针刺对慢性高眼压后兔模型NGF-TrkA及AKT的影响
发布时间：2019-01-06 05:35
【摘要】：目的通过观察内源性NGF-TrkA及下游通路的分子基因转录特点,探讨针刺治疗对高眼压后兔视网膜神经节细胞的保护作用。方法 30只新西兰兔,采用随机数字表法随机将8只设为正常组,剩余22只造模。再次采用随机数字表法将造模成功后16只随机分为模型组、针刺组,每组8只兔。模型组与针刺组以复方卡波姆诱导建立慢性高眼压兔模型。造模2周后,针刺组开始给予毫针"合谷(LI4)、睛明(BL1)、球后(Ex-HN07)"穴治疗,每周3次,隔天1次,共治疗4周(12次);正常组和模型组不予治疗。治疗结束后,取3组兔眼球,行视网膜HE染色观测组织内形态学变化;采用RT-PCR法检测视网膜内神经生长因子信使核糖核酸(NGF mRNA)、神经生长因子受体酪氨酸激酶信使核糖核酸(TrkA mRNA)和神经营养素受体信使核糖核酸(p75 mRNA)的转录水平;以Western blotting法检测视网膜内NGF信号通路下游分子AKT(p-AKT)的表达水平。结果在治疗过程中,针刺组眼压均值波动在20.86~22.23 mm Hg之间,模型组眼压波动在23.82~25.49 mm Hg之间;造模后模型组、针刺组各时间点眼压与正常组比较,差异均有统计学意义(P0.05)。模型组视网膜整体结构有明显变化,内核层、外核层均排列混乱且两层之间边界不清,且内核层的双极细胞和外核层的视杆细胞、视锥细胞数量有所减少,神经节细胞层显示细胞核间距增大,数量有所减少。与模型组相比,针刺组内核层双极细胞数量稳定,内核层与外核层之间边界尚清,神经节细胞层细胞核间距减小,数量有所增加。视网膜内NGF mRNA、TrkA mRNA转录水平均增强,p75 mRNA转录水平降低,下游信号分子AKT表达针刺组较正常组增强,针刺组较模型组增强。结论①针刺作用于慢性高眼压后兔模型,能稳定眼压,可以使视网膜病理损害减轻,稳定神经节细胞数量,有效保护慢性高眼压后兔模型的RGC。②针刺可以提升NGF-TrkA在慢性高眼压兔模型视网膜组织内的转录水平,降低p75在慢性高眼压兔模型视网膜组织内的转录水平,提升下游信号分子AKT的磷酸化水平。
[Abstract]:Objective to investigate the protective effect of acupuncture on retinal ganglion cells (RGCs) after intraocular pressure (IOP) by observing the transcriptional characteristics of endogenous NGF-TrkA and its downstream pathway. Methods Thirty New Zealand rabbits were randomly divided into normal group (n = 8) and control group (n = 22). 16 rabbits were randomly divided into model group, acupuncture group and 8 rabbits in each group. The model group and acupuncture group were induced by compound carbomer to establish the model of chronic intraocular pressure. After 2 weeks of modeling, the acupuncture group was treated with "LI4", "BL1" and "Retrobulbar" (Ex-HN07), 3 times a week, once every other day, for 4 weeks (12 times), while the normal group and the model group were not treated. After the treatment, three groups of rabbit eyeballs were taken and the morphological changes in the tissues were observed by HE staining. The transcription levels of (NGF mRNA), receptor tyrosine kinase messenger ribonucleic acid (TrkA mRNA) and neurotrophin receptor messenger ribonucleic acid (p75 mRNA) in retina were measured by RT-PCR method. The expression of AKT (p-AKT) downstream of NGF signaling pathway in retina was detected by Western blotting method. Results in the course of treatment, the mean intraocular pressure fluctuated between 20.86 and 22.23 mm Hg in the acupuncture group and 23.82 卤25.49 mm Hg in the model group. There were significant differences in IOP between the model group and the acupuncture group at each time point compared with the normal group (P0.05). In the model group, the overall structure of the retina was obviously changed, the inner and outer nuclear layers were all arranged in disorder and the boundary between the two layers was not clear, and the number of cone cells in the inner layer and the outer nuclear layer was decreased. The ganglion cell layer showed an increase in nuclear spacing and a decrease in the number. Compared with the model group, the number of bipolar cells in the nuclear layer of the acupuncture group was stable, the boundary between the nuclear layer and the outer nuclear layer was clear, and the nucleus spacing of the ganglion cell layer decreased and the number increased. The transcription level of NGF mRNA,TrkA mRNA in retina was increased, the transcription level of p75 mRNA was decreased, the expression of downstream signal molecule AKT in acupuncture group was higher than that in normal group, and that in acupuncture group was higher than that in model group. Conclusion (1) Acupuncture can stabilize intraocular pressure, reduce retinal pathological damage and stabilize the number of ganglion cells after chronic high intraocular pressure in rabbits. The effective protection of RGC.2 acupuncture on chronic intraocular hypertension rabbit model can enhance the transcription level of NGF-TrkA in the retina of chronic high intraocular pressure rabbit model, and decrease the transcription level of p75 in the retina tissue of chronic high intraocular pressure rabbit model. The phosphorylation level of downstream signal molecule AKT was increased.
