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[辣木黄酮对糖尿病性脑病大鼠认知功能及神经病理学指标的影响]

[Effects of Moringa flavone on cognitive function and neuropathological indexes in diabetic encephalopathy rats].

作者信息

Zhu Jianzhong, Zhao Can, Qiao Yuebing, Liu Yuanyuan, Sui Yuelin

机构信息

Department of Anatomy, Cangzhou Medical College, Cangzhou 061000, Hebei, China. Corresponding author: Sui Yuelin, Email:

出版信息

Zhonghua Wei Zhong Bing Ji Jiu Yi Xue. 2020 Dec;32(12):1491-1495. doi: 10.3760/cma.j.cn121430-20200330-00459.

DOI:10.3760/cma.j.cn121430-20200330-00459
PMID:33541503
Abstract

OBJECTIVE

To investigate the effect of Moringa flavone on cognitive impairment and neuropathological indexes in diabetic encephalopathy (DE) rats.

METHODS

Sixty male Sprague-Dawley (SD) rats were divided into control group, model group, positive drug group, Moringa low-dose and high-dose groups according to the random number table method, with 10 rats in each group. Diabetic rat model was established by intraperitoneal injection of 25 mg/kg streptozotocin (STZ) after continuous feeding of high fat and high sugar diet for one week. Blood was collected from the tail vein after 72 hours, the mean value of twice random blood glucose was ≥ 16.67 mmol/L, and the continuous positive urine glucose showed that the diabetes model was successfully prepared. The control group was fed with conventional feed. After successful model establishment, the rats in the Moringa low and high dose groups were given 4.0 g/kg and 8.0 g/kg Moringa extract (Moringa flavone) by gavage everyday, the rats in the positive drug group were given piracetam 0.48 g/kg, and the rats in the model group and control group were given the same amount of normal saline once a day for 30 days. Morris water maze was used to evaluate the cognitive impairment of the rats. The hippocampus of the rats was harvested 12 hours after the last administration, and the advanced glycation end product receptor (RAGE) and nuclear factor-κB (NF-κB) were detected by immunohistochemistry. The contents of acetylcholinesterase (AChE), advanced glycation end product (AGE) and choline acetyl transferase (ChAT) were detected by enzyme linked immunosorbent assay (ELISA).

RESULTS

Compared with the control group, the escape latency and the exploration distance in model group were extended, target quadrant stay time was shortened, the levels of AChE and AGE in brain tissue were significantly increased, and ChAT level was significantly decreased. Morris water maze experiment showed that compared with the model group, in the Moringa low and high dose groups from the 3rd day, the escape latency (s: 35.07±7.21, 33.14±5.35 vs. 43.09±9.83, both P < 0.05) and the exploration distance (m: 8.32±4.23, 8.10±4.97 vs. 13.02±3.67) were significantly shortened (both P < 0.05). The target qauadrant stay time was extended (s: 35.12±3.12, 41.53±8.37 vs. 23.15±4.89, both P < 0.01). The results of ELISA showed that compared with the model group, the levels of AChE and AGE in brain tissue of the Moringa low and high dose groups were significantly decreased [AChE (U/L): 180.22±12.03, 142.67±20.56 vs. 205.27±25.14, AGE (μg/L): 439.10±25.19, 428.27±19.14 vs. 501.28±21.53, all P < 0.05], and the levels of ChAT were significantly increased (U/L: 51.95±5.27, 53.13±5.04 vs. 37.91±5.10, both P < 0.01). There were no significant differences in AChE, AGE or ChAT between the Moringa low and high dose groups. The results of immunohistochemistry showed that the number of RAGE and NF-κB positive cells in DG area of hippocampus increased significantly, and the average gray values of RAGE and NF-κB decreased significantly. Compared with the model group, the RAGE and NF-κB positive cells in the Moringa low and high dose groups were significantly reduced, and the average gray values of RAGE and NF-κB in hippocampus were significantly increased [RAGE (gray value): 110.46±10.04, 117.76±8.64 vs. 92.19±8.76, NF-κB (gray value): 109.40±8.93, 116.59±7.26 vs. 90.74±13.27, all P < 0.05]. There were no significant differences in the expressions of RAGE or NF-κB between the Moringa low and high dose groups.

CONCLUSIONS

Moringa flavonoids could obviously improve the cognitive dysfunction and memory ability of DE model rats, improve the pathological changes of hippocampus, and have a certain protective effect on brain.

摘要

目的

探讨辣木黄酮对糖尿病性脑病(DE)大鼠认知功能障碍及神经病理学指标的影响。

方法

将60只雄性Sprague-Dawley(SD)大鼠按随机数字表法分为对照组、模型组、阳性药物组、辣木低剂量组和高剂量组,每组10只。连续高脂高糖饮食1周后,腹腔注射25 mg/kg链脲佐菌素(STZ)建立糖尿病大鼠模型。72小时后尾静脉采血,两次随机血糖平均值≥16.67 mmol/L,且尿糖持续阳性表明糖尿病模型制备成功。对照组给予常规饲料。模型建立成功后,辣木低、高剂量组大鼠每天灌胃给予4.0 g/kg和8.0 g/kg辣木提取物(辣木黄酮),阳性药物组大鼠给予吡拉西坦0.48 g/kg,模型组和对照组大鼠每天给予等量生理盐水,连续30天。采用Morris水迷宫评估大鼠的认知功能障碍。末次给药12小时后取大鼠海马,采用免疫组化法检测晚期糖基化终末产物受体(RAGE)和核因子-κB(NF-κB)。采用酶联免疫吸附测定(ELISA)法检测乙酰胆碱酯酶(AChE)、晚期糖基化终末产物(AGE)和胆碱乙酰转移酶(ChAT)的含量。

结果

与对照组相比,模型组大鼠逃避潜伏期延长,探索距离增加,目标象限停留时间缩短,脑组织中AChE和AGE水平显著升高,ChAT水平显著降低。Morris水迷宫实验显示,与模型组相比,辣木低、高剂量组从第3天起逃避潜伏期(秒:35.07±7.21,33.14±5.35比43.09±9.83,均P<0.05)和探索距离(米:8.32±4.23,8.10±4.97比13.02±3.67)显著缩短(均P<0.05)。目标象限停留时间延长(秒:35.12±3.12,41.53±8.37比23.15±4.89,均P<0.01)。ELISA结果显示,与模型组相比,辣木低、高剂量组脑组织中AChE和AGE水平显著降低[AChE(U/L):180.22±12.03,142.67±20.56比205.27±25.14,AGE(μg/L):439.10±25.19,428.27±19.14比501.28±21.53,均P<0.05],ChAT水平显著升高(U/L:51.95±5.27,53.13±5.04比37.91±5.10,均P<0.01)。辣木低、高剂量组之间AChE、AGE或ChAT水平无显著差异。免疫组化结果显示,海马齿状回(DG)区RAGE和NF-κB阳性细胞数量显著增加,RAGE和NF-κB平均灰度值显著降低。与模型组相比,辣木低、高剂量组RAGE和NF-κB阳性细胞显著减少,海马中RAGE和NF-κB平均灰度值显著升高[RAGE(灰度值):110.46±10.04,117.76±8.64比92.19±8.76,NF-κB(灰度值):109.40±8.93,116.59±7.26比90.74±13.27,均P<0.05]。辣木低、高剂量组之间RAGE或NF-κB表达无显著差异。

结论

辣木黄酮可明显改善DE模型大鼠的认知功能障碍和记忆能力,改善海马病理变化,对脑具有一定的保护作用。

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