Effects of quantum lipid-lowering instrument on learning and memory ability and antioxidant capacity of natural aging mice

Sun Dechun¹ Wang Chao² Gao Lei² SunJian¹ Li Zhicai³ Li Xiaowen¹ Li Shengyu²

1. Suzhou Institute of Biomedical Engineering Technology, Chinese Academy of Sciences, Suzhou 215163, Jiangsu, China;

2. Agricultural Products Processing Institute, Jilin Academy of Agricultural Sciences, Changchun 130033, Jilin, China;

3. Changchun Xianying Medical Technology Co., Ltd., Changchun, Jilin 130103

【Summary】

Objective: To observe the anti-aging effect of quantum lipid-lowering instrument on natural aging mice, and to provide experimental basis for its further application.

Method：Twenty 6-week-old female mice were used as the control group, and 80 11-month-old female mice were divided into 4 groups. The model group was fed with normal feeding, the positive group was given vitamin E by gavage, the low-intensity group and the high-intensity group were given a quantum lipid-lowering instrument, the intensity power output was 65 uw/cm2, and the treatment time in the high-intensity group increased. After the last treatment, each group was subjected to platform jumping test and water maze test; the changes in the body weight of the mice were observed, and the organ index was calculated; the antioxidant indexes and sex hormone indexes in the serum and liver tissue of the mice were determined. Results: Compared with the model group, the positive group and the high-intensity group could significantly prolong the time of mice jumping off the platform for the first time and shorten the escape latency. The differences in telomerase content were statistically significant. The high-intensity group could increase the distance and time percentage of the water maze test, and the difference was statistically significant compared with the model group.

Conclusion: Quantum lipid-lowering instrument exerts its anti-aging effect by improving the learning and memory ability of natural aging mice, inhibiting lipid peroxidation, and delaying telomere shortening.

Aging is a process in which various organs and tissues of the body undergo degenerative changes with age [1]. Modern research believes that aging is also a disease that can be treated and prevented. At present, many anti-aging or anti-aging drugs or new ingredients have been developed and studied [2-6]. Anti-aging medical devices are also in development.

The quantum energy released by the quantum lipid-lowering instrument is selectively absorbed by the human body and has a biological stimulation effect. It can use the inherent pathway of human blood lipid metabolism to correct the disorder of blood lipid metabolism and play a role in the treatment of dyslipidemia without causing damage to the body [7] ]. Due to the aging of the human body, there are different degrees of metabolic disorders, increased blood lipids, blood viscosity, and lipofuscin.

Therefore, whether the quantum lipid-lowering instrument has a certain anti-aging and anti-aging effect on the human body while effectively correcting the disorder of blood lipid metabolism in the human body is worthy of in-depth study. In this study, naturally aging mice were given the same quantum intensity for different treatment times to observe the effects of the quantum lipid-lowering instrument on the learning and memory ability and antioxidant capacity of model animals, and to accumulate data for the anti-aging effect of the quantum lipid-lowering instrument.

1 Materials and methods

1.1 Experimental animals

20 SPF Kunming female mice, 6 weeks old, weighing 18~22g; 80 SPF Kunming female mice, 11 months old, weighing 40~50g, all purchased from Changchun Yisi Laboratory Animal Technology Co., Ltd. Company, production license number: SCXK (Ji)-2011-0004.

1.2 Experimental instruments

Quantum lipid-lowering instrument (Jishi and Drug Administration (Zun) word 2013 No. 2260103), Changchun Xianying Medical Technology Co., Ltd.; platform jumping instrument (ZH-800S), Anhui Zhenghua Biological Instrument Co., Ltd.; Morris water maze (Morris) , Anhui Zhenghua Biological Instrument Co., Ltd.

1.3 Experimental method

1.3.1 Grouping and handling of experimental animals

Twenty SPF grade Kunming female mice, 6 weeks old, were adaptively fed with common feed for 1 week, and served as the control group. Distilled water.

80 SPF grade Kunming female mice, 11 months old, were adaptively fed with common feed for 1 week and then randomly divided into 4 groups, namely model group, positive group, low-intensity group, and high-intensity group, with 20 mice in each group, from From the beginning of the first week to the end of the fifth week of the experiment, the animals were fed with common feed, freely ingested, and freely drank distilled water. The positive treatment group was given vitamin E 40 mg/kg once a day from the 1st week to the 2nd week, rested for a week in the 3rd week, and continued to be given vitamin E 40mg/kg once a day from the 4th week to the 5th week. The high-intensity treatment group and the low-intensity treatment group were treated with a quantum lipid-lowering apparatus, and the intensity power output was 65uw/cm2. The low-intensity treatment group was treated once a day from the 1st week to the 2nd week, each treatment was 5 minutes; the 3rd week rested for a week; from the 4th week to the 5th week, the treatment was continued once a day, and each treatment was 5 minutes. The high-intensity treatment group was treated twice a day from the first week to the second week, each treatment was 10 minutes; the third week rested for one week; the fourth week to the fifth week continued to be treated twice a day, with each treatment for 10 minutes.

