双黄连、清开灵对耐药大肠埃希菌R质粒及β-内酰胺酶的影响

Influences of Shuanghuanglian Injectable Powder and Qingkailing Injection on R plasmids and β-lactamase of drug-resistant Escherichia coli

目的探讨清热解毒中药双黄连、清开灵对产超广谱β-内酰胺酶(ESBLs)大肠埃希菌R质粒的消除作用及对β-内酰胺酶活性的影响。方法取分离于临床的ESBLs阳性大肠埃希菌,用质粒提取试剂盒提取质粒,应用琼脂糖凝胶电泳、紫外凝胶成像仪分析中药作用前后质粒消除情况;以超声破碎法获取β-内酰胺酶提取液,用Bradford法进行蛋白定量,以头孢硝噻吩(Nitrocefin)为底物测定β-内酰胺酶活性。结果对双黄连、清开灵作用前后质粒图谱比较发现经中药处理后均丢失了一条质粒带;肉汤菌、经舒巴坦、双黄连以及清开灵作用后细菌β-内酰胺酶活性分别为(13.43±0.40)、(3.49±0.12)、(3.54±0.27)、(3.35±0.12)U/mg,与肉汤菌相比,经双黄连、清开灵作用后酶活性降低,差异具有统计学意义(P<0.01);其作用与舒巴坦相当。结论双黄连及清开灵具有一定消除耐药大肠埃希菌质粒的作用,并且可抑制其β-内酰胺酶活性。

Objective To discuss the elimination effect of Shuanghuanglian Injectable Powder and Qingkailing Injection on R plasmids of Escherichia coli （E. coli ） producing super broad-spectrum β-1actamase （ESBLs） and influence on the activity of ESBLs. Methods E. coli with ESBLs positive were colleted from clinic and R plasmids were extracted with plasmid kit. The elimination of R plasmids was analyzed by using agarose gel electrophoresis and UV gel imager before and after the administration of Chinese medicinal. The extracting solution of ESBLs was collected by using ultrasonication and protein quantified by using Bradford method. The activity of ESBLs was determined taking Nitrocefin as substrate. Results Before and after the administration of Shuanghuanglian Injectable Powder and Qingkailing Injection, the comparison of plasmid maps showed that a plasmid band was lost. The activity of ESBLs was, respectively, （13.430.40）U/mg, （3.490.12）U/mg, （3.540.27）U/mg and （3.350.12）U/mg in broth group, Sulbactam group, Shuanghuanglian group and Qingkailing group. The activity of ESBIs decreased in Shuanghuanglian group and Qingkailing group compared with broth group （ P 〈 0. 01 ） and was similar to that in Sulbactam group. Conclusion Shuanghuanglian Injectable Powder and Qingkailing Injection have some elimination effects on R plasmids of drug-resistant E. coli and can inhibit the activity of ESBLs.