[Mechanism study on Bushen Yisui Shengxue Formula regulating Hippo/Yap pathway to restore bone marrow hematopoietic...

This study aimed to investigate the therapeutic effect of Bushen Yisui Shengxue Formula on aplastic anemia(AA) and to explore the mechanism by which this prescription alleviates bone marrow failure in AA mice through modulation of the Hippo/Yes-associated protein(Yap) pathway. BALB/c mice were randomly assigned to control, model, and Bushen Yisui Shengxue Formula(traditional Chinese medicine) groups. The AA model was established by X-ray irradiation combined with DBA/2 mouse lymphocyte infusion. After successful modeling, the traditional Chinese medicine group received Bushen Yisui Shengxue Formula via gavage for 14 consecutive days, while the model and control groups received normal saline. Following the intervention, general conditions such as mental status, activity, and body weight were observed in each group. Blood samples were collected from the orbital vein for peripheral blood cell counts. Bone marrow pathology was examined by hematoxylin-eosin(HE) staining, while Wright-Giemsa staining was used to assess the number, classification, and morphology of bone marrow cells. Bone marrow nucleated cells were counted under a microscope, and apoptosis levels were measured by flow cytometry. The mRNA expression of Hippo/Yap pathway: Yap, transcriptional enhanced associate domain(TEAD), and telomerase reverse transcriptase(TERT) was detected using quantitative polymerase chain reaction(qPCR), and their protein expression levels were evaluated by Western blot. The results showed that, compared with the control group, mice in the model group exhibited weight loss, weakness, and lethargy; reduced peripheral blood cell counts; decreased bone marrow hematopoietic cells with increased adipocyte infiltration; elevated apoptosis; lower bone marrow nucleated cell counts; and downregulated mRNA and protein expression of TERT, Yap, and TEAD. In contrast, Bushen Yisui Shengxue Formula treatment significantly restored body weight, mental state, and physical activity in the AA mice. It also increased peripheral blood WBC, RBC, and PLT counts; enhanced bone marrow hematopoiesis while reducing fat infiltration; decreased apoptosis; raised bone marrow nucleated cell counts; and upregulated both mRNA and protein levels of TERT, Yap, and TEAD compared with the model group. These findings suggest that Bushen Yisui Shengxue Formula can ameliorate bone marrow failure in AA by modulating the Hippo/Yap pathway, which is implicated in AA pathogenesis, indicating its potential as a promising therapeutic agent for AA treatment.