Team:UST Beijing/AnimalExperiment
Animal Experiment
We divided 20 male hamsters into 4 groups, and fed them with different diets, to test the impacts of notoginseng on cholesterol in blood. It turned out that notoginseng has positive effects on suppressing blood lipid level increase under high-fat/cholesterol diet. However, the gap of experimental data between each hamster in each group is greater than the gap between each group. One possible reason is that animals were mixed up during behavior tests.
Mixed feed
The aim of this experiment is to measure how is the effect of notoginseng on the behavior and blood lipid content of hamsters.
Experiment subject: 20 male hamsters (8 weeks old)
Condition: lighting 12h, dark 12h
Feed formulation:
*Acid treating: soak notoginseng root in 30% acetic acid and steam in order to destroying the sugar bond of saponins, after that, dry down root until it become odorless.
Experiment data: Dietary intake (individual average of each group):
Weight (g) :
We can see that ones fed by high-fat feeding stuff without notoginseng grew more rapidly than others. But the feeding process isn’t long enough, the differences between groups are small.
Mirror test
The mirror test, sometimes called the mark test or the mirror self-recognition test (MSR), is a behavioural technique as an attempt to determine whether a non-human animal possesses the ability of self-recognition. Animals are marked and given access to a mirror. If the animal then touches or investigates the mark, it is taken as an indication that the animal perceives the reflected image as itself, rather than of another animal. We wonder whether notoginseng has an effect on self-recognition of hamsters.
We built a special room for this experiment, one of the walls of which was made by a big mirror. This room was separated into two, which were totally isolated. We put group blank and group experiment in these two room, and let them go around freely. We used camera to record the hamsters’ behavior.
Finally, we found that both two group are insensitive to the mirror, which meant that notoginseng may do not have much effect on their self-recognition.
Ball-rolling experiment
We bought some balls which the hamsters can push them and make themselves go ahead. We wanted to judge the energy of the hamsters through distance of rolling balls. So we would know how the effect of notoginseng on the intelligence of hamsters. We put hamsters of different groups into balls with different color.
We chosed a wonderful morning to do experiment. We placed balls on a background and let them move freely. We observed their behavior and measured the distance of movement tracks.
We found that groups fed by notoginseng were more active than other groups.
Strength test
As we wanted to know the developmental state of hamsters’ muscle, we designed an interesting experiment. We chosed a basket made of wire and let hamsters grasp it, then we started to flip the basket until hamsters fell from it. We judge strength abilities through the time of hamsters holding on the basket.
The result is as graphs shows.
Groups fed by notoginseng held longer than group blank.
Creep Distance test
We also wanted to know hamsters’ abilities of discovery. We put a group of hamsters in center of board, and let them move freely. After 10 minutes, we measured the distance of creep.
The result is as graphs shows.
From data, hamsters in the blank group moved the shortest distance and experiment group shew a better movement ability.
Cholesterol Measurement
We dissected hamsters and took their blood after anaesthesia (almost 3mL from each hamster). Hamsters livers were removed and on cryopreservation at -80℃. Let the blood stand for thirty minutes before Centrifugation for two minutes (12000r/min). The supernatant was put into 1.5ml centrifuge tube with mark. Add sodium azide to prevent corruption, and then we stored the solution in refrigerator at 4℃.
Cholesterol separation with FPLC: FPLC is Fast protein liquid chromatography which is specially used for separation of proteins, polypeptides and polynucleotides. FPLC is an important innovation in recent years because It not only maintains the characteristics of HPLC (fast, high resolution and so on), but also has the characteristics of large capacity, high recovery efficiency and causing less loss of biological macromolecules. In recent years, it has been widely used in the separation of proteins, peptides and oligonucleotides. Parameter setting: mobile phase: PBS; Current Speed: 0.4ml/min; Collecting pipe: 45; Each tube: 0.5ml; Molecular sieve chromatography column According to the different density of lipoprotein, we separate them based on the different speed of movement in the molecular sieve. We can tell when and what kind of lipoprotein is collected under the detection of UV280nm. The collection tube is sequentially numbered from 1~45 before storing them in refrigerator at 4℃.
Cholesterol and triglyceride detection: We bought special kits to detect cholesterol and triglycerides. Principle of cholesterol detection:
Principle of triglyceride detection:
We use the 96-well-plate to detect cholesterol and triglyceride concentration of 45 samples collected from one hamster at the same time. The 48 holes on the left were for cholesterol detection, and the right side of the 48 holes were for triglyceride testing. Observe the results of Cholesterol and glycerin three fat form 45 samples: there were15 mice having lower cholesterol levels. But only five hamsters at high cholesterol level were from each of the four experimental group.
