菊睛丸改善D-半乳糖致亚急性衰老小鼠视网膜损伤效应评价与作用机制研究

Effect Evaluation and Mechanism of Jujing Pills in Improving Retinal Injury in Subacute Aging Mice Induced by D-galactose

  • 摘要:
      目的  评价菊睛丸改善D-半乳糖致亚急性衰老小鼠视网膜损伤效应并探索作用机制。
      方法  采用小鼠皮下注射D-半乳糖构建亚急性衰老模型, 分别给予西药阳性药AREDS2、中药阳性药杞菊地黄丸、菊睛丸水提取物低、高剂量组和菊睛丸50%乙醇提取物低、高剂量组。通过光学相干断层成像检查(OCT)检测小鼠视网膜及眼底情况, HE染色法检测视网膜细胞损伤程度, TUNEL染色法检测视网膜细胞凋亡情况, 试剂盒检测氧化应激因子表达, Western blot检测小鼠视网膜凋亡和氧化应激通路相关蛋白表达情况。
      结果  菊睛丸提取物显著增加模型小鼠视网膜厚度(P < 0.05), 改善视网膜损伤并抑制视网膜细胞凋亡(P < 0.05)。同时, 菊睛丸提取物下调Cleaved Caspase-3与Cleaved Caspase-9凋亡通路蛋白表达(P < 0.05), 上调Bcl-2/Bax蛋白表达比值(P < 0.05)。菊睛丸提取物显著提高抗氧化因子SOD与GSH等表达(P < 0.05), 减少氧化产物MDA产生(P < 0.05), 显著上调抗氧化蛋白Nrf2与HO-1的表达(P < 0.05)。菊睛丸水提取物和50%乙醇提取物有相似的作用趋势。
      结论  菊睛丸改善D-半乳糖亚急性衰老模型小鼠视网膜损伤, 其作用途径与抑制视网膜细胞凋亡并改善氧化应激有关。

     

    Abstract:
      OBJECTIVE  To evaluate the effect of Jujing Pills on retinal injury in subacute aging mice induced by D-galactose and explore its mechanism.
      METHODS  A subacute aging model was constructed by subcutaneous injection of D-galactose into mice. Western medicine positive drug AREDS2, Chinese medicine positive drug Qiju Dihuang Pills, low and high dose groups of water extract of Jujing Pill, and low and high dose groups of 50% ethanol extract of Jujing Pill were administered, respectively. Optical coherence tomography (OCT) was used to evaluate the condition of mouse retina and fundus, HE staining was adopted to assess retinal cell damage, TUNEL staining was applied to observe retinal cell apoptosis, kits were used to measure the expression of oxidative stress factors, and western blotting was used to detect the expression of retinal apoptosis and oxidative stress pathway related proteins in mice.
      RESULTS  The extracts of Jujing Pills significantly increased the retinal thickness (P < 0.05), improved the retinal injury and inhibited the apoptosis of retinal cells in model mice (P < 0.05). At the same time, Jujing Pills down-regulated the expression of apoptotic proteins such as Cleaved Caspase-3 and Cleaved Caspase-9 (P < 0.05) while up-regulated the expression ratio of Bcl-2/Bax (P < 0.05). Jujing Pills also significantly increased the expression of antioxidant factors SOD and GSH (P < 0.05), decreased the production of oxidative product MDA (P < 0.05), and significantly up-regulated the expression of antioxidant proteins Nrf2 and HO-1(P < 0.05). The water extracts of Jujing Pills exerted similar retina protection effect with the 50% ethanol extracts.
      CONCLUSION  Jujing Pills can improve the retinal injury of subacute aging model mice, which is related to the inhibition of retinal cell apoptosis and the improvement of oxidative stress.

     

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