Male C57/BL6 mice 8 to 12 weeks old (20–25 g, SLC, Shizuoka, Japan) were anesthetized with intraperitoneal ketamine (80 mg/kg) and xylazine (10 mg/kg) and received a single tracheal injection of 2 mg/kg bleomycin sulfate (Nippon Kayaku, Tokyo, Japan) in 50 μL sterile saline using a Microsprayer Aerosolizer (PennCentury, Philadelphia, PA, USA) on day 0. All mice were killed by cervical dislocation and lungs were harvested at day 7, 14, 21 or 28 after intratracheal bleomycin or saline treatment. This study was approved by the Animal Care and Use Committee of Hamamatsu University School of Medicine and all experiments were performed according to guidelines of this Committee.
Bleomycin sulfate
Manufactured by Nippon Kayaku
Sourced in Japan
Bleomycin sulfate is a pharmaceutical compound used as a laboratory reagent. It functions as an antitumor antibiotic, inhibiting DNA synthesis.
Automatically generated - may contain errors
Lab products found in correlation
Microsprayer aerosolizer, by Penn-Century (2 mentions)
Midazolam, by Sandoz (1 mentions)
Vetorphale, by Meiji Seika Pharma (1 mentions)
Medetomidine, by Nippon Zenyaku Kogyo (1 mentions)
Atipamezole, by Nippon Zenyaku Kogyo (1 mentions)
Recombinant human tgf β1, by Thermo Fisher Scientific (1 mentions)
Penicillin, by Beyotime (1 mentions)
Streptomycin, by Beyotime (1 mentions)
5 protocols using bleomycin sulfate
1
Bleomycin-Induced Lung Fibrosis in Mice
2
Bleomycin-Induced Lung Injury Model
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Male C57BL/6J mice aged 8–10 weeks were intratracheally instilled with 80 μl of saline containing 3.2 mg/kg body weight of bleomycin sulfate (Nippon Kayaku Co., Ltd., UK) to induce BLM-ILD or 80 μl of saline alone as the control. Mice were anesthetized with an intraperitoneal injection of 0.75 mg/kg of medetomidine (Nippon Zenyaku Kogyo, Fukushima, Japan), 4.0 mg/kg of midazolam (Sandoz, Tokyo, Japan), and 5.0 mg/kg of vetorphale (Meiji Seika Pharma, Tokyo, Japan) before the procedure, and medetomidine was antagonized by a peritoneal injection of 0.75 mg/kg of atipamezole (Nippon Zenyaku Kogyo, Fukushima, Japan) after the procedure. The lungs were dissected under the anesthesia described above before or on day 1, 3, 7, 10, or 14 after the administration of BLM.
3
Evaluating TGF-β1 and Bleomycin Effects
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Dulbecco’s modified eagle medium (DMEM) and fetal bovine serum (FBS) were purchased from Hyclone (UT, USA). Penicillin and streptomycin were purchased from Beyotime (Jiangsu, China). Human recombinant TGF-β1 was provided from Peprotech (Rocky Hill, NJ). Bleomycin sulfate was supplied by Nippon Kayaku Co., LTD. (Japan). Information including source, catalog, dilution ratio, and storage conditions of primary/secondary antibodies are presented in Additional file 1 : Table S1, and the structural information and source of the active compounds are listed in Additional file 1 : Table S2.
4
Bleomycin-Induced Lung Injury in Mice
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C57/BL6 male mice 9 to 12 weeks old (20–25 g, SLC, Shizuoka, Japan) were anesthetized with intraperitoneal ketamine (80 mg/kg) and xylazine (10 mg/kg). Using a microsprayer (Microsprayer Aerosolizer; PennCentury, Philadelphia, PA, USA), mice were instilled a single tracheal injection of 2 mg/kg bleomycin sulfate (Nippon Kayaku, Tokyo, Japan) in 50 μL sterile saline on day 0. Control mice received tracheal injection of 50 μL sterile saline. All mice were killed by cervical dislocation and lungs were harvested at day 7 after intratracheal saline or bleomycin injection. This study was approved by the Animal Care and Use Committee of Hamamatsu University School of Medicine and all experiments were performed according to guidelines of this Committee.
5
Botanical Extraction and Chemical Analysis
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Bleomycin sulfate was purchased from Nippon Kayaku Co. (Tokyo, Japan). Ketamine hydrochloride was received from Gedeon Richter, Hungary. Sodium thiopental was gifted by Rhone Poulenc Rorer, France. Sodium tungsten was purchased from BDH, England. Other reagents were received from Merck Co, Germany. All other reagents were highly commercially available. Chamomile dried flowers also were obtained from M. chamomilla plant, Compositae family, GolDaru company, Isfahan, Iran.
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