阿柏西普对大鼠视网膜Müller细胞STAT3表达的影响
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摘要
目的:探讨阿柏西普对大鼠视网膜Müller细胞信号转导及转录活化因子3(STAT3)表达的影响。方法:采用不同浓度的阿柏西普100μL(加药浓度分别为400、200、100pg/mL)大鼠永生化视网膜Müller细胞24、48h后,MTT法检测细胞增殖,流式细胞仪检测细胞凋亡,Transwell小室法检测细胞侵袭,Western-blot法检测AKT、STAT3蛋白表达。结果:随着阿柏西普浓度的升高,Müller细胞的增殖活性下降(P<0.05)。处理48h后,随着阿柏西普浓度的升高,Müller细胞的凋亡率逐渐上升,Müller细胞的侵袭穿透指数逐渐降低(P<0.05)。转染48h后,随着阿柏西普浓度的升高,Müller细胞的AKT蛋白相对表达量无显著变化(P>0.05),STAT3蛋白相对表达量逐渐降低(P<0.05)。结论:阿柏西普可抑制大鼠视网膜Müller细胞STAT3蛋白表达,从而抑制细胞增殖与侵袭性,促进细胞凋亡。
·AIM: To investigate the effect of aflibercept on the expression of signal transducers and activators of transcription 3(STAT3) in rat retinal Müller cells. ·METHODS: The rat retinal Müller cells treated with different concentrations of Aflibercept 100 μL(diluted concentrations of 400, 200, 100 pg/mL, respectively). MTT assay were used to detect cell proliferation, flow cytometry. Apoptosis were detected by the instrument, cell invasion were detected by transwell chamber method, and protein(AKT, STAT3, GAPDH) expression were detected by Western-blot method.·RESULTS: The proliferation activity of Müller cells were decreased with the increased of aflibercept concentration, and compared the difference were statistically significant(P<0.05). After treatment for 48 h, the apoptotic rate of Müller cells were gradually increased with the increased of aflibercept concentration, and the invasion and penetration index of Müller cells gradually were decreased, and compared the difference were statistically significant(P<0.05). After 48 h of transfection, the relative expression of AKT protein in Müller cells were not change significantly with the increased of Aflibercept concentration(P>0.05), and the relative expression of STAT3 protein decreased gradually, and compared the difference were statistically significant(P<0.05).·CONCLUSION: Aflibercept can inhibit the expression of STAT3 protein in rat retinal Müller cells, thereby inhibit cell proliferation and invasion and promote apoptosis.
·AIM: To investigate the effect of aflibercept on the expression of signal transducers and activators of transcription 3(STAT3) in rat retinal Müller cells. ·METHODS: The rat retinal Müller cells treated with different concentrations of Aflibercept 100 μL(diluted concentrations of 400, 200, 100 pg/mL, respectively). MTT assay were used to detect cell proliferation, flow cytometry. Apoptosis were detected by the instrument, cell invasion were detected by transwell chamber method, and protein(AKT, STAT3, GAPDH) expression were detected by Western-blot method.·RESULTS: The proliferation activity of Müller cells were decreased with the increased of aflibercept concentration, and compared the difference were statistically significant(P<0.05). After treatment for 48 h, the apoptotic rate of Müller cells were gradually increased with the increased of aflibercept concentration, and the invasion and penetration index of Müller cells gradually were decreased, and compared the difference were statistically significant(P<0.05). After 48 h of transfection, the relative expression of AKT protein in Müller cells were not change significantly with the increased of Aflibercept concentration(P>0.05), and the relative expression of STAT3 protein decreased gradually, and compared the difference were statistically significant(P<0.05).·CONCLUSION: Aflibercept can inhibit the expression of STAT3 protein in rat retinal Müller cells, thereby inhibit cell proliferation and invasion and promote apoptosis.
