Bleomycin blm

Wild-type (C57BL/6J) mice were purchased from the Model Animal Research Center of Nanjing University. Mutyh^+/− mice were previously established and backcrossed to C57BL/6J mice as described previously [15 (link)], and Mutyh^−/− mice were obtained via inbreeding. Eight-week-old male mice (18-22 g) were used in the present experiment. All animals were maintained under standard conditions of 50% relative humidity, 21 ± 2°C, and a 12 h light cycle. To induce pulmonary fibrosis, 4 mg/kg bleomycin (BLM, Nippon Kayaku Co., Tokyo, Japan) was instilled intratracheally in a single dose, and an equal volume of normal saline (NS) was used as a control. The experimental mice were euthanized on days 7 (D7), 14 (D14), and 28 (D28) after intratracheal instillation. Lung tissues and serum from wild-type and Mutyh^−/− mice were collected for further analysis (Figure 1(a)). The animal experiments were performed based on the Jiangsu Provincial Experimental Animal Manage Committee under Contract 2011-0069, and the Experimental Animal Ethics Committee of Nanjing University approved the experimental protocols.

For inducing skin fibrosis, the back skins of 8-week-old female BALB/c mice were shaved, and for 21 days, the mice were daily subcutaneously injected with 100 μg/100 μL bleomycin (BLM) (Nippon Kayaku, Tokyo, Japan) in sterile saline [33 (link)] (Additional file 1). The mASCs activated with LMWH (hep-mASCs) were cultured in DMEM/F-12 containing 10% FBS and 1.0% Pen-Strep supplemented with LMWH (10 and 100 μg/mL). In this study, to assess the effect of hep-mASCs, mice were assigned to the following groups, with n = 9 in each group: BLM-induced SSc (BLM-alone), BLM-induced SSc administered mASCs (BLM-mASC), BLM-induced SSc administered mASCs activated with 10 μg/mL LMWH (BLM-hep10-mASC), and BLM-induced SSc administered mASCs activated with 100 μg/mL LMWH (BLM-hep100-mASC). The mASCs cultured with or without LMWH were intravenously injected into the tail vein with 100 μL of PBS. In the BLM-alone group, 100 μL of PBS was intravenously injected into the tail vein. The number of administered cells was 2.5 × 10⁴ cells. All three treatments were performed on day 7 following BLM administration. The mice were euthanised and their skins were harvested at 21 days after administering BLM.