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Objective To investigate the effect and mechanism of Yes-associated protein (YAP) on rat cardiac fibroblasts(CFs) proliferation and transdifferentiation induced by high glucose. Methods The CFs cells were isolated and cultured of newborn SD rats aged 1-3 days, then treated with 40 mmol/L D-glucose for concentration of diabetes cardiomyopathy model(DCM), and the relative expression levels were detected with Western blotting of YAP, α-smooth muscle actin (α-SMA), collagenⅠ, collagen Ⅲ and connective tissue growth factor (CTGF), respectively, at the time points 24 hours. The CFs cells were divided into normal glucose (NG) control group (5.5 mmol/L D-glucose), high glucose (HG) group (40.0 mmol/L D-glucose), normal glucose +0.5 μmol/L verteporfin (NG+VP) group and high glucose + 0.5 μmol/L verteporfin (HG+VP) group. Verteporfin is an inhibitor of YAP, which blockades the interaction between YAP and TEAD. Immunofluorescence of vimentin was used to identify CFs. Immunofluorescence of Ki-67 was used to detect the proliferation activities of CFs in various groups. Western blotting was performed to detect the levels of α-SMA, collagen Ⅰ and collagen Ⅲ, YAP and CTGF proteins in CFs of various groups. Results The positive result of vimentin immunofluorescence prompted that the primary cultured cells were rat's CFs. Immunofluorescence of Ki-67 showed that, the positive rate (%) of Ki-67 in CFs was obviously higher in HG group than in NG group (67.33±5.14 vs. 22.94±4.88, P<0.05); After treatment with VP, the positive rate (%) of Ki-67 in CFs decreased markedly (46.83±3.86 vs. 67.33±5.14, P<0.05). Western blotting showed that, compared with NG group, the relative expression levels of α-SMA, collagenⅠ and collagen Ⅲ in CFs increased significantly in HG group (1.43±0.98 vs. 0.93±0.06, 1.80±0.09 vs. 1.08±0.09, 1.43±0.09 vs. 0.88±0.10, P<0.05), and the relative expression levels of VAP and CTGF protein also increased (1.93±0.15 vs. 1.17±0.09, 1.80±0.18 vs. 1.23±0.16, P<0.05). Compared with the HG group, the relative expression levels of α-SMA, collagen I, collagen Ⅲand CTGF proteins decreased significantly in HG+VP group (1.27±0.06 vs. 1.71±0.12, 2.05±0.23 vs. 3.03±0.17, 1.10±0.12 vs. 1.82±0.18, 1.31±0.16 vs. 1.57±0.03, P<0.05), while compared with NG group, the relative expression levels of α-SMA, collagen Ⅰ and CTGF proteins were higher in HG+VP group (P<0.05). Conclusion High glucose promotes the proliferation and activation of neonatal rats CFs and excessive synthesis of extracellular matrix proteins such as collagen by regulating the YAP/TEAD/CTGF signaling pathway.
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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