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Aim To explore the effects of salvianolic acid B(SAB) and tanshinone ⅡA(TA) alone or the compatibility of these two effective components on the proliferation of rat vascular adventitial fibroblasts, and to observe the effects of the compatibility of the two on cell proliferation with the stimuation of angiotensin Ⅱ(Ang Ⅱ).Methods The effects of SAB and TA alone or the compatibility of the two on cell proliferation were detected by methyl thiazolyl tetrazolium(MTT) method, and flow cytometry was adopted to detect cell cycle with or without the induction of Ang Ⅱ.Results It was shown that SAB and TA alone could inhibit fibroblast proliferation in different degree.Through a series of concentration screening, three effective concentrations were obtained respectively and then the inhibition of cell growth was detected by Pairwise compatibilities.The results showed that the compatibility of TA(10-8 mol·L-1) and SAB(10-5 mol·L-1) had the most significant inhibitory effect(P<0.01), and they could inhibit cell proliferation, further flow cytometry was adopted to detect drug effects on cell cycle, the results indicated that the compatibility of SAB and TA mainly blocked the cells in G0/G1 phase.Induced by Ang Ⅱ, the compatibility of SAB and TA group, compared with Ang Ⅱ group, blocked thee cells in G0/G1 phase;and compared with combination of SAB and TA group, it mainly blocked cell cycle in S phase.Conclusion SAB and TA have certain inhibitory effect on fibroblast proliferation, also they could inhibit the proliferation induced by Ang Ⅱ, mainly by blocking the cells in G0/G1 phase.
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AIM:To explore the effect of angiotensin II ( Ang II) on phenotypic transformation in adventitial fibroblast subpopulations isolated from rat thoracic aorta .METHODS: Vascular adventitial fibroblasts were individually expanded by the method of cloning rings .The isolated cells were identified by reverse transcription-polymerase chain reac-tion ( RT-PCR) and immunofluorescence staining .The rat vascular adventitial fibroblasts were cultured in vitro and incuba-ted with or without Ang II.Using the methods of fluorescence quantitative polymerase chain reaction (FQ-PCR), immuno-fluorescence staining and Western blot , the expression of α-smooth muscle actin (α-SMA) was detected .RESULTS:The spindle cells and round cells were isolated by the method of cloning rings .From passage 3 to passage 8,α-SMA expression was decreased in the spindle cells , and increased significantly in the round cells .Induced by Ang II , the expression of α-SMA was increased in the 2 cell subpopulations, and that in the round cells was significantly increased (P<0.01).The results of Western blot indicated that Ang II stimulated the subpopulation of the round cells to transfer to myofibroblasts . CONCLUSION:Ang II promotes the phenotypic transformation of adventitial fibroblast subpopulations , and the 2 subpop-ulations may play an important role in vascular remodeling and reparative process .
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The study is to investigate the effect of angiotensin II (Ang II) and its receptor blockers on migration and endothelin-1 (ET-1) expression of rat vascular adventitial fibroblast subpopulations. Vascular adventitial fibroblasts were individually expanded by using cloning rings, and the effects of Ang II on the migration of adventitial fibroblast subpopulations were evaluated by Transwell. Fluorescence quantitative-PCR detected the expression of preproET-1 mRNA induced by Ang II, and its receptor antagonists losartan and PD-123319. The concentration of ET-1 was determined by ELISA. It showed that spindle shaped and epithelioid shaped cells were isolated by using cloning rings, named as spindle cells and round cells. RT-PCR showed that fibroblast subpopulations did not have leukocytes, endothelial cells and smooth muscle cells, namely pure cell lines. Compared with respective control cells, two subpopulations had transferring ability. Ang II significantly improved round cells migration in a concentration-dependent manner, and had no obvious influence on spindle cells migration. Ang II (1 x 10(-8) - 1 x 10(-6) mol x L(-1)) significantly increased the expression of preproET-1 mRNA in round cells (P < 0.01), and had no significant effect on the expression of preproET-1 mRNA in spindle cells. Losartan blocked the expression of preproET-1 mRNA induced by Ang II in round cells, and had no significant effect on the expression of preproET-1 mRNA in spindle cells. The effects of Ang II and ET-1 receptor inhibitors on the release of ET-1 were similar to the expression of preproET-1 mRNA. The results indicate that there are two cell subpopulations: round cells and spindle cells in rat vascular adventitial fibroblasts. Ang II significantly improved cells migration, and increased the expression of ET-1 in round cell subpopulation. It suggested that there may be different migratory mechanisms in two cell subpopulations, and the two subpopulations may play a different role in vascular remodeling and reparative process.
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Objective To investigate the relationship between polymorphism of the estrogen receptor(ER) gene and bone mineral density(BMD), and bone metabolism in Chinese postmenopausal healthy women. Methods In 246 postmenopausal healthy women, aged 44-78 years (average 61 years), BMD of lumbar spine and femoral neck, Ward and trochanteric areas were measured by dual energy X ray absorptiometry. Biochemical indexes(serum Ca, P, ALP, PTH, CT)were measured and PvuⅡ and XbaⅠ restriction fragment length polymorphism(RFLP) of ER were analysed by polymerase chain reaction restriction fragment length polymorphism(PCR RFLP). The relationship between polymorphism of the ER gene and BMD, and bone metabolism were evaluated. The RFLP was represented as Pp(PvuⅡ) and Xx(XbaI). Results In the PPxx genotype, Z score values of BMD were significantly lower than those in other genotypes. Conclusion RFLP of ER gene associated with BMD in postmenopausal healthy women and this might explain the cause of postmenopausal osteoporosis in Chinese women.
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Receptor for advanced glycation end products(RAGE) is a member of the immunoglobulin superfamily.In central nervous system,RAGE is mainly expressed by neurons,microglia,and cerebral endothelial cells.In Alzheimers disease(AD) brain,levels of RAGE are significantly increased from the positive feedback mechanisms driven by excess amounts of ?-amyloid protein(A?).The interaction of RAGE with A? in neurons,microglia,and cerebral endothelial cells induces the perturbation of neuronal functions,amplification of microglia inflammatory responses,and vascular dysfunction.Further understanding the molecular mechanisms associating RAGE-A? interaction provides us the opportunity to develop the therapeutic approaches for the devastating disease.