Effects Of Total Glycosides Of Cistanche On Learning And Memory Impairment in Mice

Gao Cong 1, Wang Chuanshe 2, Wu Guanzhong 1, Tu Pengfei 2 (1. Pharmacology Teaching and Research Office of China Pharmaceutical University, Nanjing 210009; 2. Medical Department of Peking University, Beijing 100083)

Abstract: Objective To observe the effect of total glycosides of Cistanche deserticola on scopolamine-induced learning and memory acquisition disorder and sodium nitrite-induced learning and memory consolidation disorder in mice. Methods The learning and memory functions of mice were observed by the step-down method. After continuous intragastric administration of total glycosides of Cistanche deserticola for 30 days, the animals were trained on the 29th day of the experiment and tested on the 30th day. The latency of the first step and the number of errors within 5 minutes were recorded. The scopolamine model was injected intraperitoneally 15 minutes before training, while the sodium nitrite model was injected subcutaneously immediately after training. Results Compared with the scopolamine model group, the incubation period of total glycosides of cistanche 400, 200 mg/kg group and Naofukang group were significantly prolonged, and the number of errors was significantly reduced; The incubation period of the total glycosides of Cistanche deserticola 100mg/kg group was also significantly prolonged, but the number of errors was only reduced to a certain extent. Compared with the sodium nitrite model group, the incubation period of the total glycosides of cistanche 400 and 200 mg/kg groups was significantly prolonged, and the number of errors decreased to a certain extent, but there was no statistical significance; In the total glycosides of cistanche 100mg/kg group, the latency and the number of errors were only improved to a certain extent; The latency of Naofukang group was significantly prolonged, and the number of errors was significantly reduced. Conclusion The total glycosides of Cistanche can significantly improve the learning and memory acquisition disorder caused by scopolamine and the learning and memory consolidation disorder caused by sodium nitrite in mice.

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Key words: total glycosides of cistanche deserticola; Mice; Learning and memory disorders; Jumping platform method

Chinese Library Classification No.: R285.5    Document ID: A   Article  No.: 1003-9783 (2005) 03-0162-03

The aging population has searched for safe and effective drugs for the prevention and treatment of Alzheimer's disease a hot spot in drug research. In recent years, Chinese medicine has made great progress in the treatment of Alzheimer's disease with drugs that mainly invigorate the kidney and marrow and also invigorate the heart and spleen. Cistanche deserticola ma, a traditional Chinese medicine, was originally published in Shennong's Classic of Materia Medica and has the effects of tonifying kidney yang, benefiting blood essence, moistening intestines, and relieving constipation. It is now widely used to treat senile dementia, impotence, habitual constipation, and other diseases [1,2]; Studies have shown that it has the effects of anti-dementia, anti-oxidation, anti-hypoxia, anti-radiation and enhancing immune function; The main component is phenylethanolamine glycosides [3-8]. The total glycosides of Cistanche deserticola are extracted from the dried fleshy stems of Cistanche deserticola (Schenk) bright, containing 90.7% of phenylethanoid glycosides and 44.3% of the main component echinacoside. In this paper, the effect of total glycosides of cistanche deserticola on the learning and memory impairment of mice caused by scopolamine and sodium nitrite was studied to provide evidence for its development as a new drug for promoting intelligence and preventing and treating Alzheimer's disease.

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1 .Material and methods

1.1 Animals

180 healthy Kunming mice, weighing 18~22g, half male and half female, were provided by the Animal Center of China Pharmaceutical University, with the certificate number Su Dong (Zhi) 97004.

1.2 Drugs and reagents

Total glycosides of cistanche deserticola, provided by the Chemistry Department of the Modern Research Center of Traditional Chinese Medicine, Peking University, with batch number 980601. Naofukang tablets, produced by Yixing Pharmaceutical Factory, batch number 980521. Scopolamine injection, produced by Shanghai Hefeng Pharmaceutical Co., Ltd., batch number 980101. Sodium nitrite, produced by Rugao Huarui Reagent Factory, batch number 980220. The above drugs and reagents are prepared with distilled water to the required concentration for the experiment before use.