引文
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2金姬,卢一,冯佳,等.视网膜Müller细胞条件性基因敲除血管内皮生长因子对氧诱导视网膜病变小鼠的影响.中华眼底病杂志2017; 33(5):508-512
3 Ranski AH, Kramer AC, Morgan GW,et al. Characterization of retinal regeneration in adult zebrafish following multiple rounds of phototoxic lesion. Peer J 2018;20(6):e5646
4 Pozios I, Kn?sel T, Zhao Y,et al. Expression of phosphorylated estrogen receptor beta is an independent negative prognostic factor for pancreatic ductal adenocarcinoma. J Cancer Res Clin Oncol 2018;144(10):1887-1897
5姚晓倩,方媛,陈君毅.调控Müller细胞增殖分化的因子及信号通路的研究.中国眼耳鼻喉科杂志 2017; 17(6):431-435
6 L?cken H, Clamor C, Müller K. Napabucasin and Related Heterocycle-Fused Naphthoquinones as STAT3 Inhibitors with Antiproliferative Activity against Cancer Cells. J Nat Prod 2018; 81(7):1636-1644
7 Shin SY, Müller AK, Verma N,et al. Systems modelling of the EGFR-PYK2-c-Met interaction network predicts and prioritizes synergistic drug combinations for triple-negative breast cancer. PLoS Comput Biol 2018; 14(6):e1006192
8杨雄里.视网膜神经信号的调制和视网膜疾病.复旦学报(医学版) 2017; 44(6):693-698
9章淑杰,张荣,张雪瑾,等.人Müller细胞来源的外泌体对脂多糖刺激视网膜色素上皮细胞产生炎症因子的影响.中国眼耳鼻喉科杂志 2017; 17(5):323-327
10 Ziegler PK, Bollrath J, Pallangyo CK,et al. Mitophagy in Intestinal Epithelial Cells Triggers Adaptive Immunity during Tumorigenesis. Cell 2018; 174(1):88-101
11 Arndt L, Dokas J, Gericke M,et al. Tribbles homolog 1 deficiency modulates function and polarization of murine bone marrow-derived macrophages. J Biol Chem 2018; 293(29):11527-11536
12丛小杰,肖宗宇,李文辉,等.Müller细胞在视网膜病变发生发展中作用的研究进展.山东医药 2017; 57(37):107-109
13 Ham S, Lima LG, Chai EPZ,et al. Breast Cancer-Derived Exosomes Alter Macrophage Polarization via gp130/STAT3 Signaling. Front Immunol 2018; 8(9):871
14 Ren JL, Yu QX, Liang WC,et al. Green tea extract attenuates LPS-induced retinal inflammation in rats. Sci Rep 2018; 8(1):429
15汪晓磊,马建民,孟照洋,等.急性高眼压模型中p75NTR抑制剂对视网膜神经节细胞作用的研究.首都医科大学学报 2017; 38(1):24-28
16 Eichler W,Savkovic-Cvijic H,Burger S,et al.Muller Cell-Derived PEDF Mediates Neuroprotection via STAT3 Activation. Cell Physiol Biochem 2017; 44(4):1411-1424
17 Ahmad R, Kumar B, Chen Z,et al. Loss of claudin-3 expression induces IL6/gp130/Stat3 signaling to promote colon cancer malignancy by hyperactivating Wnt/β-cateni signaling. Oncogene 2017; 36(47):6592-6604
18田敏,吴进川,何薇,等.叔丁基对苯二酚对高糖培养视网膜Müller细胞核因子E2相关因子2、血红素氧合酶1和磷脂酰肌醇-3激酶表达的影响.中华眼底病杂志 2018; 34(4):382-387
19 Vargas A, Kim HS, Baral E,et al. Protective effect of clusterin on rod photoreceptor in rat model of retinitis pigmentosa. PLoS One 2017; 12(8):e0182389
20梁汇珉,刘学政.脑源性神经营养因子(BDNF)对高糖环境下视网膜Müller细胞的保护作用.眼科新进展 2018; 38(8):714-718
21 Schwerd T, Twigg SRF, Aschenbrenner D,et al. A biallelic mutation in IL6ST encoding the GP130 co-receptor causes immunodeficiency and craniosynostosis. J Exp Med 2017; 214(9):2547-2562
22余爱华,柯敏,田朕,等.P58IPK基因在视网膜Müller细胞内质网应激PERK通道中的作用.武汉大学学报(医学版) 2018; 39(2):213-218
23 Zhu S, Luo H, Liu H,et al. p38MAPK plays a critical role in induction of a pro-inflammatory phenotype of retinal Müller cells following Zika virus infection. Antiviral Res 2017; 9(145):70-81
24杨子建,程瑜,姚慧萍,等.腺苷受体拮抗剂对加压培养的视网膜Müller细胞钾离子通道的调控作用.中华实验眼科杂志 2018; 36(8):590-594
25 Armento A, Ilina EI, Kaoma T,et al. Carboxypeptidase E transmits its anti-migratory function in glioma cells via transcriptional regulation of cell architecture and motility regulating factors. Int J Oncol 2017; 51(2):702-714
