.It mainly exists in oral cavity,respiratory tract and genitourinary tract in human body and is related to many diseases.In recent years,with the widespread use of antibiotics,the drug resistance of mycoplasma is becoming more and more serious.Screening,studying and developing new anti-mycoplasma drugs from traditional Chinese medicine to give full play to the role of traditional Chinese medicine in the prevention and treatment of diseases is not only a positive response to the national strategy of"healthy China",but also has great feasibility and good application value.Objective:To find out the epidemiological characteristics and drug sensitivity of Mycoplasma infection in urogenital tract in Changsha,which will provide a basis for clinical treatment.To study the inhibition effect of artesunate,Plantago asiatica L.,Lysimachia christinae Hance and Pyrrosia lingua（Thunb.）Farwell on Ureaplasma urealyticum in vitro,and to find out their minimum inhibitory concentration.To investigate the inhibition effect of artesunate combined with roxithromycin and levofloxacin on UU in vitro.Methods:In order to find out the relationship between the infection of UU and Mycoplasma hominis on the ages,genders,and the changes in different years,the culture results of UU and MH in a hospital of Changsha from2016 to 2020 were retroactively analyzed according to gender,age and time groups.Based on the drug sensitivity data of mycoplasma from 2016to 2020,we may understand the sensitivity of UU and MH to minocycline,doxycycline,josamycin,erythromycin,azithromycin,clarithromycin,sparfloxacin,ofloxacin and levofloxacin in Changsha area and their changes in different years;Quantitative Real-time PCR was used to detect the concentration of UU,and to find out the relationship between CFU and copies of UU.The minimum inhibitory concentrations of artesunate,Plantago asiatica L.,Lysimachia christinae Hance and Pyrrosia lingua（Thunb.）Farwell against UU were determined by minimal broth dilution method.The inhibition effect of artesunate combined with roxithromycin or levofloxacin on UU in vitro was detected by checkerboard dilution method.Results:1.A total of 6668 positive mycoplasma cases were detected in13833 samples,The positive rate was higher in females than in males（58.41%vs 25.35%,χ2=1292.6,P<0.01）and the risk of positive mycoplasma in females was 4.191 times than that in males（P<0.001）.From 2016 to 2020,the annual positive rate varied significantly only infemales and gradually went up（P<0.05）.The detection rates of mycoplasma in both males and females decreased along with age.Meanwhile,the detection rates of mycoplasma in females were lower in the age group of 41-and 51-compared to other age groups.Among 6668positive mycoplasma cases,5982 were positive for UU（89.71%）,165 for MH（2.47%）and 521 for both（7.82%）,certificating that UU is the main type of mycoplasma infection in urogenital tract of patients in Changsha.Semi-quantitative analysis showed that carrying rates for both UU and MH were high.Drug sensitivity test showed that josamycin,doxycycline and minocycline could be used for clinical treatment of urogenital mycoplasma infection.The use of quinolone antibiotics,such as sparfloxacin,ofloxacin,levofloxacin,required caution.2.Artesunate had a strong inhibitory effect on UU in vitro,with MIC₅₀≤0.5 mg/m L and MIC₉₀≤1 mg/m L;Lysimachia christinae Hance and Pyrrosia lingua（Thunb.）Farwell had a less inhibitory effect on UU,for Lysimachia christinae Hance,the MIC₅₀≤4 mg/m L and MIC₉₀≤8mg/m L;for Pyrrosia lingua（Thunb.）Farwell,the MIC₅₀≤4 mg/m L and MIC₉₀≤8 mg/m L;However,the inhibitory effect of Plantago Fructus on UU in vitro was weak,with MIC₅₀≤16 mg/m L and MIC₉₀>16 mg/m L.3.The partly inhibition effect of artesunate combined with roxithromycin on UU was synergistic and partly synergistic,the ratio of FICI<1 was 31.42%;The partly inhibition effect of artesunate combined with levofloxacin on UU was also synergistic and partly synergistic,the ratio of FICI<1 was 33.33%.Conclusions:1.There are many influencing factors which associated with detection of urogenital mycoplasma,and further determination is needed to demonstrate the correlation between mycoplasma and urogenital tract disease.Our data suggests that routine screening and treatment of UU and MH in the urogenital tract is not recommended.2.Tetracycline drugs can be used in the clinical treatment of mycoplasma infection in the urogenital tract,but the use of quinolones and macrolides antibiotics should be cautious.3.Artesunate,Lysimachia christinae Hance and Pyrrosia lingua（Thunb.）Farwell have obvious inhibitory effects on UU in vitro,which is worthy of further study.4.Artesunate combined with roxithromycin or levofloxacin has inhibitory effect on partial UU in vitro and has certain combined drug value.