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1.3 Instrument

The wx-Z mouse platform jumping instrument is produced by the Institute of Medicine, Chinese Academy of Medical Sciences.

1.4 Method

1.4.1 Replication and test of scopolamine-induced memory acquisition disorder model in mice [9]

According to body weight, mice were randomly divided into normal group, model group, Naofukang group, and high, medium, and low dose groups of total glycosides of cistanche, with 15 mice in each group. The normal group and model group were given the same volume of distilled water by gavage once a day for 30 days.

The platform diving training was conducted on the 29th day of the experiment. The drug was administered by gavage 1 hour before the training, and scopolamine 1 mg/kg was injected intraperitoneally in each group except the normal control group 15 minutes before the training. During training, put the mice into the reaction box of the platform tester to adapt to the environment for 3min, and then immediately apply 36V alternating current. After receiving the electric shock, most of the mice jumped onto the platform to escape the electric shock. After jumping off the platform and receiving the electric shock, the mice jumped back to the platform for 5min. After 24h, put the mouse on the platform and record the time (latency) of the first time the mouse jumped off the platform and the number of electric shocks (error number) within 5min.

1.4.2 Copy and test of memory consolidation disorder model in mice caused by sodium nitrite [9]

The grouping, administration, training, and test were the same as before, but scopolamine was not injected intraperitoneally 15 minutes before training, but 120mg/kg sodium nitrite was injected subcutaneously immediately after training.

1.5 Statistical processing

The results of latency and the number of errors within 5 min are expressed in "x ± s", and the data are statistically processed by the inter-group t-test.

2.Results

2.1 Effect on scopolamine-induced learning and memory impairment in mice

Compared with the normal group, the latency of the step-down test in the model group was significantly shortened and the number of errors was significantly increased, indicating that the learning and memory acquisition of the mice was impaired. Compared with the model group, the latency of the test in the Naofukang group and the high-dose and medium-dose groups of total glycosides of cistanche were significantly prolonged, and the number of errors was significantly reduced (P<0.01); The test latency of mice in the low dose group of total glycosides of cistanche was also significantly prolonged (P<0.01); The number of test errors decreased to a certain extent, but there was no significant difference compared with the model group. The results showed that the total glycosides of cistanche could improve the learning and memory impairment of mice induced by scopolamine, and the action intensity of the high and middle-dose groups was better than that of the Naofukang group. The results are shown in Table 1.

Table 1   Effect of total glycosides of cistanche on scopolamine-induced learning and memory impairment in mice (x ± s)

Note: Compared with the model group, * P<0.05, * * P<0.01. () is the number of deaths from electric shock, the same below.

2.2 Effect on learning and memory consolidation disorder of mice caused by sodium nitrite

The latency of the step-down test of the model group mice was shortened, and the number of test errors was increased, which was significantly different from that of the normal group, indicating that the model group mice had difficulties in learning and memory consolidation. Compared with the model group, the test latency of mice in the high and middle dose groups of total glycosides of cistanche was significantly prolonged (P<0.05~0.01); The number of errors decreased to a certain extent, but there was no statistical significance; The latency and the number of errors in the test of mice in the low dose group of total glycosides of cistanche also improved to some extent, but there was no statistical significance; In Naofukang group, the latency of test was significantly prolonged and the number of test errors was significantly reduced (P<0.01). The results of this experiment show that the oral administration of high and medium doses of total glycosides of cistanche can significantly improve the learning and memory consolidation disorder of mice caused by sodium nitrite, but it is weaker than Naofukang. The results are shown in Table 2.

Table 2   Effect of total glycosides of cistanche on learning and memory consolidation disorder in mice induced by sodium nitrite (x ± s)

Note: Compared with the model group, * P<0.05, * * P<0.01.

