针刺对感光细胞凋亡模型鼠视网膜小胶质细胞活性的影响
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摘要
目的:观察针刺对感光细胞凋亡模型鼠视网膜小胶质细胞活性的影响。方法:选取6周龄的SD雄性大鼠,将N-甲基-N-亚硝基脲以50 mg/kg剂量一次性腹腔注射造成感光细胞凋亡动物模型。将大鼠随机分为正常组,模型组,维生素A组和针刺组,连续治疗14 d。治疗结束后采用HE染色观察视网膜形态改变; TUNEL法检测视网膜感光细胞凋亡情况;免疫荧光法检测小胶质细胞OX-42在视网膜中的表达。结果:与模型组比较,针刺组视网膜ONL厚度增加,并保留一定数量的感光细胞,针刺干预后TUNEL(+)凋亡细胞数量显著降低,差异具有统计学意义(P <0. 05)。与模型组和维生素A组比较,针刺组OX-42(+)细胞平均光密度降低(P <0. 05)且低于维生素A组(P <0. 05),提示小胶质细胞迁移、活化减少,针刺抑制小胶质细胞活性的效果优于维生素A组。结论:针刺对视网膜感光细胞具有保护作用,其可能通过抑制视网膜小胶质细胞的活化、迁移,进而减少视网膜感光细胞的凋亡。
Objective:To observe the effect of acupuncture on photoreceptor cells apoptosis,especially on the activity of retinal microglia in rats. Methods:MUN was injected with 50 mg/kg to abdominal cavity of SD rats with ages of 6 weeks to induce animal models of photoreceptor cells apoptosis. The rats were divided into normal group,model group,vitamin A group,acupuncture group and treated continuously for 14 days. HE staining was used to observe the changes of retinal tissue morphology. TUNEL method was used to detect the apoptosis of photoreceptor cells. The immunofluorescence was used to detect the expression of retinal microglia OX-42 in the retina. Results:Compared with the model group,the retinal ONL thickness in the acupuncture group increased and a certain amount of photoreceptor cells remained. After acupuncture intervention,the number of apoptotic cells in TUNEL( +) decreased significantly(P < 0. 05). Compared with the model group and the vitamin A group,the average optical density of OX-42( +) decreased in acupuncture group( P < 0. 05) and the average optical density of acupuncture group was lower than that of vitamin A group( P < 0. 05). It is suggested the decrease in microglia migration and activation and the effect of acupuncture on the activity of retinal microglia is better than that of vitamin A group. Conclusion:Acupuncture has a protective effect on retinal photoreceptor cells,which may reduce the apoptosis of retinal photoreceptor cells by inhibiting the migration and activation of microglia into retina.
Objective:To observe the effect of acupuncture on photoreceptor cells apoptosis,especially on the activity of retinal microglia in rats. Methods:MUN was injected with 50 mg/kg to abdominal cavity of SD rats with ages of 6 weeks to induce animal models of photoreceptor cells apoptosis. The rats were divided into normal group,model group,vitamin A group,acupuncture group and treated continuously for 14 days. HE staining was used to observe the changes of retinal tissue morphology. TUNEL method was used to detect the apoptosis of photoreceptor cells. The immunofluorescence was used to detect the expression of retinal microglia OX-42 in the retina. Results:Compared with the model group,the retinal ONL thickness in the acupuncture group increased and a certain amount of photoreceptor cells remained. After acupuncture intervention,the number of apoptotic cells in TUNEL( +) decreased significantly(P < 0. 05). Compared with the model group and the vitamin A group,the average optical density of OX-42( +) decreased in acupuncture group( P < 0. 05) and the average optical density of acupuncture group was lower than that of vitamin A group( P < 0. 05). It is suggested the decrease in microglia migration and activation and the effect of acupuncture on the activity of retinal microglia is better than that of vitamin A group. Conclusion:Acupuncture has a protective effect on retinal photoreceptor cells,which may reduce the apoptosis of retinal photoreceptor cells by inhibiting the migration and activation of microglia into retina.
引文
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[13]Shijie J,Jun K,Tstsuya M,et al.Interferon-βis neuroprotective against the toxicity induced by activated microglia[J].Brain Resarch,2007,179(7):140-146.
[14]EBERT S,WALCZAK Y,REME C,et al.Microglial activation and transcriptomic changes in the blue light-ex-posed mouse retina[J].Adv Exp Med Biol,2012,723:619-632.
[15]邵敬芝,彭广华.干细胞移植治疗视网膜变性病的研究进展[J].眼科新展,2016,36(2):118.
[16]曾庆广,左志高.针刺新明穴治疗外伤性视神经萎缩180例疗效分析[J].武警医学,1998,9(4):213.
[2]王莹,张露,叶河江,等.针刺对感光细胞凋亡大鼠视网膜组织中谷氨酸含量的影响[J].中医眼耳鼻喉杂志,2011,1(4):209-211.
[3]华兴邦.大鼠穴位图谱研制[J].实验动物与动物实验,1991(1):765.
[4]周媛,曹霞.视网膜神经胶质细胞及其分泌的细胞因子与视网膜病变[J].细胞与分子免疫学杂志,2012,28(9):996-998.
[5]Minghetti L.Role of inflammation in neurodegenerative diseases[J].Curr Opin Neurol,2005,18:315-321.
[6]Santiago AR,Baptista FI,Santos PF,et al.Role of microglial adeonsine A(2A)receptors in retinal and brain neurodegenerative diseases[J].Mediators Inflame,2014(2):465-694.
[7]Santos A M,Martín-Oliva D,Ferrer-Martín R M,et al.Microglial response to light-induced photoreceptor degeneration in the mouse retina[J].Journal of Comparative Neurology,2010,518(4):477.
[8]Socodato R1,Portugal CC,Domith I,et al.c-Src function is necessary and sufficient for triggering microglial cell activation[J].Glia,2015,63(3):497-511.
[9]Kawabori M,Yenari MA.The role of the microglia in acute CNS injury[J].Metab Brain Dis,2015,30(2):381-392.
[10]Lioba Horstmann,Sandra Kuehn,Xiomara Pedreiturria,et al.Microglia response in retina and optic nerve in chronic experimental autoimmune encephalomyelitis[J].Neuroimmunology,2016,298:32-41.
[11]Lian Zhao,Matthew K Zabel,Xu Wang,et al.Microglial phagocytosis of living photoreceptors contributes to inherited retinal degeneration[J].EBMO Molecular Medicine,2015,7(9):1179-1197.
[12]Rivera JC,Sitaras N,Noueihed B,et al.Microglia and interleukin-1βin ischemic retinopathy elicit microvascular degeneration through neuronal semaphorin-3A[J].Arterioscler Thromb Vasc Biol,2013,33(8):1881-1891.
[13]Shijie J,Jun K,Tstsuya M,et al.Interferon-βis neuroprotective against the toxicity induced by activated microglia[J].Brain Resarch,2007,179(7):140-146.
[14]EBERT S,WALCZAK Y,REME C,et al.Microglial activation and transcriptomic changes in the blue light-ex-posed mouse retina[J].Adv Exp Med Biol,2012,723:619-632.
[15]邵敬芝,彭广华.干细胞移植治疗视网膜变性病的研究进展[J].眼科新展,2016,36(2):118.
[16]曾庆广,左志高.针刺新明穴治疗外伤性视神经萎缩180例疗效分析[J].武警医学,1998,9(4):213.