Study On The Nootropic Effect Of Total Glycosides Of Cistanche And Lycium Barbarum Polysaccharides On C57BL/6J Mice Of Different Ages

Summary
Objective: To investigate the effects of the compatibility of total glycosides of Cistanche deserticola and Lycium barbarum polysaccharides on the learning and memory ability, the synthesis and degradation of cholinergic neurotransmitters in the brain, and the level of oxidative stress in C57BL/ 6J mice of different months.

Oringin of Cistanche Deserticola 

Methods: 48 C57BL/6J mice aged 3, 6, and 9 months were randomly divided into control group, total glucosides of Cistanche 100 mg/kg group, Lycium barbarum polysaccharides 100 mg/ kg group, combined (total glycosides of Cistanche 100
mg/ kg+ Lycium barbarum polysaccharide 100 mg/ kg) group.
After intragastric administration for 30 days, the learning and memory abilities of the mice in different age groups were evaluated by Morris water maze test, dark avoidance test, and platform jumping test; the ACh E, Ch AT, T-SOD, GSH-Px, MDA levels.
Results: Compared with the control group, the learning and memory abilities of the mice in the combination groups of all months were improved, the activity of ChAT and T-SOD in the cortex of the mice in the combination groups of 3 and 6 months were significantly increased, and the mice in the combination group of 9 months cortex
The activities of ChAT and GSH-Px were significantly increased, and MDA in the cerebral cortex of mice in the 6- and 9-month-old combined group
content decreased significantly (P<0. 05 or P<0. 01).

Conclusion: Compatible administration of total glycosides of Cistanche deserticola and Lycium barbarum polysaccharides can improve 3, 6, 9
The learning and memory ability of month-old C57BL/6J mice may be related to the regulation of the synthesis and degradation of cholinergic neurotransmitters in the brain, and the inhibition of oxidative stress in the brain.
Keywords: total glycosides of Cistanche deserticola; Lycium barbarum polysaccharide; Acetylcholine; Oxidative stress

1Materials and Instruments
1.1 Experimental animals
48 C57BL/6J mice of SPF grade 3, 6, and 9 months old, half male and half male, were purchased from Nanjing Jun Bioengineering Co., Ltd., experimental animal certificate number: 20180006018886.
During the experiment, free to eat and drink, 12 hours of alternating light and dark lighting, all experiments were carried out according to the experimental regulations,
The operation and management complied with the relevant regulations on the management of experimental animals.
1.2 Drugs and reagents
Total glycosides of Cistanche deserticola (mass fraction ≥80%, batch number: JZ20092513), Lycium barbarum polysaccharide (mass fraction ≥90%, batch number: JZ21042710), Nanjing Jingzhu Biotechnology Co., Ltd.; BCA kit, acetylcholinesterase (ACh E) kit , acetylcholine transferase (Ch AT) kit, malondialdehyde (MDA) kit, total superoxide dismutase (T-SOD) kit, glutathione catalase (GSH-Px) kit, Nanjing Jiancheng Institute of Bioengineering.

1. 3Main instruments

DMS-2Morris water maze experimental device, DTT-2 mouse platform jumping device, SBA-2 mouse dark avoidance test device, Institute of Materia Medica, Chinese Academy of Medical Sciences; 4503R low-temperature high-speed centrifuge
, Eppendorf, Germany; VARIOSKAN FLASH microplate reader, American Thermo Company.

2 methods
2.1 Animal grouping and administration
48 C57BL/6J mice aged 3, 6, and 9 months were randomly divided into 4 groups, and each group
12 rats, half male and half female, including blank control group, total glucosides of Cistanche deserticola (100 mg/kg), polysaccharides from Lycium barbarum (100 mg/kg), combined group (100 mg/kg total glycosides of Cistanche deserticola + 100 mg/kg wolfberry polysaccharides
mg/kg), administered by intragastric administration daily for 1 month continuously.

2. 2Morris
water maze experiment
The device consists of a black circular pool, a transparent circular platform, a camera and an automatic image acquisition and processing system. On the outer edge of the pool, it is divided into four water entry points: east, west, south and north.
, A circular transparent platform with a diameter of 15 cm is placed in the pool.
During the experiment, maintain a constant temperature, sound insulation, and no direct light to prevent external factors from interfering with the mice.
This experiment was carried out for 6 days in total: the first day to the fifth day was the positioning and navigation test, and the time for the mouse to find the platform within 120 s was recorded as the escape latency. If the mouse did not find the platform within 120 s, the escape latency was 120 s. After the training, no matter whether the mice found the platform or not, they put them on the platform to study for 15 seconds, and entered the water twice a day from the adjacent corresponding objects of the platform; The time of crossing the platform for the first time and the number of times of crossing the platform within 120s within the corresponding limit.

2.3 Darkness avoidance experiment
The device is divided into two chambers, light and dark, and the two chambers are connected by a small hole for mice to enter and exit. The bottom of the dark chamber is connected with 40 V current, and the bottom of the light chamber is not energized. The experiment was divided into two phases: the first day was the learning phase, the mice were put into the bright room with their backs facing the hole, and allowed to move freely in the two rooms for 2 to 3 minutes to adapt to the environment, then put the mice back into the light room with the hole , the habit of the mice tending to the dark and avoiding the light, and the habit of drilling holes will cause them to enter the dark room and receive electric shocks. If the mice do not enter, they will be driven to enter and make them produce memories; the second
The first day was the test period, the mice were put into the light room with their backs to the hole, and the latency period and the number of times the mice entered the dark room from the lightroom within 3 min were recorded as the test scores.