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Objective: To explore the protective effect of Linggui Zhugan Decoction (LGZGD) medicated serum on myocardial fibrosis induced by transforming growth factor-β1 (TGF-β1). Methods: Using enzyme digestion method, combined with differential adherence to isolate and culture Sprague-Dawley (SD) suckling mouse Cardiac fibroblasys (CFB) in vitro. Divided into: blank group, blank rat serum group, model group, and LGZGD medicated serum group (5%、10%、20% ). Except for blank group and blank rat serum group, they were stimulated with 5 ng/ml TGF-β1 for 12 hours, and then then intervene with LGZGD medicated serum (5%、10%、20% ) and continue to culture for 24 hours. Use immunofluorescence and Western blot (WB) to detect the expression of α-smooth muscle actin (α-SMA), Enzymelinked immunosorbent assay (ELISA) and WB to detect type I collagen (Collagen I), type I collagen (Collagen III) and fibronectin (FN) expression. Results: Compared with the blank group, the expressions of Collagen I, Collagen III, α-SMA and FN in the model group were significantly increased (P<0.01); Compared with the model group, the expressions of Collagen I and Collagen III in each concentration group of the experiment were significantly reduced (P<0.01); the expression of α-SMA and FN were significantly reduced (P<0.01). Conclusions: LGZGD has an inhibitory effect on collagen synthesis and the expression of α-SMA and FN, indicating that the anti-fibrosis effect of LGZGD is related to it. [ABSTRACT FROM AUTHOR]