Journal: Communications Medicine
Article Title: Repurposing caspofungin as a small-molecule inhibitor of Clostridium perfringens α-toxin for treatment of gas gangrene
doi: 10.1038/s43856-026-01503-y
Figure Lengend Snippet: A HUVECs were cultured for 4 h in the presence or absence of 500 ng/ml α-toxin and the indicated concentrations of micafungin or caspofungin. Cell viability was measured using Cell Counting Kit-8 and expressed relative to the control. B HUVECs were cultured in the presence or absence of 500 ng/ml α-toxin and 50 µM micafungin or caspofungin, and a LDH leakage assay was performed using Cytotoxicity LDH Assay Kit-WST according to the manufacturer’s protocol. LDH release was expressed relative to the control. C HUVECs were cultured for 24 h in the presence or absence of 10 ng/ml α-toxin, 10 µg/ml peptidoglycan (PGN), and the indicated concentrations of micafungin or caspofungin. Interleukin-6 (IL-6) levels in the culture medium were measured using a human IL-6 Quantikine ELISA kit according to the manufacturer’s protocol. D Bone marrow cells were cultured for 3 h in the presence or absence of 100 ng/ml α-toxin and the indicated concentrations of micafungin or caspofungin, and the cells were labeled with antibodies diluted in PBS containing 2% FBS after blocking Fc-receptors with purified rat anti-mouse CD16/CD32. To quantify the amount of CD11b, a flow cytometry analysis was performed using Guava easyCyte. The mean fluorescence intensities of CD11b in Gr-1 + cells are shown. E C57BL/6J mice were injected intraperitoneally with 600 ng of α-toxin and 300 µg of micafungin or caspofungin, which had been pre-mixed and diluted in PBS. The survival of mice was monitored, and Kaplan–Meier survival curves are shown. The total number of mice used in the experiments was 63. Median survival (hours): Control, 6; caspofungin, not reached; micafungin, 6. A one-way ANOVA ( A – D ) or the Log-rank test ( E ) was employed to assess significance. Values are the mean ± standard deviation for panels ( A – D ) ( n = 3).
Article Snippet: After the treatment of cells with α-toxin, peptidoglycan (PGN), and the test compounds, culture supernatants were harvested and interleukin-6 levels were measured using a human IL-6 Quantikine ELISA kit (R&D Systems, MN, USA).
Techniques: Cell Culture, Cell Counting, Control, Lactate Dehydrogenase Assay, Enzyme-linked Immunosorbent Assay, Labeling, Blocking Assay, Purification, Flow Cytometry, Fluorescence, Injection, Standard Deviation
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