1.3.2 Platform test

After the last treatment, mice in each group were subjected to platform jumping test. The mice were placed in the reaction box to adapt for 5 minutes, and the 36V alternating current was immediately applied to record the time of the mouse jumping off the platform for the first time (i.e. the latency period) and the number of electric shocks (i.e. the number of errors) within 5 minutes, which were used as the academic performance. . After 24 hours, the mice were placed directly on the platform, the test was repeated, and the time for the mice to jump off the platform for the first time and the number of electric shocks within 5 minutes were recorded as memory scores.

Aging is a process in which various organs and tissues of the body undergo degenerative changes with age [1]. Modern research believes that aging is also a disease that can be treated and prevented. At present, many anti-aging or anti-aging drugs or new ingredients have been developed and studied [2-6]. Anti-aging medical devices are also in development.

The quantum energy released by the quantum lipid-lowering instrument is selectively absorbed by the human body and has a biological stimulation effect. It can use the inherent pathway of human blood lipid metabolism to correct the disorder of blood lipid metabolism and play a role in the treatment of dyslipidemia without causing damage to the body [7] ]. Due to the aging of the human body, there are different degrees of metabolic disorders, increased blood lipids, blood viscosity, and lipofuscin.

Therefore, whether the quantum lipid-lowering instrument has a certain anti-aging and anti-aging effect on the human body while effectively correcting the disorder of blood lipid metabolism in the human body is worthy of in-depth study. In this study, naturally aging mice were given the same quantum intensity for different treatment times to observe the effects of the quantum lipid-lowering instrument on the learning and memory ability and antioxidant capacity of model animals, and to accumulate data for the anti-aging effect of the quantum lipid-lowering instrument.

1 Materials and methods

1.1 Experimental animals

20 SPF Kunming female mice, 6 weeks old, weighing 18~22g; 80 SPF Kunming female mice, 11 months old, weighing 40~50g, all purchased from Changchun Yisi Laboratory Animal Technology Co., Ltd. Company, production license number: SCXK (Ji)-2011-0004.

1.2 Experimental instruments

Quantum lipid-lowering instrument (Jishi and Drug Administration (Zun) word 2013 No. 2260103), Changchun Xianying Medical Technology Co., Ltd.; platform jumping instrument (ZH-800S), Anhui Zhenghua Biological Instrument Co., Ltd.; Morris water maze (Morris) , Anhui Zhenghua Biological Instrument Co., Ltd.

1.3 Experimental method

1.3.1 Grouping and handling of experimental animals

Twenty SPF grade Kunming female mice, 6 weeks old, were adaptively fed with common feed for 1 week, and served as the control group. Distilled water.

80 SPF grade Kunming female mice, 11 months old, were adaptively fed with common feed for 1 week and then randomly divided into 4 groups, namely model group, positive group, low-intensity group, and high-intensity group, with 20 mice in each group, from From the beginning of the first week to the end of the fifth week of the experiment, the animals were fed with common feed, freely ingested, and freely drank distilled water. The positive treatment group was given vitamin E 40 mg/kg once a day from the 1st week to the 2nd week, rested for a week in the 3rd week, and continued to be given vitamin E 40mg/kg once a day from the 4th week to the 5th week. The high-intensity treatment group and the low-intensity treatment group were treated with a quantum lipid-lowering apparatus, and the intensity power output was 65uw/cm2. The low-intensity treatment group was treated once a day from the 1st week to the 2nd week, each treatment was 5 minutes; the 3rd week rested for a week; from the 4th week to the 5th week, the treatment was continued once a day, and each treatment was 5 minutes. The high-intensity treatment group was treated twice a day from the first week to the second week, each treatment was 10 minutes; the third week rested for one week; the fourth week to the fifth week continued to be treated twice a day, with each treatment for 10 minutes.

1.3.2 Platform test

After the last treatment, mice in each group were subjected to platform jumping test. The mice were placed in the reaction box to adapt for 5 minutes, and the 36V alternating current was immediately applied to record the time of the mouse jumping off the platform for the first time (i.e. the latency period) and the number of electric shocks (i.e. the number of errors) within 5 minutes, which were used as the academic performance. . After 24 hours, the mice were placed directly on the platform, the test was repeated, and the time for the mice to jump off the platform for the first time and the number of electric shocks within 5 minutes were recorded as memory scores.

