白杨素对动脉损伤后新生内膜增生的影响及机制
|
摘要
目的利用小鼠颈动脉导丝损伤的动物模型和血小板衍生生长因子(PDGF)-BB刺激血管平滑肌细胞(VSMCs)增殖的细胞模型,研究白杨素对动脉损伤后新生内膜增生的抑制作用及相关机制。方法建立小鼠颈动脉导丝损伤模型,随机分成模型对照组与白杨素组,分别喂以正常啮齿类动物饲料和含0. 09%白杨素[w/w,相当于150mg/(kg·d)白杨素]的正常啮齿类动物饲料。取术后28天损伤侧颈动脉,检测指标。苏木素-伊红染色评价新生内膜增生,测算内膜面积和内膜中膜比值;免疫组化法测定损伤动脉内增殖细胞核抗原(PCNA)阳性细胞表达状态,评价白杨素对内膜增生的影响。同时体外培养VSMCs,给予20ng/ml PDGF-BB和(或) 12. 5μmol/L白杨素处理,流式细胞仪检测细胞增殖,Western blot法测定c-Jun氨基末端激酶(JNK)和信号转导及转录激活因子(STAT) 3的总蛋白和磷酸化蛋白水平。结果白杨素显著抑制导丝损伤所致颈动脉内膜增生,降低内膜面积、内中膜比值以及损伤侧颈动脉新生内膜内PCNA阳性细胞表达数。PDGF-BB明显促进VSMCs增殖,并显著升高VSMCs中JNK和STAT3的磷酸化水平,白杨素导致PDGF-BB引起的VSMCs增殖停滞在G0/G1期,并显著减弱PDGF-BB诱导的JNK和STAT3磷酸化。结论白杨素可能通过阻止JNK和STAT3通路活化降低VSMCs增殖,从而遏制动脉损伤后新生内膜增生。
Objective To investigate the effects and possible mechanism of chrysin on neointimal hyperplasia after artery injury through guide wire-injured murine carotid artery in vivo and platelet derived growth factor( PDGF-BB)-treated vascular smooth muscle cells( VSMCs) in vitro. Methods Male C57 BL/6 mice aged 10 weeks underwent operation of guide wire injury in left carotid arteries,and then the mice were randomly divided into injured control group and chrysin group. After the surgery,the mice were fed either normal rodent chow or normal chow containing 0. 09% chrysin [w/w,which corresponded to 150 mg/( kg·d) chrysin if a 30 g mouse consumed 5 g of chow]for 28 d before they were euthanized. The left carotid arteries were isolated and processed for pathological analysis. Using hematoxylin-eosin( HE) staining,the intimal( I) and medial( M) areas were measured and I/M ratios were calculated. Proliferating cell nuclear antigen( PCNA) immunohistochemistry staining was performed to evaluate neointimal cell proliferation. Furthermore,cultured VSMCs were treated with 20 ng/ml PDGF-BB and/or 12. 5μmol/L chrysin pretreatment,flow cytometric analysis was carried out to detect cell proliferation,Western blot was performed to evaluate total and phosphorylated protein levels of c-Jun amino-terminal kinase( JNK) and signal transducers and activators of transcription 3( STAT3). Results Morphologic analysis based on HE staining of carotid cross-sections showed that the neointimal area and I/M ratio in the chrysin group were significantly reduced compared with the injured controls at day 28 after carotid injury. Chrysin also markedly suppressed neointimal cell proliferation,as evidenced by reduced numbers of PCNA-positive cells in the neointima. 20 ng/ml PDGF-BB induced VSMCs proliferation and a significant increase in the levels of phosphorylated JNK and STAT3. 12. 5μmol/L chrysin pretreatment caused PDGF-BB-stimulated VSMCs to arrest at G0/G1 phases and attenuated the PDGF-BB-induced JNK and STAT3 activation. Conclusion Chrysin may suppress VSMCs proliferation via the inhibition of the JNK and STAT3 signaling pathways,and therefore inhibit neointima hyperplasia after artery injury.
