Effect of metformin on TGF-β1-induced fibrosis in human endometrial stromal cells / 解放军医学杂志

[Abstract] Objective To explore the effect of metformin on the fibrosis of human endometrial stromal cells (HESCs) induced by transforming growth factor-β1 (TGF-β1). Methods The in vitro model of endometrial fibrosis was established using TGF-β1 treated HESCs, and then treated with TGF-β1 in presence or absence of metformin at different concentrations (10, 50, 200 μmol/L). There set 5 experimental groups: normal control group, model group, 10 μmol/L metformin intervention group, 50 μmol/L metformin intervention group, and 200 μmol/L metformin intervention group. The mRNA and protein expression levels of α-smooth muscle actin (α-SMA), collagen type 1 alpha 1 (Col1α1) and fibronectin were detected by qPCR and Western blotting. The cell proliferation was detected by CCK-8 and EdU staining. Results The mRNA expression levels of α-SMA in the 5 groups were 1.003±0.108, 4.329±0.342, 3.383±0.342, 2.310±0.263 and 1.566±0.205; of Col1α1 were 1.002±0.299, 3.462±0.695, 2.342±0.531, 1.970±0.336 and 1.450±0.233; and of fibronectin were 1.000±0.225, 4.455±0.524, 3.164±0.317, 2.728±0.311 and 2.145±0.218; the differences between groups were statistically significant (P0.05). The OD values of the 5 groups were 0.313±0.091, 1.497±0.081, 1.095±0.059, 0.895±0.046 and 0.560±0.082, and the positive rate of EdU cells were 12.665±3.146, 47.650±4.656, 37.568±2.549, 32.050±3.652 and 26.050±3.453, showing statistically significant differences between the groups (P<0.001). Compared with normal control group, the proliferation capacity of model group increased significantly (P<0.01). Compared with model group, the proliferation capacity of 10, 50 and 200 μmol/L metformin intervention group reduced significantly in a dose-dependent manner (P<0.01). Conclusion Metformin may play an anti-fibrosis effect via inhibiting TGF-β1-induced myofibroblast differentiation as well as ECM synthesis and proliferation in HESCs.

Elementos da matriz extracelular da cérvice uterina e trabalho de parto prematuro / Elements of extracellular matrix from uterine cervix and premature labor

As taxas de prematuridade não atingem quedas significativas, mesmo em países desenvolvidos, alertando os pesquisadores a buscarem alternativas clínico-farmacológicas na prevenção desta enfermidade obstétrica. A matriz extracelular ocupa espaço relevante na maturação cervical, e seu entendimento é fundamental para possíveis ações preventivas. Destaca-se a provável correlação dos níveis locais de glicosaminoglicanos na prevenção do parto prematuro por inibir processos inflamatórios na cérvice uterina

The preterm delivery rates do not decrease significantly, even in developed countries, prompting researchers to seek pharmacological and clinical alternatives to prevent this obstetrics disorder. The extracellular matrix is important in cervical length, and its understanding is essential for possible preventive actions. We emphasize the probable correlation of the local levels of glycosaminoglycans to prevent premature labor by inhibiting inflammatory processes from uterine cervix

NG2 proteoglycan promotes mesangial cells proliferation and extracellular matrix production / 中华肾脏病杂志

Objective To explore the role of NG2 proteoglycan in the pathogenesis of glomerulosclerosis. Methods Eukaryotic expression vectors carrying the small hairpin RNA (shRNA) for NG2 mRNA , named as Psilencer-NG2, was constructed. Then, rat mesangial cells (RMC) were transfeeted with Psilencer-NG2, Psilencer-NC (negative control), pcDNA/NG2 (NG2 over-expressive vector) and empty vector pcDNA 1 respectively. The expression of endogenous NG2 in RMCs was examined by real-time PCR and Western blotting. Cell proliferation was analyzed by MTT assay and flow cytometry. The expression of laminin was detected by real-time PCR. Results Transfection of pcDNA/NG2 into HBZY-1 cells resulted in over-expression of NG2 mRNA and protein (P<0.05, P<0.05). Transfection of Psilencer-NG2 led to reduced expression of NG2 mRNA and protein (P<0.01, P<0.01). The expression of laminin β1 significantly increased due to overexpression of NG2 and decreased by treating with NG2 siRNA. According to MTT assay, overexpreasion of NG2 significantly stimulated the proliferation of mesangial cells while NG2 silencing inhibited it. NG2 increased the cell number in S phase and decreased the cell number in G0/G1 phase, while silencing NG2 induced the decrease of cell number in S phase and the increase of cell number in G0/G1 phase. Conclusion NG2 actively participates in the pathogenesis of glomerulosclerosis by stimulating proliferation of RMCs and increasing the deposition of ECM.

