养血补肾方对实验性近视眼视网膜p-Stat3活性蛋白的表达影响
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摘要
目的观察养血补肾方对实验性近视眼视网膜p-Stat3表达变化的影响,为治疗近视眼提供相关实验依据。方法取2周龄健康的豚鼠,用半透明眼罩行右眼遮盖60 d制备形觉剥夺性近视模型,将造模成功的豚鼠随机分为对照组和治疗组,并测量豚鼠造模前后双眼的屈光度(D)和眼轴(L)。治疗组给予养血补肾方水煎液(生药量12.7 g/kg)连续灌胃21 d,每日一次,对照组不进行干预。采用HE染色法观察视网膜组织形态学变化,采用免疫组化、Western-blot等方法检测视网膜p-Stat3的表达变化。结果半透明眼罩遮盖豚鼠右眼60 d后成功制备实验眼剥夺性近视模型,与自身对照眼相比屈光度明显增加,剥夺前屈光度为(3.924±3.392)D,剥夺后(-2.583±2.954)D,P=0.001,差异有统计学意义。造模前右眼(形觉剥夺眼)眼轴平均长度为(7.557±0.176)mm,造模后右眼眼轴平均长度为(8.278±0.250)mm,眼轴长度明显增加,P=0.01,差异具有统计学意义。与对照组相比,治疗组灌胃21 d后,p-Stat3在未遮盖豚鼠眼视网膜中有微弱表达,遮盖后p-Stat3表达相比遮盖前上调,差异无统计学意义(P>0.05)。结论 p-Stat3信号通路均参与了实验性近视的形成和发展,而养血补肾方对视网膜p-Stat3蛋白的表达变化无影响。
OBJECTIVE To observe the influences of Yangxue Bushen formula on the expression of retinal p-Stat3 in experimental myopia tissue, and to provide experimental support for this formula on high myopia. METHODS Two-weeks-old guinea pigs were used to prepare models of form deprivation myopia by covering right eye with translucent goggles for 60 days. The successful models were randomly divided into control and treatment group, and the eye diopter(D)and axial length(L) before and after molding were measured. The treatment group received Yangxu Bushen formula(crude drug content in 12.7 g/kg) i.g for 21 d, once a day; the control group was not treated by any intervention. The immunohistochemistry and Western-blot were used to detect the retinal p-Stat3 expression. RESULTS The models of experimental eye deprivation myopia were successfully made after 60 d of monocular covering with translucent goggles. Compared with the result before covering, diopter increased significantly: the diopter before deprivation was(3.924 ±3.392) D, and it after deprivation was(-2.583 ±2.954) D. The difference was of statistical significance(P<0.05). Axial average length of the right eye before modeling(form deprivation eyes) was(7.557 ±0.176) mm; after modeling,the average length was(8.278±0.250) mm. The axial length increased significantly(P <0.05).Compared with the control group, weak expression of p-Stat3 was observed in retina of uncovered eye while p-Stat3 expression increased after covering in treatment group. However, there was no statistical difference on p-Stat3 expression between the treatment group and the control group(P >0.05). CONCLUSIONS The signaling pathway of p-Stat3 played a part in occurrence and development of form deprivation myopia. The Yangxue Bushen formula showed no influence on expression of retinal p-Stat3 protein.
OBJECTIVE To observe the influences of Yangxue Bushen formula on the expression of retinal p-Stat3 in experimental myopia tissue, and to provide experimental support for this formula on high myopia. METHODS Two-weeks-old guinea pigs were used to prepare models of form deprivation myopia by covering right eye with translucent goggles for 60 days. The successful models were randomly divided into control and treatment group, and the eye diopter(D)and axial length(L) before and after molding were measured. The treatment group received Yangxu Bushen formula(crude drug content in 12.7 g/kg) i.g for 21 d, once a day; the control group was not treated by any intervention. The immunohistochemistry and Western-blot were used to detect the retinal p-Stat3 expression. RESULTS The models of experimental eye deprivation myopia were successfully made after 60 d of monocular covering with translucent goggles. Compared with the result before covering, diopter increased significantly: the diopter before deprivation was(3.924 ±3.392) D, and it after deprivation was(-2.583 ±2.954) D. The difference was of statistical significance(P<0.05). Axial average length of the right eye before modeling(form deprivation eyes) was(7.557 ±0.176) mm; after modeling,the average length was(8.278±0.250) mm. The axial length increased significantly(P <0.05).Compared with the control group, weak expression of p-Stat3 was observed in retina of uncovered eye while p-Stat3 expression increased after covering in treatment group. However, there was no statistical difference on p-Stat3 expression between the treatment group and the control group(P >0.05). CONCLUSIONS The signaling pathway of p-Stat3 played a part in occurrence and development of form deprivation myopia. The Yangxue Bushen formula showed no influence on expression of retinal p-Stat3 protein.
引文
[1]侯静梅.养血补肾方对实验性高度近视巩膜细胞凋亡的保护作用[D].西宁:青海大学,2015.
[2] SAMUEL SMZ, MU GL, ARIHIRO A, et a1. STAT3 activation in response to growth factors cytokines panicipates in retina pre-cursor proliferation[J].Exp Eye Res,2005,81(1):103-l15.
[3] OZAWA Y, NAKAO K, KURIHARA T, et al.Roles of STAT3/SOCS3pathway in regulating the visual function and ubiquitin-proteasomedependent degradation of rhodopsin during retinal inflammation[J].J Biol Chem, 2008,283(36):24561-24570.
