Experiment Of Reversal On Drug-Resistance Of Escherichia Coli Of Beta-Lactam Aantibiotics By Chinese Traditional Medicine

OBJECTIVE To study the reversal effect of three kinds of traditional Chinese medicine (San Huang Tang, Coptidis Decoction for Detoxification and Wu Wei Xiao Du Yin) to bacterial drag resistance of E.coli which generates extended-spectrum-lactamases, and approach the optimal culture time and the optimal drag concentration of these Chinese medicine that form the best reversal effect. And to study the enzyme inhibition effect of three kinds of traditional Chinese medicine by determining activity and content of extended spectrum lactamases (ESBLs), and approach the reversal mechanism of bacterial drug resistance of E.coli which generates extended-spectrum-lactamases.METHOD E.coli which generates extended-spectrum-lactamases is screening by disk diffusion method and double-disk diffusion test. Minimum Inhibiory Concentration is determined by tube double dilution method. E.coli is cultured in under MIC and the reversal effect is compared in different culture time and different culture concentration under MIC. And the cultural time and concentration of optimal reversal effect is selected. Meanwhile ESBLs is extracted and it's activity and content is determined, the reversal mechanism can be explored.RESULT Drag resistance E.coli was collected from a provincial hospital and E.coli which generates extended-spectrum-lactamases was screening by disk diffusion method and double-disk diffusion test. The E.coli was experimental sample. Minimum Inhibiory Concentration (MIC) was determined by E.coli ATCC25922. And the MIC(San Huang Tang) was 0.5g/ml, the MIC(Coptidis Decoction for Detoxification) was 0.6g/ml, and the MIC(Wu Wei Xiao Du Yin) was 0.7g/ml. The experimental sample was cultured in clavulanic acid and the minimum reversal concentration can make the resistance to become sensitive. The experiment indicated that the minimum reversal concentration of clavulanic acid was 15μg/ml. The E.coli under this culture condition was positive control. The resistance E.coli was cultured in under MIC and the reversal effect was compared in different culture time and different culture concentration under MIC. The data was manipulated by SPSS11.5 and the optimal reversal result was 0.2g/ml 24h(San Huang Tan), 0.4g/ml 12h(Coptidis Decoction for Detoxification), 0.3g/ml 36h and 0.4g/ml 48h(Wu Wei Xiao Du Yin), but there was the significant difference(P<0.05) between the optimal reversal result and positive control. The extended-spectrum-lactamases (ESBLs) was extracted and three dimensional test indicated that hypersound quassation had been better than rep-freeze thawing. The optical density of cefotaxim and ESBLs was determined and activity of enzyme was calculated by formula. The result indicated that Chinese medicinal formulae could degrade drug resistance by inhibiting activity of enzyme. There was no significant difference (P>0.05) between the reversal result and positive control and there was the significant difference (P<0.05) between the reversal result and negative control. There was no significant difference (P>0.05) between each Chinese medicinal formulae. The every prescription of San Huang Tang could inhibit activity of enzyme, but there was the significant difference (P<0.05) between the reversal result and positive control and there was the significant difference (P<0.05) the reversal result and negative control, also. There was no significant difference (P>0.05) between each prescription of San Huang Tang. The content of ESBLs was calculated by standard curve. The results indicated that Chinese medicinal formulae could also inhibit express of ESBLs. There was the significant difference (P<0.05) between the reversal result and positive control, but there was the significant difference (P<0.05) the reversal result and negative control. The express of ESBLs was inhibited by every prescription. There was no significant difference (P>0.05) between the reversal result and positive control, but there was the significant difference (P<0.05) the reversal result and negative control.CONCLUSION The experiment further confirm that Traditional Chinese medicine have the reversal effect to bacterial drug resistance which is relate with the concentration of traditional Chinese medicine and the culture time. And San Huang Tang is the best reversal effect of the three Traditional Chinese medicine. One of the major route is inhibiting the activity and express of ESBLs. But there may be otherreversal mechanisms is need further investigated...