1.3.3 Morris water maze test

After the last treatment, the mice in each group underwent the water maze test for 6 consecutive days. The first 5 days were the positioning navigation experiment, and the 6th day was the space exploration experiment. Positioning navigation experiment: training 4 times a day, each time the mice face the pool wall and put them into the water from different water entry points, record the time required for the mice to find the underwater hidden platform from entering the water (ie escape latency), average Swimming speed, activity time in the target quadrant within 90s, swimming distance and total swimming distance. Space exploration experiment: Remove the underwater platform, use a position in any quadrant of the pool as the water entry point, let the mice swim in the pool to find the platform, record the time required for the mice to pass the platform for the first time, the number of times they pass the platform, the average swimming speed, The time spent in the target quadrant within 90s, the swimming distance and the total swimming distance. The calculation formula of distance percentage and time percentage is as follows: distance percentage = swimming distance in target quadrant/total distance; time percentage = swimming time in target quadrant/total time.

1.3.4 Preparation of serum and tissue samples

After the Morris water maze test, the mice were fasted for 12 hours, weighed, and the eyeballs were removed for blood, centrifuged at 4°C and 3000 r/min for 10 min, and serum was collected and placed in a -80°C refrigerator for later use. The mice were sacrificed by cervical dislocation after blood collection. The liver, kidney, spleen, ovary and brain tissues of the mice were quickly dissected out on ice packs. The floating blood was removed with cotton swabs and weighed. Then, the mouse livers were placed in ice-cold saline. Rinse in medium to remove blood, blot dry with filter paper, prepare 1% and 10% tissue homogenates with pre-cooled normal saline, centrifuge at 4°C, 4000 r/min for 10 min, take the supernatant and store it at -80°C for later use.

1.3.5 Determination of organ index

According to the weight of the mice recorded in the method 1.3.4 and the weight of each organ of the mice after dissection, the organ index of the mice was calculated according to the following formula.

Organ index (mg/g) = wet organ weight (mg)/mouse body weight (g) According to the recorded weight of the mice in 1.3.4 and the weight of each organ after dissection of the mice, according to The following formula calculates the mouse organ index.

Organ index (mg/g) = organ wet weight (mg)/mouse body weight (g)

1.3.6 Detection indicators and detection methods in serum and organ tissues

Antioxidant indicators: Superoxide dismutase (SOD), glutathione peroxidase (GSH Px), catalase (CAT) and glutathione peroxidase (GSH Px) in mouse serum and liver tissue were detected respectively. Dialdehyde (malondialdehyde, MDA) content, and serum telomerase content.

Sex hormone indexes: The content of follicle-stimulating hormone (FSH) and luteinizing hormone (LH) in serum of mice were detected respectively.

The above indicators were determined in strict accordance with the kit instructions.

1.4 Data Statistical Processing

SPSS19.0 statistical software was used to process the data, the statistical data of each detection index were expressed as mean and standard deviation (x±SD), and P<0.05 was considered as statistically significant.

2 Results

2.1 Comparison of learning and memory ability indexes of mice in each group

As can be seen from Table 1, compared with the model group, the positive group, the low-intensity group and the high-intensity group can all prolong the time for the mice to jump off the platform for the first time and reduce the number of errors in the mice within 5 minutes. There was a statistically significant difference in the latency period of mice in the high-intensity group (P<0.05). It shows that high-intensity quantum lipid-lowering instrument treatment can improve the ability of non-spatial long-term learning and memory in natural aging mice.

It can be seen from Table 2 that in the positioning navigation test, compared with the model group, the escape latency of the mice in the positive group, low-intensity group and high-intensity group was significantly shortened, and the difference between the groups was statistically significant (P<0.05). The average swimming speed of the mice in the high-intensity group increased, and the distance and time percentages of the mice in the positive group, the low-intensity group and the high-intensity group increased significantly, and the difference between the groups was statistically significant (P<0.01).

It can be seen from Table 3 that in the spatial search test, compared with the model group, the distance and time percentages of the mice in the low-intensity group and the high-intensity group were increased, and the distance and time percentages of the mice in the high-intensity group were significantly increased. higher than the model group, and the difference between groups was statistically significant (P<0.01). It can be seen that the quantum lipid-lowering instrument has improved the spatial learning and memory ability of naturally aging mice.

It can be seen from Table 4 that compared with the model group, although the organ indexes of the mice in the positive group, the low-intensity group and the high-intensity group all increased, the difference was not statistically significant (P>0.05).

As can be seen from Table 5 and Table 6, compared with the model group, the serum and liver SOD, GSH-Px and CAT contents of the mice in the positive group, low-intensity group and high-intensity group all increased, and the MDA content decreased, and the high The difference between the intensity group and the positive group and the model group was statistically significant (P<0.05).

