三黄汤对E.coli内ESBLs基因表达的干预性研究-《西部中医药》

文章信息/Info

Title:: Intervention Study of SanHuangTang on Gene Expression of ESBLs within E.coli

Author(s):: JIA Yun-peng1, CHENG Ning2; 1 Gansu Province Hospital of Traditional Chinese Medicine, Lanzhou 730050, China; 2 Center of Reproductive Health and Birth Defects in Lanzhou University

摘要:: 目的：观察三黄汤水煎剂对E.coli细胞内编码ESBLs基因TEM,CTX-M,SHV,OXA以及编码膜外排泵蛋白AcrAB-TolC基因Acr-AB表达的干预效果，探讨中药对细菌耐药性逆转的可能性。方法：采集鉴定ESBLs阳性E.coli菌株30株，用试管二倍稀释法确定三黄汤的MIC（最低抑菌浓度），选取低于MIC的不同浓度在规定时间下培养耐药E.coli，测定不同浓度逆转效果和最佳逆转浓度，并以最佳逆转浓度培养24小时后提取菌体的总RNA进行RT-PCR，检测TEM,CTX-M,SHV,OXA，Acr-AB的表达量是否受到影响。结果：三黄汤水煎剂对ESBLs阳性大肠埃希菌最低抑菌浓度为0.2 g/mL；对照组与用药组相比，OXAQ差异显著，有统计学意义(P＜0.05)，而OXAJ无统计学意义(P＞0.05)，5个基因中，编码OXA型ESBLs的基因表达量在干预前后有统计学意义(P＜0.05)。结论：中药汤剂能通过抑制ESBLs阳性E.coli内ESBLs的活性和表达量，降低细菌耐药性，且其逆转作用与中药浓度有关；0.2 g/mL三黄汤对ESBLs阳性E.coli体内的ESBLs编码基因OXA有抑制作用，可使其表达量下降；尚未发现三黄汤对受试菌TEM,CTX-M,SHV,Acr-AB基因有明确的干预作用。

Abstract:: Objective: To explore the possibility of TCM on reversing of drug resistance in bacteria by observing intervention of SanHuangTang on gene expression which encoding TEM, CTX-M, SHV, OXA and encoding membrane efflux pump AcrAB-TolC protein in ESBLs positive E.coli. Method: All 30 strains of E.coli with positive ESBLs were collected and identified, cultured within limited time in different concentration below minimum inhibitory concentration(MIC), which was fixed with tube double dilution method. Reversal effects under different concentrations and optimal reversal concentration were detected and then total RNA was extracted with RT-PCR after incubation 24 hours to detect whether expressions of TEM, CTX-M, SHV, OXA and Acr-AB were affected or not. Result: SanHuangTang could inhibit activity of ESBLs or reduce the production through intervention on expression of gene OXA therefore to reverse drug resistance of bacteria. The reversal effect is related with the concentration of herbs. Conclusion: Decoction of herbs can reduce drug resistance of bacteria by inhibiting activity and expression of ESBLs in positive E.coli; 0.2 g/mL SanHuangTang can inhibit OXA which encoding ESBLs in ESBLs-positive E.coli to decrease its expression; there is no evidence showing clear intervention of SanHuangTang on expression of TEM, CTX-M, SHV, Acr-AB.