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

2.3 Darkness avoidance experiment for mice taking cistanche

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 40V current, and the bottom of the light chamber is not energized.
The experiment was divided into two phases:
The first day is the learning stage, the mice are placed in the bright room with their backs to the hole, and let them move freely in the two rooms
After adapting to the environment for 2 to 3 minutes, put the mouse back into the hole and put it into the light room. The habit of the mouse to avoid the light and preferring to drill holes will cause it to enter the dark room and receive an electric shock. If the mouse does not enter, then drive it to enter. Make it produce memory; the second day is the test phase, put the mice back to the hole in the light room, record the latency period and the number of times the mouse enters the dark room from the lightroom within 3 minutes as the test score.

Cistanche total glycosides Function Details

2.4 Platform experiment for mice taking cistanche

The platform device is a 20cm×20cm×30cm cuboid, and an insulating rubber platform for mice to stand on is placed on the bottom surface and 36V current is passed through. The experiment was carried out for 2 consecutive days, and the first day was the learning and training stage. First, the mice were placed in the platform test box to move freely for 3 minutes to adapt to the environment.
Place the mice on an insulating platform, and energize the bottom copper grid for continuous stimulation for 180s. After receiving the electric shock, the mouse jumps onto the platform to avoid the shock. If the mouse does not jump onto the platform, the experimental operator manually guides it to the insulating platform;
The second day was the test phase. The mouse was placed on the insulating platform, and then the bottom copper grid was energized. The time when the mouse jumped off the platform for the first time was recorded as the latency period, and the number of times the mouse jumped off the platform within 180 s was recorded as the number of errors. If the mice did not jump off the platform in 180s, the latency was recorded as 180s.

2.5 Index detection
At the end of the behavioral experiment, the mice were decapitated, and the cortex was quickly separated at low temperatures, and a certain amount of cortical tissue was accurately weighed, and 9 times the volume of normal saline was added, and the homogenate was lysed into a 10% tissue homogenate.
After 10 minutes, the supernatant was taken to measure the protein concentration with the BCA kit, and the experiment was carried out strictly according to the instructions of the AChE, ChAT, MDA, T-SOD, and GSH-Px kits.

2.6 Statistical processing
All data were statistically analyzed using SPSS 19.0 software. Two-way ANOVA was used for positioning and navigation experiment data in the Morris water maze experiment, and one-way ANOVA was used for other experimental data, combined with LSD test to compare the differences among the groups.
The results are expressed in ( .x±s), and P<0.05 was considered statistically significant, and Origin2021 was used for drawing.

3 Results of the effect of cistanche

3.1 Compatibility of total glycosides of Cistanche deserticola and Lycium barbarum polysaccharide on C57BL/ 6J mice of different months
The results of the Morris water maze test are shown in Figure 1. Compared with the control group, there was no significant change in the escape latency of the 3- and 6-month-old mice in each administration group, but the escape latency of the 9-month-old mice in each administration group was significantly different on the fourth day. shortened (P<0.05). In the space exploration experiment, compared with the control group, the combined group of 6- and 9-month-old mice significantly shortened the time to cross the platform for the first time, and the number of times the mice crossed the platform increased significantly, and the time spent in the target quadrant was significantly prolonged in the combination group of mice aged 3 months. , 9-month-old Cistanche total glycosides group, the target quadrant residence time of mice was significantly prolonged (P<0. 05 or P<0. 01).

3.2 The effect of the combination of total glycosides of Cistanche deserticola and Lycium barbarum polysaccharides on the dark avoidance experiment of C57BL/ 6J mice of different ages is shown in Figure 2. Compared with the control group, the incubation period of the mice in the 3-month-old combined group was significantly prolonged, and each month-old The number of mistakes in mice in the combination group was significantly reduced, and the number of mistakes in the 9-month-old Lycium barbarum polysaccharide group was significantly reduced (P<0. 05 or P<0. 01).

3.3 Compatibility of total glycosides of Cistanche deserticola and Lycium barbarum polysaccharides at different ages
The results of the platform jumping test of C57BL/6J mice are shown in Figure 3. There was no statistically significant difference in the latent period and the number of mistakes in the platform jumping test of mice in each group.

3.4 The effect of the combination of total glycosides of Cistanche deserticola and Lycium barbarum polysaccharides on the synthesis and degradation of ACh in the cerebral cortex of C57BL/ 6J mice at different ages is shown in Figure 4. AT activity was significantly increased (P<0.05 or P<0.01).

3.5 The effects of the combination of total glycosides of Cistanche deserticola and Lycium barbarum polysaccharides on the levels of MDA, T-SOD, and GSH-Px in the cerebral cortex of C57BL/ 6J mice at different ages are shown in Figure 5. Compared with the control group, the levels of 6 and 9 months MDA in the cerebral cortex of the mice in the combination group and the mice in the 6-month-old Cistanche total glycosides group
The levels of T-SOD in the cerebral cortex of mice at 3 and 6 months were significantly increased, and the levels of GSH-Px in the 9-month-old Lycium barbarum polysaccharide group and the combination group were significantly increased (P<0.05 or P<0.01 ).

4 discussions of  the effect of cistanche

Learning and memory are one of the important advanced neurophysiological activities of the brain. Learning refers to the process of receiving external environmental information and affecting its own behavior, while memory is the process of storing, processing and extracting the acquired information or experience in the brain. It is one of the important functions for human beings to understand and adapt to the world.
Therefore, it will be a major challenge in the field of life sciences to develop drugs that improve human learning and memory functions and prevent and treat related diseases. At present, there are many behavioral testing methods for evaluating learning and memory functions, such as Morris water maze test, Y-word maze test, new object recognition test, and platform jumping test.
, Avoiding the dark experiment, etc. Among them, the Morris water maze test is a classic test for detecting the spatial learning and memory ability of rodents, and the dark avoidance test and platform jumping test are two commonly used behavioral experiments to evaluate the short-term learning and memory ability of mice.

this experiment
Using the above three behavioral experiments to evaluate the compatibility of Cistanche total glycosides and Lycium barbarum polysaccharides 3, 6, 9
Effects on learning and memory ability of month-old C57BL/6J mice. The results showed that in the Morris water maze test, the combined use of total glycosides of Cistanche deserticola and Lycium barbarum polysaccharides can significantly shorten the age of 6 and 9 months.
The time of C57BL/ 6J mice crossing the platform for the first time, increase 3, 6, 9 months old C57BL/ 6J
The number of times the mice crossed the platform and the dwell time in the target quadrant significantly prolonged the latency of the 3-month-old 57BL/6J mice in the dark avoidance test, and decreased the number of errors in the 3-, 6-, and 9-month-old C57BL/ 6J mice.
In platform jumping test, the latent period of mice in the group administered with total glucosides of Cistanche deserticola and Lycium barbarum polysaccharide in each month increased the most
, The number of errors is reduced the most. The above behavioral results suggest that compared with total glycosides of Cistanche deserticola and Lycium barbarum polysaccharides used alone, the combined use of total glycosides of Cistanche deserticola and Lycium barbarum polysaccharides can better improve the 3, 6, 9 Learning and memory ability of month-old C57BL/6J mice.