2金姬,卢一,冯佳,等.视网膜Müller细胞条件性基因敲除血管内皮生长因子对氧诱导视网膜病变小鼠的影响.中华眼底病杂志2017; 33(5):508-512
3 Ranski AH, Kramer AC, Morgan GW,et al. Characterization of retinal regeneration in adult zebrafish following multiple rounds of phototoxic lesion. Peer J 2018;20(6):e5646
4 Pozios I, Kn?sel T, Zhao Y,et al. Expression of phosphorylated estrogen receptor beta is an independent negative prognostic factor for pancreatic ductal adenocarcinoma. J Cancer Res Clin Oncol 2018;144(10):1887-1897
5姚晓倩,方媛,陈君毅.调控Müller细胞增殖分化的因子及信号通路的研究.中国眼耳鼻喉科杂志 2017; 17(6):431-435
6 L?cken H, Clamor C, Müller K. Napabucasin and Related Heterocycle-Fused Naphthoquinones as STAT3 Inhibitors with Antiproliferative Activity against Cancer Cells. J Nat Prod 2018; 81(7):1636-1644
7 Shin SY, Müller AK, Verma N,et al. Systems modelling of the EGFR-PYK2-c-Met interaction network predicts and prioritizes synergistic drug combinations for triple-negative breast cancer. PLoS Comput Biol 2018; 14(6):e1006192
8杨雄里.视网膜神经信号的调制和视网膜疾病.复旦学报(医学版) 2017; 44(6):693-698
9章淑杰,张荣,张雪瑾,等.人Müller细胞来源的外泌体对脂多糖刺激视网膜色素上皮细胞产生炎症因子的影响.中国眼耳鼻喉科杂志 2017; 17(5):323-327
10 Ziegler PK, Bollrath J, Pallangyo CK,et al. Mitophagy in Intestinal Epithelial Cells Triggers Adaptive Immunity during Tumorigenesis. Cell 2018; 174(1):88-101
11 Arndt L, Dokas J, Gericke M,et al. Tribbles homolog 1 deficiency modulates function and polarization of murine bone marrow-derived macrophages. J Biol Chem 2018; 293(29):11527-11536
12丛小杰,肖宗宇,李文辉,等.Müller细胞在视网膜病变发生发展中作用的研究进展.山东医药 2017; 57(37):107-109
13 Ham S, Lima LG, Chai EPZ,et al. Breast Cancer-Derived Exosomes Alter Macrophage Polarization via gp130/STAT3 Signaling. Front Immunol 2018; 8(9):871
14 Ren JL, Yu QX, Liang WC,et al. Green tea extract attenuates LPS-induced retinal inflammation in rats. Sci Rep 2018; 8(1):429
15汪晓磊,马建民,孟照洋,等.急性高眼压模型中p75NTR抑制剂对视网膜神经节细胞作用的研究.首都医科大学学报 2017; 38(1):24-28
16 Eichler W,Savkovic-Cvijic H,Burger S,et al.Muller Cell-Derived PEDF Mediates Neuroprotection via STAT3 Activation. Cell Physiol Biochem 2017; 44(4):1411-1424
17 Ahmad R, Kumar B, Chen Z,et al. Loss of claudin-3 expression induces IL6/gp130/Stat3 signaling to promote colon cancer malignancy by hyperactivating Wnt/β-cateni signaling. Oncogene 2017; 36(47):6592-6604
18田敏,吴进川,何薇,等.叔丁基对苯二酚对高糖培养视网膜Müller细胞核因子E2相关因子2、血红素氧合酶1和磷脂酰肌醇-3激酶表达的影响.中华眼底病杂志 2018; 34(4):382-387
19 Vargas A, Kim HS, Baral E,et al. Protective effect of clusterin on rod photoreceptor in rat model of retinitis pigmentosa. PLoS One 2017; 12(8):e0182389
20梁汇珉,刘学政.脑源性神经营养因子(BDNF)对高糖环境下视网膜Müller细胞的保护作用.眼科新进展 2018; 38(8):714-718
21 Schwerd T, Twigg SRF, Aschenbrenner D,et al. A biallelic mutation in IL6ST encoding the GP130 co-receptor causes immunodeficiency and craniosynostosis. J Exp Med 2017; 214(9):2547-2562
22余爱华,柯敏,田朕,等.P58IPK基因在视网膜Müller细胞内质网应激PERK通道中的作用.武汉大学学报(医学版) 2018; 39(2):213-218
23 Zhu S, Luo H, Liu H,et al. p38MAPK plays a critical role in induction of a pro-inflammatory phenotype of retinal Müller cells following Zika virus infection. Antiviral Res 2017; 9(145):70-81
24杨子建,程瑜,姚慧萍,等.腺苷受体拮抗剂对加压培养的视网膜Müller细胞钾离子通道的调控作用.中华实验眼科杂志 2018; 36(8):590-594
25 Armento A, Ilina EI, Kaoma T,et al. Carboxypeptidase E transmits its anti-migratory function in glioma cells via transcriptional regulation of cell architecture and motility regulating factors. Int J Oncol 2017; 51(2):702-714