Inhibitory effects of(R)-(+)-pulegone on drug-resistant Mycobacterium tuberculosis and itsbiofilm-regulating genes
Objective This study investigated in vitro inhibitory effects of(R)-(+)-pulegone on drug-re-sistant Mycobacterium tuberculosis and its influence on biofilm-regulating genes Rv0024,Rv2872and Ra1362.Methods Drug-resistant Mycobacterium tuberculosis strains containing the biofilm-regulating genes Rv0024,Rv2872 and Ra1362 were screened using polymerase chain reaction(PCR).The minimum inhibitory concentration(MIC)and minimum bactericidal concentration(MBC)of(R)-(+)-pulegone a-gainst biofilm-mediated drug-resistant Mycobacterium tuberculosis were determined using the methylene blue dye method and liquid culture counting method,respectively.Reverse transcription-polymerase chain reaction(RT-PCR)was employed to assess the impact of(R)-(+)-pulegone on the expression of biofilm-regulating genes Rv0024,Rv2872 and Ra1362.Results The MIC and MBC of(R)-(+)-pulegone against drug-resistant Mycobacterium tuberculosis were found to be 14.79 mg/mL and 29.59 mg/mL,respective-ly,using the methylene blue dye method at a bacterial density of 3×107 CFU/mL and 3×106 CFU/mL.The MIC90 and MBC of(R)-(+)-pulegone determined by the liquid culture counting method were 19.89 mg/mL and 24.62 mg/mL,respectively.The results of RT-PCR showed that long leaved menthone could inhibit the expression of Rv0024 and Ra1362,and promote the expression of Rv2872(P<0.05).Conclusions(R)-(+)-pulegone inhibits the growth of drug-resistant Mycobacterium tuberculosis by regu-lating the expression of the main genes Rv0024,Rv2872and Ra1362 in the biofilm of drug-resistant Myco-bacterium tuberculosis.
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