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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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Aim To observe the effects of effective fraction of Epimedium,Astragalus,Radix Puerariae on behavioral and pathological changes in a transgenic mouse model of Alzheimer’s disease.Methods Six-month-old APPswe /PS1 ΔE9 transgenic mice were ran-domly divided into 2 groups:model group and effective fraction group,1 0 mice each group.The mice in the effective fraction group were treated with the effective fraction of Astragalus,Radix Puerariae,Epimedium compound for 8 weeks.The C57BL/6J mice were used as negative control group.After 8 weeks,the learning and memory function were measured by Morris water maze,the pathological changes in brain tissue were ob-served by Modified Bielschowsky staining and Nissl 's staining.Results During place navigation trial,the escape latency in the APPswe /PS1 ΔE9 double transgenic model mice was longer than those of the mice of C57 (P 0.05 ). The Modified Bielschowsky staining shows that the neuron fibers of the cerebral cortex of APPswe /PS1 ΔE9 double transgenic mice were enlarged,swelling,and dense.There were senile plaques and nerve fiber tangles in the cerebral cortex of APPswe /PS1 ΔE9 double transgenic mice.The neuron fibers of mice in the effective fraction group were relieved;there was a small amount of senile plaque.The Nissl’s staining shows that the neurons of the cerebral cortex of APPswe /PS1 ΔE9 mice were edema, the number of cells were decreased.The mice in the effective fraction group were free of the disease.Con-clusion The double transgenic APPswe /PS1 ΔE9 mice of AD can simulate the specific pathogenesis of AD, which may be the efficient experimental animal model. The effective fraction of epimedium,astragalus and ra-dix puerariae may have a neuroprotective effect against AD via improving the learning and memory ability,and reduce the cerebral cortex nerve fiber tangles,senile plaques and neurons edema changes.
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<p><b>OBJECTIVE</b>To study the effect and possible impact mechanism of salidroside on cognitive ability of Alzheimer's disease (AD) model rats induced by amyloid beta peptide (Abeta1-40).</p><p><b>METHOD</b>Abeta1-40 was injected into bilateral hippocampus to create the AD model. Afterwards, different doses of salidroside (25, 50, 75 mg x kg(-1)) were orally administered for 21 days. Rats' learning and memory abilities were detected by Morris water maze testing system. The levels of the superoxide dismutase (SOD), malondialdehyde (MDA), and the expression of nuclear factor-kappaB (NF-kappaB), inducible nitric oxide synthase (iNOS) and receptor for advanced glycation end products (RAGE) protein in hippocampus were also detected by different methods.</p><p><b>RESULT</b>The place navigation test showed longer escape latency, low frequency of platform quadrant crossing per unit time, damage in learning capacity, significant decrease in SOD acivity in hippocampus, notable increase in MDA content, NF-kappaB, iNOS and RAGE protein expressions in rats. Salidroside (50, 75 mg x kg(-1)) significantly alleviated the impairments of learning and memory ability. The activity of SOD increased in salidroside (50 droside group compared with that of the Alzheimer's disease group (P < 0.01).</p><p><b>CONCLUSION</b>Salidroside may treat Alzheimer's disease by inhibiting the oxidative stress.</p>
Subject(s)
Animals , Male , Rats , Alzheimer Disease , Drug Therapy , Amyloid beta-Peptides , Toxicity , Cognition , Disease Models, Animal , Glucosides , Pharmacology , Therapeutic Uses , Maze Learning , NF-kappa B , Metabolism , Nitric Oxide , Physiology , Phenols , Pharmacology , Therapeutic Uses , Receptor for Advanced Glycation End Products , Receptors, Immunologic , Superoxide Dismutase , MetabolismABSTRACT
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.