Experiments on the effect of drugs on the hemolysin content in animals
Experiments on the effect of drugs on the hemolysin content in animals
This experiment is mainly used to test the effect of drugs on hemolysin content in animals.
Operation method
Cell Culture Technology
Principle
Effect of drugs on hemolysin content in animals The effect of drugs on hemolysin in mice was experimentally determined and statistically analyzed using SPSS software.
Materials and Instruments
Mouse Move 1. 80 mice of Kunming breed, weighing 18~22 g, were randomly divided into 8 groups, i.e., blank control group, normal Liaoning ginseng gavage group, normal Jilin ginseng gavage group, normal Korean ginseng gavage group, immunocompromised model control group, immunocompromised Liaoning ginseng treatment group, immunocompromised Jilin ginseng treatment group, and immunocompromised Korean ginseng treatment group, with 10 mice in each group, and half of each group being of both sexes. Caveat Blood should be taken and used now. For more product details, please visit Aladdin Scientific website.
Sheep erythrocytes Complement Doxa reagent Ginseng Hydrocortisone Injection
Micro-sampler Thermostatic Shaker UV-Vis Spectrophotometer
2. After keeping the animals for 3 days, the mice in the immunocompromised model group and the immunocompromised treatment group of the three local ginsengs were injected subcutaneously with hydrocortisone injection, 70 mg/kg (body weight), once/d for 1 week.
3. When the model was completed, the blank control group and the immunocompromised model group were fed normally with unlimited diet; the normal three-land ginseng gavage group and the three-land ginseng immunocompromised gavage group were gavaged with three-land ginseng decoction, 5 g/kg (body weight)/d (i.e., 100% decoction, 0.2 mL/pupil), 1 time/d for 2 consecutive weeks, respectively.
4. When the treatment was completed, blood was taken from the eyes of mice to isolate the serum.
5. Serum hemolysin content was determined by washing sheep red blood cells (SRBC) with saline 3 times (1000-2000 r/min, 5 min), discarding the supernatant, and 3:5 (V/V) diluting the SRBC to 2x109 cells/mL with saline.
6. 4 days before sampling, mice were injected intraperitoneally with 0.2 mL of prepared SRBC.
(1) On the day of sampling, mice had their eyeballs removed to obtain blood, serum was separated, and the serum was diluted 500-fold.
(2) Take 1 mL of diluted serum and add it to the reaction tube, then add 0.5 mL of SRBC, place it in an ice bath and add 1 mL of complement to each tube.
(3) Transfer the reaction tubes to a 37°C water bath for 10 minutes and then to an ice bath.
(4) Centrifuge at 2000 r/min for 10 min after the ice bath, take 1 mL of supernatant, add 3 mL of Doxa reagent and mix well, and then measure the absorbance by spectrophotometer (540 nm) after 10 min.
7. Determine the absorbance value of half hemolysis of SRBC used in the experiment, and take 0.25 mL of the same SRBC used in the experiment, add 4 mL of Doxor's reagent, shake well, and then determine the absorbance with a spectrophotometer after 10 min, which will be used as a control for half hemolysis.
8. The conversion formula is as follows: sample HC50 = (sample absorbance value x dilution) / SRBC half hemolyzed absorbance.
9. Statistical analysis The data of each group were expressed as (x ± s) and analyzed by ANOVA using SPSS17.0 statistical software.