Fig. 8

ML385 inhibits the effects of protodioscin on the structure and morphology of lung tissues as well as the levels of lung fibrosis markers in mice with pulmonary fibrosis. A mouse lung fibrosis model was generated through tracheal injections of bleomycin, and 30 mg/kg of ML385 was injected intraperitoneally 3 h prior to the administration of high-dose protodioscin. Its effect on the therapeutic effect of protodioscin was then observed (A). Changes in lung tissue structures and extracellular matrix were visualized by HE and Masson staining (magnification × 100) (B, C) before analyzing the degree of pulmonary fibrosis in each group of mice with the Ashcroft score (G). The hydroxyproline contents of lung tissues were determined by biochemical methods (F). The positive expression areas of col-I and α-SMA in lung tissues were analyzed by immunohistochemical methods (magnification × 100) (D, E). Determining the mRNA expression levels of col1a1, FN1, CTGF and α-SMA in lung tissues was undertaken through qRT-PCR (H–K), while the protein levels of col-I, FN1, CTGF and α-SMA were quantified by WB (L). ML385 indicates the group treated with ML385 prior to high-dose treatment; BLM + H indicates the high-dose treatment group; BLM indicates the lung fibrosis model group; Control indicates the control group. Expression of data was undertaken in form of mean ± standard deviation. The sample size, particularly per group, was eight, * P < 0.05; *It is suggestive of statistical significance at the P threshold below 0.05; **Denotes heightened significance at the P threshold below 0.01; ***Reflects substantial significance at the P threshold below 0.001; ****Demonstrates remarkable significance at the P threshold below 0.0001