Pirfenidone suppresses the abnormal activation of human Müller cells after platelet-derived growth factor-BB stimulation

AIM: To determine the effect of pirfenidone on the activated human Müller cells by platelet-derived growth factor-BB(PDGF-BB). METHODS: The primary human Müller cells were separated from retinal tissues and established the pathogenic model by stimulated with PDGF-BB. The Müller cells behaviour of normal group and the model group was measured by MTT assay, Trypan blue assay, cell migration assay, and collagen contraction assay. The expression of transforming growth factor(TGF)-β1,-β2, and pigment epithelium-derived factor(PEDF) was estimated with realtime polymerase chain reaction(PCR), Western blot and immunofluorescence analyses. RESULTS: A pathogenic/proliferative model of Müller cells was established by stimulating normal cultured Müller cells with 10 ng/mL PDGF-BB for 48 h. After treated with 0.2 and 0.3 mg/mL pirfenidone, the proliferation, migration and collagen contraction was statistically significantly depressed in the model group compared with the normal groups. The expression levels of TGF-β1 and TGF-β2 were significantly down-regulated, while the PEDF expression was significantly up-regulated after treated with 0.2 and 0.3 mg/mL pirfenidone in the model group. CONCLUSION: Pirfenidone effectively suppress the proliferation, migration and collagen contraction of the human Müller cells stimulated with PDGF-BB through down-regulation of TGF-β1/TGF-β2 and up-regulation of PEDF.

Effect of Korean Magnolia obovata Extract on Platelet-Derived Growth Factor-Induced Vascular Smooth Muscle Cells

Objective:To investigate the effects of Korean Magnolia obovata crude extract(KME)on plateletderived growth factor(PDGF)-BB-induced proliferation and migration of vascular smooth muscle cells(VSMCs).Methods:KME composition was analyzed by high-performance liquid chromatography(HPLC).VSMCs were isolated from the aorta of a Sprague-Dawley rat,incubated in serum free-Dulbecco's modified Eagle's medium in the presence or absence of KME(10,30,100,and 300(xg/mL),then further treated with PDGF-BB(10 ng/mL).VSMC proliferation was detected using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and VSMC migration was determined using the Boyden chamber and scratch wound healing assays.Western blot analysis was used to detect phosphorylation of extracellular signal-regulated protein kinases 1 and 2(p-ERK1/2),protein kinase B(p-Akt),and stress-activated protein kinase/c-Jun NH2-terminal kinase(p-SAPK/JNK).The antimigration and proliferation effects of KME were tested using aortic sprout outgrowth.Results:The HPLC analysis identified honokiol(0.45 mg/g)and magnolol(0.34 mg/g)as the major components of KME.KME(30,100,and 300μg/m L)significantly decreased the proliferation and migration of PDGF-BB-stimulated(10 ng/mL)VSMCs and the PDGF-BB-induced phosphorylation of EKR1/2,Akt,and SAPK/JNK(P<0.05).Furthermore,PDGF-BBinduced VSMCs treated with 300μg/m L of KME showed reduction in aortic sprout outgrowth.Conclusion:KME could inhibit abnormal proliferation and migration of VSMCs by down-regulating the phosphorylation of EKR1/2 and Akt.Thus,KME might be a functional food for preventing vascular disorders.