3. Discussion

Learning and memory are the advanced functions of the brain and the elements of intelligence.

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The whole neural activity of learning and memory can be divided into three basic neural processes: acquisition, consolidation, and reproduction. Learning and memory disorders are common symptoms of various brain diseases, including cerebral vascular sclerosis, cerebral blood insufficiency, and dementia in the elderly.

Traditional Chinese medicine believes that Alzheimer's disease is mostly a deficiency syndrome. Deficiency of kidney essence and deficiency of heart and spleen lead to a void of marrow sea, which leads to a stupor. "Desert ginseng" Cistanche deserticola is a traditional tonic and aphrodisiac, with the functions of tonifying the kidney, filling the essence, and moistening the intestines. The Classic of Materia Medica calls it the essential medicine for tonifying the blood essence and nourishing the kidney. It has a good effect on the symptoms of fatigue, forgetfulness, hearing loss, and enuresis caused by kidney deficiency. Zhu Zhiming and others analyzed the drug composition of 46 compounds for the treatment of senile dementia published in relevant magazines from 1990 to 1999 and found that Cistanche deserticola is one of the most commonly used drugs [1]. 

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The animal experiment of Sato Chow in Japan confirmed that Cistanche deserticola has an anti-dementia effect [3]. Our previous experiments showed that the total glycosides of Cistanche deserticola significantly promoted the learning and memory function of normal mice, and significantly protected the learning and memory function of ethanol-induced learning and memory reproduction disorder model mice, kidney-yang deficiency model mice, vascular dementia model rats, etc. Combined with the results of this experiment, it is confirmed that Cistanche deserticola has the effect of promoting intelligence, and has obvious protective effects on the acquisition, consolidation, and reproduction of three basic neural activities in the whole neural activity of learning and memory, which shows that the effect of Cistanche deserticola glycosides on promoting intelligence is multi-level and multi-link. Therefore, total glycosides of cistanche may become an effective drug for the treatment of Alzheimer's disease and other diseases with learning and memory disorders as the main symptoms.

reference:

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[2] Yu Fang. Progress in clinical application of Cistanche deserticola [J]. Inner Mongolia Medical Journal ，2003，35(6)：535-536.

[3]satoT，Kozmas，KobayashiK，etal.Pharmaco logical studieson Cis-tanch isHerba.I.Effect softheConstituents ofCistanchis Herbaon sexandLearning Behaviorin Chronic stressed Mice(1)[J].YakugakuZasshi，1985，105(12)：1131-1144.

[4] Lei Li, Song Zhihong, Tu Pengfei. Research progress on chemical constituents of Cistanche [J]. Chinese Herbal Medicine, 2003, 34 (5): 473-476

[5] Liang Minghua, Sun Peiyi, Cao Yi. Effect of total glycosides of cistanche on lipid peroxidation during hemorrhagic shock/reperfusion injury in rabbits [J]. China Microcirculation, 2000, 4 (4): 230-231

[6] Shang Xiaoying, Wang Xiaowen, Wang Xuefei, et al. Protective effect of total glycosides of cistanche on immune function of mice injured by 60Coy radiation [J]. Chinese Herbal Medicine, 2001, 32 (2): 139-142

[7] Wu Liya Yiming, Wang Xiaowen, Zaomi Nyazi. Protective effect of total glycosides of cistanche on cerebral hypoxia in mice [J]. Journal of Xinjiang Medical University, 2003, 26 (6): 561-562

[8]MuteliefuGulinuer，LiuMingju，LuJingfen.EffectofCistanosideCom-poundson0xidativestressandImmunity[J].JournalofChinesePhar-maceuticalsciences，2001，10(3)：157-160.

[9] Zhang Juntian. Modern pharmacological experimental methodology [M]. Beijing: Beijing Medical University, China Union Medical University Joint Press, 1998.102-1021029-1031