右归丸药物血清对高糖缺氧条件下大鼠视网膜微血管内皮细胞VEGF、VEGFR-2、PI3K、Akt基因及蛋白表达的影响
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摘要
目的:观察右归丸药物血清对高糖缺氧条件下SD大鼠视网膜微血管内皮细胞血管内皮生长因子(VEGF)、血管内皮生长因子受体2(VEGFR-2)、磷脂酰肌醇3激酶(PI3K)、蛋白激酶B(Akt)基因及蛋白表达的影响。方法:采用33mmol/L的葡萄糖和100μmol/L的CoCl2同时附加于培养基中培养细胞4d,以建立体外高糖缺氧的细胞模型,应用RT-PCR法及Western blot法检测各组右归丸药物血清对模型细胞VEGF、VEGFR-2、PI3K、Akt基因及蛋白表达水平的影响。结果:高糖及高糖缺氧均能导致大鼠视网膜微血管内皮细胞中VEGF、VEGFR-2、PI3K、Akt mRNA的表达量上升(P<0.003),而低、中、高剂量组的右归丸药物血清均可降低此高表达(P<0.003)。高糖缺氧亦可引起模型细胞中VEGF、VEGFR-2、PI3K及Ak蛋白表达增多(P<0.003),而中、高剂量组的右归丸药物血清均可降低此高表达(P<0.003)。结论:右归丸药物血清可能通过降低高糖缺氧条件下SD大鼠视网膜微血管内皮细胞中VEGF、VEGFR-2、PI3K、Akt基因及蛋白表达水平,从而发挥其抑制视网膜微血管内皮细胞增殖及迁移的作用,抑制新生血管生成。
Obiective: To observe the effects of Yougui Pills medical serum on mRNA and protein expression of VEGF,VEGFR-2, PI3 K and Akt in rat retinal microvascular endothelial cell under high glucose hypoxia condition. Methods: The cell high glucose and hypoxia model in vitro was established with 33 mmol/L glucose and 100μmol/L CoCl2 simultaneously cultured for4 days. Gene and protein expression levels of VEGF, VEGFR-2, PI3 K, Akt in model cells were detected by RT-PCR method and Western blot method. Results: High glucose and high glucose hypoxia could increase the mRNA expression of VEGF, VEGFR-2,PI3 K and Akt mRNA in rat retinal microvascular endothelial cell(P<0.003). While the low, middle and high dose medical serum of Yougui Pills could reduce those high expression(P<0.003). High glucose and hypoxia could also increase the protein expression of VEGF, VEGFR-2, PI3 K and Akt in model cells(P<0.003), in the middle and high dose groups, the medicinal serum of Yougui Pills could reduce this high expression(P<0.003). Conclusion: The medicinal serum of Yougui Pills played a role in inhibiting the proliferation and migration of retinal microvascular endothelial cells and inhibits neovascularization by reducing the mRNA and protein expression of VEGF, VEGFR-2, PI3 K and Akt and the expres in SD rat retinal microvascular endothelial cells under high glucose and hypoxia condition.
Obiective: To observe the effects of Yougui Pills medical serum on mRNA and protein expression of VEGF,VEGFR-2, PI3 K and Akt in rat retinal microvascular endothelial cell under high glucose hypoxia condition. Methods: The cell high glucose and hypoxia model in vitro was established with 33 mmol/L glucose and 100μmol/L CoCl2 simultaneously cultured for4 days. Gene and protein expression levels of VEGF, VEGFR-2, PI3 K, Akt in model cells were detected by RT-PCR method and Western blot method. Results: High glucose and high glucose hypoxia could increase the mRNA expression of VEGF, VEGFR-2,PI3 K and Akt mRNA in rat retinal microvascular endothelial cell(P<0.003). While the low, middle and high dose medical serum of Yougui Pills could reduce those high expression(P<0.003). High glucose and hypoxia could also increase the protein expression of VEGF, VEGFR-2, PI3 K and Akt in model cells(P<0.003), in the middle and high dose groups, the medicinal serum of Yougui Pills could reduce this high expression(P<0.003). Conclusion: The medicinal serum of Yougui Pills played a role in inhibiting the proliferation and migration of retinal microvascular endothelial cells and inhibits neovascularization by reducing the mRNA and protein expression of VEGF, VEGFR-2, PI3 K and Akt and the expres in SD rat retinal microvascular endothelial cells under high glucose and hypoxia condition.
引文
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[16]罗向霞,段俊国,李响.糖尿病视网膜病变阳虚病机的代谢组学物质基础.中国中医眼科杂志,2013,23(12):394
[2]Mysona B A,Matragoon S,Stephens M,et al.Imbalance of the nerve growth factor and its precursor as a potential biomarker for diabetic retinopathy.Bio Med Research International,2015,2015:12.571456
[3]Zhou Y,Zhang Y,Shi K,et al.Body mass index and risk of diabetic retinopathy:A meta-analysis and systematic review.Medicine,2017,96(22):e6754
[4]Vasant More S,Kim I S,Choi D K.Recent update on the role of Chinese material medica and formulations in diabetic retinopathy.Molecules,2017,22(1):76
[5]Lechner J,O'Leary O E,Stitt A W.The pathology associated with diabetic retinopathy.Vision Research,2017,139:7-14
[6]Roy S,Kern T S,Song B,et al.Mechanistic insights into pathological changes in the diabetic retina:Implications for targeting diabetic retinopathy.The American Journal of Pathology,2017,187(1):9-19
[7]孙晶丽.糖尿病视网膜病变的激光治疗临床疗效观察.中国现代药物应用,2016,10(16):113-114
[8]李欢.基于“阴中求阳”立法之右归丸对糖尿病视网膜病变大鼠VEGF-VEGFR促存活信号通路的调控作用研究.兰州:甘肃中医药大学,2016
[9]谢明捷.VEGF、IGF-1在增殖性糖尿病性视网膜病变患者血和玻璃体中的表达及其相关性研究.泸州:泸州医学院,2013
[10]Guex-Crosier Y,Behar-Cohen F.Diabetic retinopathy:New therapeuticpossibilities.RevueMedicaleSuisse,2015,11(456-457):101-107
[11]Mohammad Shamsul Ola,Mohd Imtiaz Nawaz,M Mairaj Siddiquei,et al.Recent advances inunderstanding the biochemical and molecular mechanism of diabetic retinopathy.J Diabetes and its Complications,2012,26(1):56-64
[12]Gille H,Kowalski J,Li B,et al.Analysis of biological effects and signaling properties of Flt-1(VEGFR-1)and KDR(VEGFR-2).Biol Chem,2007,276:3222-3230
[13]Sarbassov D D,Guertin D A,Ali S M,et al.Phosphorylation and regulation of Akt/PKB by the rictor-mTOR complex.Science,2005,307(5712):1098-10101
[14]Jacinto E,Facchinetti V,Liu D,et al.SIN1/MIP1 maintains rictorm TOR complex integrity and regulates Akt phosphorylation and substrate specificity.Cell,2006,127(1):125-137
[15]段俊国,金明,接传红.糖尿病视网膜病变中医防治指南.中国中医药现代远程教育,2011,9(4):154-155
[16]罗向霞,段俊国,李响.糖尿病视网膜病变阳虚病机的代谢组学物质基础.中国中医眼科杂志,2013,23(12):394