2.3 Comparison of serum telomerase content and estrogen in mice in each group

As can be seen from Figure 1, compared with the model group, the serum telomerase concentration in the positive group, the low-intensity group and the high-intensity group of natural aging mice increased to varying degrees, and the difference was statistically significant with the model group (P <0.05). The serum telomerase concentration in the high-intensity group was higher than that in the low-intensity group, close to the positive group, indicating that the quantum lipid-lowering instrument can increase the content of telomerase in naturally aging mice and prolong the cell lifespan of naturally aging mice.

It can be seen from Table 7 that there was no significant difference in follicle stimulating hormone (FSH) and luteinizing growth hormone (LH) between the experimental groups and the control group (P>0.05).

3 Discussion

In this experiment, natural aging mice were used to establish a model, in order to maximize the closeness to the internal environment of the aging body. The significant manifestations of brain aging are the decline of learning ability and the impairment of memory. A large number of studies have confirmed that the learning and memory ability of humans and animals has an aging decline phenomenon [10-12], especially the spatial reference memory ability increases with age. declining trend [13]. Therefore, the platform jump test and Morris water maze test were used to detect the long-term learning and memory ability of mice. The results of the platform jump test showed that the incubation period of the mice in the model group was significantly shortened, and the incubation period was significantly prolonged after treatment with the quantum lipid-lowering instrument, and the number of errors within 5 minutes was reduced; Morris water maze mice had a certain degree of learning and safety in finding a safe platform during swimming training. Memory ability, the test results show that the escape latency of mice treated with the quantum lipid-lowering instrument is significantly shortened, indicating that the quantum lipid-lowering instrument has a certain therapeutic effect on naturally aging mice with severely impaired learning and memory abilities.

Oxidative damage is the mainstream theory of cell aging, and free radicals can easily damage unsaturated fatty acids in cell membranes, resulting in lipid peroxidation [14]. SOD is an important substance for scavenging free radicals in the body. In anti-aging research, the detection of SOD activity can be used as an important indicator to judge the degree of aging[15-16], and its activity can reflect the body's ability to scavenge free radicals[17]; GSH-Px It can catalyze the reductive reaction with H2O2 to generate harmless water and oxidized GSH, which is beneficial to protect the integrity of cell membrane structure and function[18]; CAT is ubiquitous in biological organisms and protects the body from harmful substances [19] ]; MDA is the product of lipid peroxidation, which can indirectly react

The degree of oxidative damage to the body [20]. This study shows that the activities of antioxidant enzymes such as SOD, GSH-Px and CAT in mice in the high-intensity treatment group were significantly improved, and lipid peroxidation products such as MDA were reduced, suggesting that quantum lipid-lowering instrument can improve the scavenging of free radicals in naturally aging mice. ability, reduce the damage of peroxidation products to the body, inhibit lipid

Peroxidation, thereby reducing the aging degree of the body and exerting its anti-aging effect.

Telomere is a special structure that exists at the end of eukaryotic chromosomes. Its length is shortened as the cell divides. When it shortens to a certain extent, the cell stops dividing and tends to age and die. Length determines the lifespan of cells

life [21]. Telomerase is a nuclear protein reverse transcriptase, and its active expression can stabilize telomere length to prevent it from being shortened during replication [22-23], and cell lifespan can be prolonged. This study shows that the quantum lipid-lowering instrument can increase the activity of telomerase in the serum of naturally aging mice, and delaying the shortening of telomeres may be one of the mechanisms of its anti-aging effect, but the activation of telomerase is affected by multiple factors[24- 25], so the regulation mechanism of quantum lipid-lowering instrument on its activity needs to be further studied.

The degree of aging of the body is also reflected in the changes in the index of organs such as the brain, thymus, spleen, liver, and kidney [26]. However, the results of this study suggest that the quantum lipid-lowering instrument can increase the organ index of naturally aging mice, but it has no significant effect, indicating that within a certain quantum output power range (65uw/cm2), it will not significantly affect the organ quality. affected.

In addition, since female mice were used as the research objects, the content of follicle-stimulating hormone (FSH) and luteinizing growth hormone (LH) in the serum of mice was measured and studied. Sex hormones have an effect, indicating that the quantum lipid-lowering instrument is safe.

In summary, the quantum lipid-lowering instrument can improve the learning and memory ability of naturally aging mice, as well as the activities of SOD, GSH-Px and CAT antioxidant enzymes in serum and liver, reduce MDA lipid peroxidation products, and inhibit lipid peroxidation. It can reduce the aging degree of the body; and can increase the telomerase activity in the serum of naturally aging mice, delay the shortening of telomeres, and exert its anti-aging effect, but it will not affect the organs and estrogen of naturally aging mice.