針刺對(duì)慢性高眼壓后兔模型NGF-TrkA及AKT的影響
發(fā)布時(shí)間：2019-01-06 05:35
【摘要】：目的通過觀察內(nèi)源性NGF-TrkA及下游通路的分子基因轉(zhuǎn)錄特點(diǎn),探討針刺治療對(duì)高眼壓后兔視網(wǎng)膜神經(jīng)節(jié)細(xì)胞的保護(hù)作用。方法 30只新西蘭兔,采用隨機(jī)數(shù)字表法隨機(jī)將8只設(shè)為正常組,剩余22只造模。再次采用隨機(jī)數(shù)字表法將造模成功后16只隨機(jī)分為模型組、針刺組,每組8只兔。模型組與針刺組以復(fù)方卡波姆誘導(dǎo)建立慢性高眼壓兔模型。造模2周后,針刺組開始給予毫針"合谷(LI4)、睛明(BL1)、球后(Ex-HN07)"穴治療,每周3次,隔天1次,共治療4周(12次);正常組和模型組不予治療。治療結(jié)束后,取3組兔眼球,行視網(wǎng)膜HE染色觀測(cè)組織內(nèi)形態(tài)學(xué)變化;采用RT-PCR法檢測(cè)視網(wǎng)膜內(nèi)神經(jīng)生長(zhǎng)因子信使核糖核酸(NGF mRNA)、神經(jīng)生長(zhǎng)因子受體酪氨酸激酶信使核糖核酸(TrkA mRNA)和神經(jīng)營(yíng)養(yǎng)素受體信使核糖核酸(p75 mRNA)的轉(zhuǎn)錄水平;以Western blotting法檢測(cè)視網(wǎng)膜內(nèi)NGF信號(hào)通路下游分子AKT(p-AKT)的表達(dá)水平。結(jié)果在治療過程中,針刺組眼壓均值波動(dòng)在20.86~22.23 mm Hg之間,模型組眼壓波動(dòng)在23.82~25.49 mm Hg之間;造模后模型組、針刺組各時(shí)間點(diǎn)眼壓與正常組比較,差異均有統(tǒng)計(jì)學(xué)意義(P0.05)。模型組視網(wǎng)膜整體結(jié)構(gòu)有明顯變化,內(nèi)核層、外核層均排列混亂且兩層之間邊界不清,且內(nèi)核層的雙極細(xì)胞和外核層的視桿細(xì)胞、視錐細(xì)胞數(shù)量有所減少,神經(jīng)節(jié)細(xì)胞層顯示細(xì)胞核間距增大,數(shù)量有所減少。與模型組相比,針刺組內(nèi)核層雙極細(xì)胞數(shù)量穩(wěn)定,內(nèi)核層與外核層之間邊界尚清,神經(jīng)節(jié)細(xì)胞層細(xì)胞核間距減小,數(shù)量有所增加。視網(wǎng)膜內(nèi)NGF mRNA、TrkA mRNA轉(zhuǎn)錄水平均增強(qiáng),p75 mRNA轉(zhuǎn)錄水平降低,下游信號(hào)分子AKT表達(dá)針刺組較正常組增強(qiáng),針刺組較模型組增強(qiáng)。結(jié)論①針刺作用于慢性高眼壓后兔模型,能穩(wěn)定眼壓,可以使視網(wǎng)膜病理?yè)p害減輕,穩(wěn)定神經(jīng)節(jié)細(xì)胞數(shù)量,有效保護(hù)慢性高眼壓后兔模型的RGC。②針刺可以提升NGF-TrkA在慢性高眼壓兔模型視網(wǎng)膜組織內(nèi)的轉(zhuǎn)錄水平,降低p75在慢性高眼壓兔模型視網(wǎng)膜組織內(nèi)的轉(zhuǎn)錄水平,提升下游信號(hào)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 鹵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.
【作者單位】： 上海中醫(yī)藥大學(xué)門診部;上海中醫(yī)藥大學(xué)科技實(shí)驗(yàn)中心;上海中醫(yī)藥大學(xué)附屬曙光醫(yī)院眼科;上海中醫(yī)藥大學(xué)附屬曙光醫(yī)院針灸科;
【基金】：上海市科委科研計(jì)劃項(xiàng)目(14401971700) 上海中醫(yī)藥大學(xué)預(yù)算內(nèi)項(xiàng)目(2014YSN29)
【分類號(hào)】：R245;R-332
，
本文編號(hào)：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 鹵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.
【作者單位】： 上海中醫(yī)藥大學(xué)門診部;上海中醫(yī)藥大學(xué)科技實(shí)驗(yàn)中心;上海中醫(yī)藥大學(xué)附屬曙光醫(yī)院眼科;上海中醫(yī)藥大學(xué)附屬曙光醫(yī)院針灸科;
【基金】：上海市科委科研計(jì)劃項(xiàng)目(14401971700) 上海中醫(yī)藥大學(xué)預(yù)算內(nèi)項(xiàng)目(2014YSN29)
【分類號(hào)】：R245;R-332
，
本文編號(hào)：2402409
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