[4] KWAK HB, SUN HM, HA H, et al. AG490, a Jak2-specific inhibitor, induces osteoclast survival by activating the Akt and ERK signaling pathways[J].Mol Cells,2008,26(5):436-442.
[5]朱子诚,张金嵩.豚鼠形觉剥夺性近视中视网膜stat3活性蛋白的表达[J].安徽医学,2009,30(8):860-862.
[6]王江辉.多波长激光诱导豚鼠形觉剥夺性高度近视脉络膜新生血管模型建立的研究[D].西宁:青海大学,2012.
[7] LEONARD WJ, O SHEA JJ. Jaks and STATs:biological implications[J].Annu Rev Immunol,1998,16(1):293-322.
[8] OSHEA JJ, JAKS, STATS.Cytokine signaling transduction and immunoregulation:are we there yet?[J].Immunity,1997,7(1):1-11.
[9] ZHANG SS, WEI JY, LI C, et al. Expression and activation of STAT proteins during mouse retina development[J].Exp Eye Res,2003,76(4):421-431.
[10]庞国龙,孙河.Stat3在眼科领域的研究现状和前景展望[J],中国中医眼科杂志,2013,23(5):386-388.
[11]王静波,郑新青.四物五子汤中微量元素的测定[J].中国中医眼科杂志,1993,3(2):93.
[12] PETERSON WM, WANG Q, TZEKOVA R, et a1. Ciliary neumtmphic factor and stress stimuli activate the Jak-Stat pathway inretinal neurons and glia[J].J Neumsci,2000,20(11):4081-4090.
[13] ZHANG SS, WEI JY, LI C, et al. Expression and activation of STAT proteins during mouse retina development[J].Exp Eye Res,2003,76(4):421-431.
[14]胡诞宁,STEVEN A,MCCORMICK.视网膜色素上皮-脉络膜在近视发病中的作用[J].眼视光学杂志,2000,2(4):197-200.
[15] XIE TX, WEI D,LIU M, et al.Stat3 activation regulates the expression of matrix metalloproteinase-2 and tumor invasion and metastasis[J].Oncogene,2004,23(20):3550-3560.
[16] RIVAT C, RODRIGUES S, BRUYNEEL E, et a1. Implication of stat3 signaling in human colonic Cancer cells during intestinal trefoil factor3(TFF3)and vascular endothelial growth factor mediated cellular invasion and tumor growth[J]. Cancer Res,2005,65(1):195-202.
[2] SAMUEL SMZ, MU GL, ARIHIRO A, et a1. STAT3 activation in response to growth factors cytokines panicipates in retina pre-cursor proliferation[J].Exp Eye Res,2005,81(1):103-l15.
[3] OZAWA Y, NAKAO K, KURIHARA T, et al.Roles of STAT3/SOCS3pathway in regulating the visual function and ubiquitin-proteasomedependent degradation of rhodopsin during retinal inflammation[J].J Biol Chem, 2008,283(36):24561-24570.
[4] KWAK HB, SUN HM, HA H, et al. AG490, a Jak2-specific inhibitor, induces osteoclast survival by activating the Akt and ERK signaling pathways[J].Mol Cells,2008,26(5):436-442.
[5]朱子诚,张金嵩.豚鼠形觉剥夺性近视中视网膜stat3活性蛋白的表达[J].安徽医学,2009,30(8):860-862.
[6]王江辉.多波长激光诱导豚鼠形觉剥夺性高度近视脉络膜新生血管模型建立的研究[D].西宁:青海大学,2012.
[7] LEONARD WJ, O SHEA JJ. Jaks and STATs:biological implications[J].Annu Rev Immunol,1998,16(1):293-322.
[8] OSHEA JJ, JAKS, STATS.Cytokine signaling transduction and immunoregulation:are we there yet?[J].Immunity,1997,7(1):1-11.
[9] ZHANG SS, WEI JY, LI C, et al. Expression and activation of STAT proteins during mouse retina development[J].Exp Eye Res,2003,76(4):421-431.
[10]庞国龙,孙河.Stat3在眼科领域的研究现状和前景展望[J],中国中医眼科杂志,2013,23(5):386-388.
[11]王静波,郑新青.四物五子汤中微量元素的测定[J].中国中医眼科杂志,1993,3(2):93.
[12] PETERSON WM, WANG Q, TZEKOVA R, et a1. Ciliary neumtmphic factor and stress stimuli activate the Jak-Stat pathway inretinal neurons and glia[J].J Neumsci,2000,20(11):4081-4090.
[13] ZHANG SS, WEI JY, LI C, et al. Expression and activation of STAT proteins during mouse retina development[J].Exp Eye Res,2003,76(4):421-431.
[14]胡诞宁,STEVEN A,MCCORMICK.视网膜色素上皮-脉络膜在近视发病中的作用[J].眼视光学杂志,2000,2(4):197-200.
[15] XIE TX, WEI D,LIU M, et al.Stat3 activation regulates the expression of matrix metalloproteinase-2 and tumor invasion and metastasis[J].Oncogene,2004,23(20):3550-3560.
[16] RIVAT C, RODRIGUES S, BRUYNEEL E, et a1. Implication of stat3 signaling in human colonic Cancer cells during intestinal trefoil factor3(TFF3)and vascular endothelial growth factor mediated cellular invasion and tumor growth[J]. Cancer Res,2005,65(1):195-202.