Efficacy Evaluation of Qishen Yizhi Formula in Improving the Learning and Memory Ability of D-Galactose Induced Subacute Aging Mice
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摘要:
目的 评价杞参益智方对D-半乳糖注射亚急性衰老模型小鼠学习与记忆能力的改善效应。 方法 小鼠皮下注射D-半乳糖构建亚急性衰老动物模型, 分别给予阳性药多奈哌齐(2 mg·kg-1·d-1)、杞参益智方低剂量组(1.33 g·kg-1·d-1)、高剂量组(2.67 g·kg-1·d-1)，连续给药30 d。通过水迷宫和Y迷宫实验评价模型小鼠学习与记忆行为; 采用HE染色考察模型小鼠海马损伤情况; 采用TUNEL法检测小鼠海马组织凋亡情况; 采用ELISA法检测小鼠海马组织中氧化应激因子与炎症因子表达水平; 采用Western blot法检测小鼠海马凋亡、氧化应激与炎性应激相关信号通路蛋白表达。 结果 杞参益智方水提取物显著缩短模型小鼠水迷宫测试中潜伏期和上台前路程(P＜0.01), 增加穿越平台次数(P＜0.01);增加模型小鼠Y迷宫新臂探索时间和次数(P＜0.05);抑制模型小鼠海马TUNEL荧光表达(P＜0.01);上调氧化应激因子超氧化物歧化酶(SOD)活力(P＜0.05)与谷胱甘肽(GSH)含量(P＜0.05), 下调丙二醛(MDA)含量(P＜0.05);下调白细胞介素(IL)-1β、IL-6与肿瘤坏死因子(TNF-α)表达水平(P＜0.05，P＜0.01);下调凋亡信号通路蛋白Cleaved Caspase-3与Caspase-3表达(P＜0.05)，上调氧化应激信号通路蛋白Nrf2与HO-1表达(P＜0.05)，下调炎性应激信号通路蛋白p-NF-κB与NF-κB表达(P＜0.05)。 结论 杞参益智方具有改善D-半乳糖注射亚急性衰老模型小鼠学习与记忆能力的作用, 可能与其抑制海马氧化应激与炎性应激作用相关。 Abstract:OBJECTIVE To evaluate the effect of Qishen Yizhi formula on improving learning and memory ability in D-galactose subcutaneous injection induced subacute aging mice. METHODS Subacute aging mice model mice were developed by D-galactose subcutaneous injection and then treated with positive drug donepezil (2 mg·kg-1·d-1) and Qishen Yizhi formula water extracts in low (1.33 g·kg-1·d-1) and high dose group (2.67 g·kg-1·d-1). The learning and memory abilities of mice were evaluated using Morris water maze and Y maze tests; HE staining was used to examine hippocampal damage in model mice; TUNEL was used to detect apoptosis of mouse hippocampal tissue; ELISA was used to detect the expression levels of oxidative stress factors and inflammatory factors in the mouse hippocampus tissue; Western blot was used to detect the expression of signaling pathway proteins related to apoptosis, oxidative stress and inflammatory stress in the hippocampus of mice. RESULTS The water extract of Qishen Yizhi formula significantly shortened the latency and distance of model mice for reaching the platform in the water maze test (P < 0.01), and significantly increased the number of crossing the platform (P < 0.01); increased the exploration time and number of the Y maze new arm in model mice (P < 0.05); inhibited the TUNEL fluorescence expression in the hippocampus of model mice (P < 0.01); upregulated the activity of the oxidative stress factor superoxide dismutase (SOD) (P < 0.05) and glutathione (GSH) content (P < 0.05), and downregulated malondialdehyde (MDA) content (P < 0.05); reduced interleukin (IL)-1β, IL-6 and tumor necrosis factor (TNF-α) expression levels (P < 0.05, P < 0.01); decreased the expression of apoptosis signaling pathway proteins Cleaved Caspase-3 and Caspase-3 (P < 0.05), upregulated the expression of oxidative stress signaling pathway proteins Nrf2 and HO-1 (P < 0.05), and downregulated the expression of inflammatory stress signaling pathway proteins p-NF-κB and NF-κB (P < 0.05). CONCLUSION Qishen Yizhi formula can improve the learning and memory ability of subacute aging model mice injected with D-galactose, which may be related to its inhibitory effect on hippocampal oxidative stress and inflammatory stress. -
Key words:
- Qishen Yizhi formula /
- aging /
- oxidative stress /
- neuronal inflammation
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图 1 杞参益智方对D-半乳糖注射亚急性衰老模型小鼠水迷宫行为学的影响
注: 与空白组比较, #P＜0.05, ##P＜0.01;与模型组比较, *P＜0.05, * *P＜0.01。x±s，n=8。
Figure 1. Effect of Qishen Yizhi formula on behaviors of water maze in D-galactose injection induced subacute aging model mice
图 2 杞参益智方对D-半乳糖注射亚急性衰老模型小鼠Y迷宫行为学的影响
注: 与空白组比较, #P＜0.05, ##P＜0.01;与模型组比较, *P＜0.05, * *P＜0.01。x±s，n=8。
Figure 2. Effect of Qishen Yizhi formula on behaviors of Y maze in D-galactose injection induced subacute aging model mice
图 3 杞参益智方对D-半乳糖注射亚急性衰老小鼠海马组织凋亡的影响(标尺=100 μm)
注: 与空白组比较, ##P＜0.01;与模型组比较, * *P＜0.01。x±s，n=6。
Figure 3. Effect of Qishen Yizhi formula on apoptosis in hippocampus of D-galactose injection induced subacute aging model mice (bar=100 μm)
图 4 杞参益智方对D-半乳糖注射亚急性衰老小鼠海马组织氧化应激因子表达的影响
注: 与空白组比较, #P＜0.05;与模型组比较, *P＜0.05。x±s，n=8。
Figure 4. Effect of Qishen Yizhi formula on the expression levels of oxidative stress factors in hippocampus of D-galactose injection induced subacute aging model mice
图 5 杞参益智方对D-半乳糖注射亚急性衰老模型小鼠海马组织炎性因子表达的影响
注: 与空白组比较, #P＜0.05, ##P＜0.01;与模型组比较, *P＜0.05, * *P＜0.01。x±s，n=8。
Figure 5. Effect of Qishen Yizhi formula on the expression of inflammatory cytokines in hippocampus of D-galactose injection induced subacute aging model mice
图 6 杞参益智方对D-半乳糖注射亚急性衰老模型小鼠凋亡通路蛋白表达水平的影响
注: 与空白组比较, #P＜0.05;与模型组比较，*P＜0.05。x±s，n=5。
Figure 6. Effect of Qishen Yizhi formula on the expression of apoptotic proteins in hippocampus of D-galactose injection induced subacute aging model mice
图 7 杞参益智方对D-半乳糖注射亚急性衰老模型小鼠海马组织Nrf2和HO-1蛋白表达水平的影响
注: 与空白组比较, #P＜0.05;与模型组比较，*P＜0.05。x±s，n=5。
Figure 7. Effect of Qishen Yizhi formula on the expressions of Nrf2 and HO-1 proteins in hippocampus of D-galactose injection induced subacute aging model mice
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