加味补阳还五汤促进肝细胞生长因子旁分泌防治术后腹腔粘连的实验研究OA

Objective To investigate the mechanism of Modified Buyang Huanwu Decoction(MBHD)by which regulates hepatocyte growth factor(HGF),inhibits the transforming growth factor-β1(TGF-β1)/Smad3 signaling pathway,reduces epithelial-mesenchymal transition(EMT)in peritoneal mesothelial cells(PMCs),and thereby prevents and treats postoperative abdominal adhesions(PAA).Methods Sixty male Sprague-Dawley rats were randomly divided into four groups using a random number table(n=15/group):normal,model,Smad3 inhibitor(5×10-3 g/kg),and MBHD(21 g/kg).The model and normal groups received an equal volume of normal saline by gavage.A PAA model was established in all rats using a multifactorial injury method,except for the normal group.All rats were euthanized on postoperative day 28 for sample collection.Histopathological changes in adhesion tissues were evaluated using hematoxylin-eosin and Masson's trichrome staining.Immunohistochemistry was used to detect the expression levels of HGF,E-cadherin(E-cad),and α-smooth muscle actin(α-SMA)in omental adipose tissue and cecal serosal tissue.The mRNA expression levels of TGF-β1 and Smad3 in adhesion tissues were determined using quantitative reverse transcriptase PCR.Protein expression levels of HGF,E-cad,α-SMA,TGF-β1,and phosphorylated Smad3(p-Smad3)were analyzed using western blotting.Results Compared with the normal group,rats in the model group showed disordered and loose arrangement of PMCs in the cecal serosal layer,obvious fibrous connective tissue hyperplasia and inflammatory cell infiltration,and increased collagen fiber deposition.Compared with the model group,the MBHD group and Smad3 inhibitor group showed an orderly and compact arrangement of PMCs in the serosal layer,with only a small amount of local fibrous connective tissue hyperplasia.Inflammatory cell infiltration was significantly reduced,collagen fiber proliferation was markedly decreased,and the PMCs were arranged compactly.The expression levels of HGF in omental adipose tissue,as well as HGF and E-cad in cecal serosal tissue,were increased(P<0.05),whereas the expression levels of TGF-β1,p-Smad3,and α-SMA in cecal serosal tissue were decreased(P<0.01,P<0.05).Conclusion MBHD may prevent and treat PAA by promoting peritoneal paracrine secretion of HGF,inhibiting the TGF-β1/Smad3 signaling pathway,and reducing EMT of PMCs.