Objective To investigate the effects and possible mechanism of chrysin on neointimal hyperplasia after artery injury through guide wire-injured murine carotid artery in vivo and platelet derived growth factor( PDGF-BB)-treated vascular smooth muscle cells( VSMCs) in vitro. Methods Male C57 BL/6 mice aged 10 weeks underwent operation of guide wire injury in left carotid arteries,and then the mice were randomly divided into injured control group and chrysin group. After the surgery,the mice were fed either normal rodent chow or normal chow containing 0. 09% chrysin [w/w,which corresponded to 150 mg/( kg·d) chrysin if a 30 g mouse consumed 5 g of chow]for 28 d before they were euthanized. The left carotid arteries were isolated and processed for pathological analysis. Using hematoxylin-eosin( HE) staining,the intimal( I) and medial( M) areas were measured and I/M ratios were calculated. Proliferating cell nuclear antigen( PCNA) immunohistochemistry staining was performed to evaluate neointimal cell proliferation. Furthermore,cultured VSMCs were treated with 20 ng/ml PDGF-BB and/or 12. 5μmol/L chrysin pretreatment,flow cytometric analysis was carried out to detect cell proliferation,Western blot was performed to evaluate total and phosphorylated protein levels of c-Jun amino-terminal kinase( JNK) and signal transducers and activators of transcription 3( STAT3). Results Morphologic analysis based on HE staining of carotid cross-sections showed that the neointimal area and I/M ratio in the chrysin group were significantly reduced compared with the injured controls at day 28 after carotid injury. Chrysin also markedly suppressed neointimal cell proliferation,as evidenced by reduced numbers of PCNA-positive cells in the neointima. 20 ng/ml PDGF-BB induced VSMCs proliferation and a significant increase in the levels of phosphorylated JNK and STAT3. 12. 5μmol/L chrysin pretreatment caused PDGF-BB-stimulated VSMCs to arrest at G0/G1 phases and attenuated the PDGF-BB-induced JNK and STAT3 activation. Conclusion Chrysin may suppress VSMCs proliferation via the inhibition of the JNK and STAT3 signaling pathways,and therefore inhibit neointima hyperplasia after artery injury.
引文
1 Shi N,Chen SY. Mechanisms simultaneously regulate smooth muscle proliferation and differentiation[J]. J Biomed Res,2014,28(1):40-46
2 Li PC,Sheu MJ,Ma WF,et al. Anti-restenotic roles of dihydroaustrasulfone alcohol involved in inhibiting PDGF-BB-stimulated proliferation and migration of vascular smooth muscle cells[J]. Mar Drugs,2015,13(5):3046-3060
3 Kim JY,Kim KH,Lee WR,et al. Apamin inhibits PDGF-BB-induced vascular smooth muscle cell proliferation and migration through suppressions of activated Akt and Erk signaling pathway[J]. Vascul Pharmacol,2015,70:8-14
4 Neeli I,Liu Z,Dronadula N et al. An essential role of the Jak-2/STAT-3/cytosolic phospholipase A(2)axis in platelet-derived growth factor BB-induced vascular smooth muscle cell motility[J].J Biol Chem,2004,279(44):46122-46128
5 Rehman MU,Ali N,Rashid S,et al. Alleviation of hepatic injury by chrysin in cisplatin administered rats:probable role of oxidative and inflammatory markers[J]. Pharmacol Rep,2014,66(6):1050-1059
6 Gresa-Arribas N,Serratosa J,Saura J,et al. Inhibition of CCAAT/enhancer binding protein delta expression by chrysin in microglial cells results in anti-inflammatory and neuroprotective effects[J]. J Neurochem,2010,115(2):526-536
7 Weng MS,Ho YS,Lin JK. Chrysin induces G1 phase cell cycle arrest in C6 glioma cells through inducing p21Waf1/Cip1 expression:involvement of p38 mitogen-activated protein kinase[J]. Biochem Pharmacol,2005,69(12):1815-1827
8 Kasala ER,Bodduluru LN,Madana RM,et al. Chemopreventive and therapeutic potential of chrysin in cancer:mechanistic perspectives[J]. Toxicol Lett,2015,233(2):214-225
9 秦小江,侯晓敏,梁泰刚.白杨素对大鼠离体肾动脉的舒张作用[J].中国药物与临床,2014,14(8):1009-1011
10 Pushpavalli G,Veeramani C,Pugalendi KV. Influence of chrysin on hepatic marker enzymes and lipid profile against D-galactosamineinduced hepatotoxicity rats[J]. Food Chem Toxicol,2010,48(6):1654-1659
11 Zou XQ,Peng SM,Hu CP,et al. Synthesis,characterization and vasculoprotective effects of nitric oxide-donating derivatives of chrysin[J]. Bioorg Med Chem,2010,18(9):3020-3025
