Antibacterial active compounds from Hypericum ascyron L. induce bacterial cell death through apoptosis pathway.
Hypericum ascyron L. has been used as a traditional medicine for the treatment of wounds, swelling, headache, nausea and abscesses in China for thousands of years. However, modern pharmacological studies are still necessary to provide a scientific basis to substantiate their traditional use. In this study, the mechanism underlying the antimicrobial effect of the antibacterial activity compounds from H. ascyron L. was investigated. Bioguided fractionation of the extract from H. ascyron L. afforded antibacterial activity fraction 8. The results of cup plate analysis and MTT assay showed that the MIC and MBC of fraction 8 is 5 mg/mL. Furthermore, using Annexin V-FITC/PI, TUNEL labeling and DNA gel electrophoresis, we found that cell death with apoptosis features similar to those in eucaryon could be induced in bacteria strains after exposure to the antibacterial activity compounds from H. ascyron L. at moderate concentration. In addition, we further found fraction 8 could disrupt the cell membrane potential indicate that fraction 8 exerts pro-apoptotic effects through a membrane-mediated apoptosis pathway. Finally, quercetin and kaempferol 3-O-β-(2″-acetyl)-galactopyranoside, were identified from fraction 8 by means of Mass spectrometry and Nuclear magnetic resonance. To our best knowledge, this study is the first to show that Kaempferol 3-O-β-(2″-acetyl)-galactopyranoside coupled with quercetin had significant antibacterial activity via apoptosis pathway, and it is also the first report that Kaempferol 3-O-β-(2″-acetyl)-galactopyranoside was found in clusiacea. Our data might provide a rational base for the use of H. ascyron L. in clinical, and throw light on the development of novel antibacterial drugs.
Key Lab of Industrial Fermentation Microbiology of the Ministry of Education, Tianjin Key Lab of Industrial Microbiology, College of Biotechnology, Tianjin University of Science and Technology, Tianjin, 300457, PR China.
Xiu-Mei
XM
Li
Key Lab of Industrial Fermentation Microbiology of the Ministry of Education, Tianjin Key Lab of Industrial Microbiology, College of Biotechnology, Tianjin University of Science and Technology, Tianjin, 300457, PR China. Electronic address: luoxuegang@hotmail.com.
Xue-Gang
XG
Luo
Tianjin Key Laboratory of Pulp & Paper, Tianjin University of Science & Technology, Tianjin, 300457, PR China; State Key Laboratory of Tree Genetics and Breeding, Northeast Forestry University, Harbin, 150040, PR China.
Chuan-Ling
CL
Si
Key Lab of Industrial Fermentation Microbiology of the Ministry of Education, Tianjin Key Lab of Industrial Microbiology, College of Biotechnology, Tianjin University of Science and Technology, Tianjin, 300457, PR China.
Nan
N
Wang
Key Lab of Industrial Fermentation Microbiology of the Ministry of Education, Tianjin Key Lab of Industrial Microbiology, College of Biotechnology, Tianjin University of Science and Technology, Tianjin, 300457, PR China.
Hao
H
Zhou
Key Lab of Industrial Fermentation Microbiology of the Ministry of Education, Tianjin Key Lab of Industrial Microbiology, College of Biotechnology, Tianjin University of Science and Technology, Tianjin, 300457, PR China.
Jun-Fang
JF
He
Key Lab of Industrial Fermentation Microbiology of the Ministry of Education, Tianjin Key Lab of Industrial Microbiology, College of Biotechnology, Tianjin University of Science and Technology, Tianjin, 300457, PR China; Institute of Biology and Medicine, Wuhan University of Science and Technology, Wuhan, 430000, PR China. Electronic address: tony@tust.edu.cn.
Tong-Cun
TC
Zhang
D000900:Anti-Bacterial Agents / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects*; D004305:Dose-Response Relationship, Drug; D016972:Enterobacter cloacae / Q000166:cytology / Q000187:drug effects; D004926:Escherichia coli / Q000166:cytology / Q000187:drug effects; D020902:Hypericum / Q000737:chemistry*; D007711:Klebsiella pneumoniae / Q000166:cytology / Q000187:drug effects; D008826:Microbial Sensitivity Tests; D050296:Microbial Viability / Q000187:drug effects*; D016982:Micrococcus luteus / Q000166:cytology / Q000187:drug effects; D015394:Molecular Structure; D010936:Plant Extracts / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology*; D013211:Staphylococcus aureus / Q000166:cytology / Q000187:drug effects; D013329:Structure-Activity Relationship
D000900:Anti-Bacterial Agents; D010936:Plant Extracts
Antimicrobial activity; Apoptosis; Hypericum ascyron L.; Preparative high performance liquid chromatography; Structure identification; Structure–activity relationship