连花清瘟胶囊抗金黄色葡萄球菌生物膜研究

目的: 观察连花清瘟胶囊对成熟金黄色葡萄球菌生物膜(Staphylococcus aureau biofilm,S.a BF)的抑制作用。 方法: 采用96孔板微量稀释法检测连花清瘟胶囊和青霉素最小杀菌浓度(MBC);用结晶紫(CV)和微生物活性(WST)检测法观察连花清瘟胶囊和青霉素抗S.a BF的作用;用扫描电子显微镜(SEM)和激光共聚焦显微镜(CLSM)观察药物作用后的S.a BF形态并测量厚度。 结果: 连花清瘟胶囊和青霉素对S.a的MBC分别为20 g·L^-1和2 mg·L^-1。与对照组比较,连花清瘟胶囊显著抑制S.a BF(P<0.05,P<0.01),青霉素差异不显著。SEM观察到空白对照组S.a BF较厚,细菌形态正常,密集叠加,大小一致;连花清瘟胶囊组S.a BF由散在菌落组成,大小不一致;青霉素组S.a BF较厚,细菌形态基本正常,密集叠加,大小一致,个别细菌塌陷萎缩;CLSM观察到空白对照组S.a BF以活菌为主体,其间散落死菌;连花清瘟胶囊组S.a BF有大量死菌,和活菌相互参杂;青霉素组S.a BF以活菌为主,部分死菌分布在S.a BF表层。与空白对照组比较,连花清瘟胶囊组S.a BF厚度显著降低(P<0.01),青霉素组差异不显著,连花清瘟胶囊组S.a BF厚度显著低于青霉素组(P<0.01)。 结论: 连花清瘟胶囊具有显著的抗S.a BF作用,抗S.a BF评估体系可应用于传统中药抗菌作用研究。

Study on Lianhua Qingwen Capsule Resisting Staphylococcus aureus Biofilm

Objective: To observe the inhibition of Lianhua Qingwen capsule(LHQW) on Staphylococcus aureus biofilm(S.a BF). Method: Microdilution method was used to assay minimal bactericidal concentration (MBC) of LHQW and penicillin. The crystal violet(CV) and microbial activity (WST) assay were used to evaluate the inhibition of LHQW and penicillin on S.a BF. The scanning electron microscopy (SEM) and laser scanning confocal microscopy (CLSM) were used for observing S.a BF morphology and thickness measurement. Result: The MBC values of LHQW and penicillin against free S.a were 20 g·L^-1 and 2 mg·L^-1. Compared with the control group (control), LHQW significantly inhabited S.a BF(P<0.05,P<0.01),penicillin was no significant difference;S.a BF of the control group showed the thicker,normal bacterial form, dense overlay and the same size under the SEM, while S.a BF of LHQW group showed scattered colonies, inconsistent size, and S.a BF of penicillin group showed thicker, basically the same size, and the individual bacteria were collapse;S.a BF of control group were made of viable cells and a little dead bacteria under the CLSM, while S.a BF of LHQW group had a large number of dead bacteria, with which viable cells mixed;S.a BF of penicillin group was made of viable cells and the dead bacteria are distributed over the surface of S.a BF. Compared with the blank control group, S.a BF of LHQW group was thick significantly (P<0.01),penicillin had no significant difference;Sa BF of LHQW group was less thick significantly than the penicillin group (P<0.01). Conclusion: LHQW has significant anti- S.a BF effect. Anti-S.a BF assessment system can be used in the antibacterial effect of traditional Chinese medicine.