A novel scaffold for endothelial progenitor cells in tissue engineered vascular grafts / 中华普通外科杂志

Objective To explore the characteristics of endothelial progenitor cells (EPCs) on different scaffolds and to find a new bio-engineered synthetic hybrid scaffold for artificial bio-engineered blood vessels. Methods EPCs were induced from mesenchymal stem cells isolated from rat bone marrow and seeded on ECM scaffold. The surface structure of the scaffold and growth status of EPCs on the scaffold were observed and analysed by electron microscopy. The characteristics and number of those EPCs on different kinds of scaffolds were studied with EPC-specific VWF by immunofluorescence, Western blotting and real-time PCR technique at different time points. Results The cell adhesion rate at 1,3,5 h after seeded on pressed scaffold were higher than that on unpressed scaffolds( P < 0. 01 ). Pressed scaffolds has got a larger cell number( P < 0. 05 )at DIV1, DIV3, DIV7, but there was no significant difference after DIV10. Furthermore, cell shapes of EPCs on pressed scaffolds were more mature and more similar to endothelial cells. A level cell surface on pressed scaffolds was achieved. Western blotting assays revealed EPCs on pressed scaffolds expressed more protein VWF at DIV3, DIV7, DIV10. Real-time PCR results showed EPCs on the two different groups of scaffolds all expressed VWF gene, The quantity of their expression in the two groups were all enhanced after DIV7 (P < 0. 05 ). The quantity of VWF gene expression in the pressed group was much higher than that in the unpressed group at DIV3 ,DIV7,DIV10 (P <0, 01), but there was no significant difference after DIV14. Conclusions Pressed ECM scaffolds can promote adhesion, proliferation and differentiation on EPCs. Pressed scaffolds can be used as the matrix for EPC and fabricated into a novel synthetic tissue bio-engineered vascular scaffold.

Effects of Puerarin on Proliferation of Human Embro Fibroblast Proliferation and Extracelluar Matrix in Vitro / 中国中医药信息杂志

Objective To investigate the effect of puerarin on proliferation of human embro fibroblast and extracelluar matrix. Method Human embro fibroblasts were incubated with 0～400 ?g/mL puerarin for 24～96 h. The MTT method was used to assay the biological activities and extracelluar matrix formation in different time and different dose of pueratin. The cell cycle distribution was detected by flow cytometric analysis. Result Incubation cells in presence of puerarin of 40～400 ?g/mL for 24～72 h could significant inhibit cell proliferation and restrain the collagen synthesis in a dose- and time-dependent manner (P

The effect of hyaluronic acid on expression of extracellular matrix proteins and bone formation in rabbit mandibular distraction osteogenesis

Distraction osteogenesis is a new bone formation technique. There is a advantage of the environmental adaptation when distraction force is applied to the gap between osteotomy lines. But it has a disadvantage of long-term wearing of the appliance and long consolidation period. Therefore we make an effort to reduce it and repair normal function. Extracellular matrix proteins have a function to control the cellular growth, migration, shape and metabolism. In these, hyaluronic acid is a member of polysaccharide glycosaminoglycans (GAGs) and has a important function as bone formation and osteoinduction property. PURPOSE: In this experimental study in rabbit mandibular distraction osteogenesis, we investigated the bone enhancing property of hyaluronic acid and the expression of extracellular proteins such as osteocalcin and osteonectin. MATERIALS AND METHODS: The experimental study was carried out on 24 Korean male white rabbits (both mandibular body, n=48). Distraction group was divided to distraction experimental (A, n=16) and distraction control (B, n=16) by the application of hyaluronic acid (Hyruan, LGCI, Seoul, Korea). Normal control group (C, n=16) was only osteotomized. After 5 days latency, distraction devices were activated at a rate of 1.4 mm per day (0.7 mm every 12hours) for 3.5 days. Animals were sacrificed at postoperative 3, 7, 14, and 28 days. HandE stain and immunohistochemical stain was done on decalcified section. Additionally RT-PCR analysis was done for the identification of the expression of osteocalcin and osteonectin. RESULTS: The bone formation in distraction experimental group was much more than that in distraction and normal control group at postoperative 28 days. In immunohistochemical stain, osteocalcin was enhanced at only postoperative 14 days, but osteonectin was not different at each post-operation days. In RT-PCR analysis, osteocalcin was not different at each post-operation days, but osteonectin was strongly expressed in distraction experimental group at postoperative 7 days. The expression of osteocalcin and osteonectin was elevated during the healing period. CONCLUSION: We found the good bone formation ability of hyaluronic acid in distraction osteogenesis through the immunohistochemistry and RTPCR analysis to osteocalcin and osteonectin, known as a bone formation marker. The application of hyaluronic acid in distraction osteogenesis is a method to reduce the consolidation period.