12 El-Bassossy HM,Abo-Warda SM,Fahmy A. Chrysin and luteolin alleviate vascular complications associated with insulin resistance mainly through PPAR-γactivation[J]. Am J Chin Med,2014,42(5):1153-1167
13 严玲,廖正凯,关红菁,等.白杨素抑制血小板源性生长因子诱导的血管平滑肌细胞迁移和表型转换[J].医学研究杂志,2016,45(1):25-28
14 严玲,廖正凯,关红菁,等.白杨素抑制血小板源性生长因子诱导的血管平滑肌细胞增殖的机制研究[J].医学研究杂志,2015,44(12):34-37
15 Guan H,Chen C,Zhu L,et al. Indole-3-carbinol blocks plateletderived growth factor-stimulated vascular smooth muscle cell function and reduces neointima formation in vivo[J]. J Nutr Biochem,2013,24(1):62-69
16 Guan H,Gao L,Zhu L,et al. Apigenin attenuates neointima formation via suppression of vascular smooth muscle cell phenotypic transformation[J]. J Cell Biochem,2012,113(4):1198-1207
17 朱丽华,王朗,陈静,等.小鼠颈动脉导丝损伤模型建立与特点分析研究[J].岭南心血管病杂志,2012,18(4):431-437
18 Lee SJ,Yoon JH,Song KS. Chrysin inhibited stem cell factor(SCF)/c-Kit complex-induced cell proliferation in human myeloid leukemia cells[J]. Biochem Pharmacol,2007,74(2):215-225
19 Lin CM,Shyu KG,Wang BW,et al. Chrysin suppresses IL-6-induced angiogenesis via down-regulation of JAK1/STAT3 and VEGF:an in vitro and in ovo approach[J]. J Agric Food Chem,2010,58(11):7082-7087
2 Li PC,Sheu MJ,Ma WF,et al. Anti-restenotic roles of dihydroaustrasulfone alcohol involved in inhibiting PDGF-BB-stimulated proliferation and migration of vascular smooth muscle cells[J]. Mar Drugs,2015,13(5):3046-3060
3 Kim JY,Kim KH,Lee WR,et al. Apamin inhibits PDGF-BB-induced vascular smooth muscle cell proliferation and migration through suppressions of activated Akt and Erk signaling pathway[J]. Vascul Pharmacol,2015,70:8-14
4 Neeli I,Liu Z,Dronadula N et al. An essential role of the Jak-2/STAT-3/cytosolic phospholipase A(2)axis in platelet-derived growth factor BB-induced vascular smooth muscle cell motility[J].J Biol Chem,2004,279(44):46122-46128
5 Rehman MU,Ali N,Rashid S,et al. Alleviation of hepatic injury by chrysin in cisplatin administered rats:probable role of oxidative and inflammatory markers[J]. Pharmacol Rep,2014,66(6):1050-1059
6 Gresa-Arribas N,Serratosa J,Saura J,et al. Inhibition of CCAAT/enhancer binding protein delta expression by chrysin in microglial cells results in anti-inflammatory and neuroprotective effects[J]. J Neurochem,2010,115(2):526-536
7 Weng MS,Ho YS,Lin JK. Chrysin induces G1 phase cell cycle arrest in C6 glioma cells through inducing p21Waf1/Cip1 expression:involvement of p38 mitogen-activated protein kinase[J]. Biochem Pharmacol,2005,69(12):1815-1827
8 Kasala ER,Bodduluru LN,Madana RM,et al. Chemopreventive and therapeutic potential of chrysin in cancer:mechanistic perspectives[J]. Toxicol Lett,2015,233(2):214-225
9 秦小江,侯晓敏,梁泰刚.白杨素对大鼠离体肾动脉的舒张作用[J].中国药物与临床,2014,14(8):1009-1011
10 Pushpavalli G,Veeramani C,Pugalendi KV. Influence of chrysin on hepatic marker enzymes and lipid profile against D-galactosamineinduced hepatotoxicity rats[J]. Food Chem Toxicol,2010,48(6):1654-1659
11 Zou XQ,Peng SM,Hu CP,et al. Synthesis,characterization and vasculoprotective effects of nitric oxide-donating derivatives of chrysin[J]. Bioorg Med Chem,2010,18(9):3020-3025
12 El-Bassossy HM,Abo-Warda SM,Fahmy A. Chrysin and luteolin alleviate vascular complications associated with insulin resistance mainly through PPAR-γactivation[J]. Am J Chin Med,2014,42(5):1153-1167
13 严玲,廖正凯,关红菁,等.白杨素抑制血小板源性生长因子诱导的血管平滑肌细胞迁移和表型转换[J].医学研究杂志,2016,45(1):25-28
14 严玲,廖正凯,关红菁,等.白杨素抑制血小板源性生长因子诱导的血管平滑肌细胞增殖的机制研究[J].医学研究杂志,2015,44(12):34-37
15 Guan H,Chen C,Zhu L,et al. Indole-3-carbinol blocks plateletderived growth factor-stimulated vascular smooth muscle cell function and reduces neointima formation in vivo[J]. J Nutr Biochem,2013,24(1):62-69
16 Guan H,Gao L,Zhu L,et al. Apigenin attenuates neointima formation via suppression of vascular smooth muscle cell phenotypic transformation[J]. J Cell Biochem,2012,113(4):1198-1207
17 朱丽华,王朗,陈静,等.小鼠颈动脉导丝损伤模型建立与特点分析研究[J].岭南心血管病杂志,2012,18(4):431-437
18 Lee SJ,Yoon JH,Song KS. Chrysin inhibited stem cell factor(SCF)/c-Kit complex-induced cell proliferation in human myeloid leukemia cells[J]. Biochem Pharmacol,2007,74(2):215-225
19 Lin CM,Shyu KG,Wang BW,et al. Chrysin suppresses IL-6-induced angiogenesis via down-regulation of JAK1/STAT3 and VEGF:an in vitro and in ovo approach[J]. J Agric Food Chem,2010,58(11):7082-7087
© 2004-2018 中国地质图书馆版权所有 京ICP备05064691号 京公网安备11010802017129号 地址：北京市海淀区学院路29号 邮编：100083 电话：办公室：(+86 10)66554848；文献借阅、咨询服务、科技查新：66554700 |