【作者单位】： 上海中医药大学门诊部;上海中医药大学科技实验中心;上海中医药大学附属曙光医院眼科;上海中医药大学附属曙光医院针灸科;
【基金】：上海市科委科研计划项目(14401971700) 上海中医药大学预算内项目(2014YSN29)
【分类号】：R245;R-332
，
本文编号：2402409
[Abstract]:Objective to investigate the protective effect of acupuncture on retinal ganglion cells (RGCs) after intraocular pressure (IOP) by observing the transcriptional characteristics of endogenous NGF-TrkA and its downstream pathway. Methods Thirty New Zealand rabbits were randomly divided into normal group (n = 8) and control group (n = 22). 16 rabbits were randomly divided into model group, acupuncture group and 8 rabbits in each group. The model group and acupuncture group were induced by compound carbomer to establish the model of chronic intraocular pressure. After 2 weeks of modeling, the acupuncture group was treated with "LI4", "BL1" and "Retrobulbar" (Ex-HN07), 3 times a week, once every other day, for 4 weeks (12 times), while the normal group and the model group were not treated. After the treatment, three groups of rabbit eyeballs were taken and the morphological changes in the tissues were observed by HE staining. The transcription levels of (NGF mRNA), receptor tyrosine kinase messenger ribonucleic acid (TrkA mRNA) and neurotrophin receptor messenger ribonucleic acid (p75 mRNA) in retina were measured by RT-PCR method. The expression of AKT (p-AKT) downstream of NGF signaling pathway in retina was detected by Western blotting method. Results in the course of treatment, the mean intraocular pressure fluctuated between 20.86 and 22.23 mm Hg in the acupuncture group and 23.82 卤25.49 mm Hg in the model group. There were significant differences in IOP between the model group and the acupuncture group at each time point compared with the normal group (P0.05). In the model group, the overall structure of the retina was obviously changed, the inner and outer nuclear layers were all arranged in disorder and the boundary between the two layers was not clear, and the number of cone cells in the inner layer and the outer nuclear layer was decreased. The ganglion cell layer showed an increase in nuclear spacing and a decrease in the number. Compared with the model group, the number of bipolar cells in the nuclear layer of the acupuncture group was stable, the boundary between the nuclear layer and the outer nuclear layer was clear, and the nucleus spacing of the ganglion cell layer decreased and the number increased. The transcription level of NGF mRNA,TrkA mRNA in retina was increased, the transcription level of p75 mRNA was decreased, the expression of downstream signal molecule AKT in acupuncture group was higher than that in normal group, and that in acupuncture group was higher than that in model group. Conclusion (1) Acupuncture can stabilize intraocular pressure, reduce retinal pathological damage and stabilize the number of ganglion cells after chronic high intraocular pressure in rabbits. The effective protection of RGC.2 acupuncture on chronic intraocular hypertension rabbit model can enhance the transcription level of NGF-TrkA in the retina of chronic high intraocular pressure rabbit model, and decrease the transcription level of p75 in the retina tissue of chronic high intraocular pressure rabbit model. The phosphorylation level of downstream signal molecule AKT was increased.
【作者单位】： 上海中医药大学门诊部;上海中医药大学科技实验中心;上海中医药大学附属曙光医院眼科;上海中医药大学附属曙光医院针灸科;
【基金】：上海市科委科研计划项目(14401971700) 上海中医药大学预算内项目(2014YSN29)
【分类号】：R245;R-332
，
本文编号：2402409
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