Effect Of MGP-mediated TGF-β/BMP4 Pathway On PDGF-induced Proliferation Of Human Pulmonary Artery Smooth Muscle Cells

Objective:1.Human pulmonary artery smooth muscle cells(HPASMCs)were cultured in vitro,and PDGF was used to induce proliferation to construct a pulmonary hypertension cell model.2.CCK-8 method,scratch experiment,Western Blot technique were used to investigate the regulation effect of MGP-mediated TGF-β/BMP4 pathway on PDGFinduced proliferation of human pulmonary artery smooth muscle cells.Methods:1.The proliferation-promoting effect of 20ng/ml PDGF on HPASMCs for 24 h,48h,72 h was studied by morphology,CCK8 method,scratch test and other methods.2.The expression of MGP and BMP4 in the proliferation of pulmonary artery smooth muscle cells induced by 20ng/ml PDGF for 24 h,48h and 72 h was detected by Western blot.Results:1.The cell proliferation of HPASMCs in the normal group and the PDGF group was observed by morphology.The results suggested that the intervention of PDGF could promote the proliferation of HPASMCs in a time gradient dependence.When treated with 20ng/ml PDGF for 24 h,48h and 72 h,the cell proliferation state was significantly enhanced compared with the control group at the same time point.2.CCK-8 method was used to detect the effect of PDGF on HPASMCs proliferation.The results showed that compared with the control group,the proliferation of human pulmonary artery smooth muscle cells in the experimental group treated with 20ng/ml PDGF was obvious,and the OD450 value was higher,and the difference was statistically significant.Under the treatment of 20ng/ml PDGF,the proliferation of HPASMCs was promoted in a time gradient dependent manner.3.The effect of PDGF on the migration of HPASMCs was detected by scratch test.The results showed that compared with the control group,the migration ability of human pulmonary artery smooth muscle cells was significantly enhanced after treated with 20ng/ml PDGF for 12 hours.4.The protein expressions of MGP and BMP4 in PDGF induced human pulmonary artery smooth muscle cells were detected by Western Blot.Compared with the control group,the expression of BMP4 was significantly increased and the expression of MGP was significantly decreased after 24 h of 20ng/ml PDGF induction.Compared with the control group,the protein expression of BMP4 showed no statistically significant difference or decrease compared with the control group under the conditions of 20ng/ml PDGF induction for 48 h and 72 h,while the protein expression of MGP was decreased with statistically significant difference.Conclusions:1.PDGF can promote the proliferation and migration of HPASMCs in a time gradient dependent manner.2.In the PDGF-induced proliferation of HPASMCs,the expression of MGP was down-regulated and the expression of BMP4 was up-regulated,MGP may inhibit the PDGF-induced proliferation of HPASMCs by antagonizing BMP4.