Target protein of Yimusake tablet acting on abnormal balham syndrome and erectile dysfunction syndrome models and screening method thereof
A screening method and protein technology, applied in the biological field, can solve problems such as unreported targets, lack of theory and application guidance, and unclear mechanism of action
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Embodiment 1
[0113] 1. Modeling and grouping
[0114] 25 sexually mature female mice were selected and raised under normal conditions after castration, and were used for mating experiments. Male rats were mated with female rats in estrus and combined with APO erection test to confirm that they had normal sexual function (those without normal sexual function were eliminated), and 50 rats were taken to enter the experiment, and 10 rats were randomly selected from them as normal The control group was raised in a normal environment with a temperature of 18-22°C, and the relative humidity of the environment was 40-60%. Modeling was carried out in a climate box, set temperature: 8-12°C, humidity 70-80%, fed alternately day and night for 12 / 12 hours, and placed in the same feeding environment as the normal group for other times. When the rats in the model group have dull skin and hair, lack of luster; tiredness, lying down and less movement; sensitive to stimulation, like to get together, lethar...
Embodiment 2
[0150] ELISA Detection of Drug Target Proteins in Yimusak Tablets
[0151]1) ELISA detection of the serum candidate protein of Yimusake Tablets in the rat model of abnormal mucus syndrome
[0152] Randomly select 5 kinds of serum candidate differential proteins, and perform ELISA detection on the rat model serum of the normal control group, syndrome group, and syndrome drug group, to verify whether the serum candidate differential proteins of the present invention can be used as Yimusake Tablets to challenge abnormal mucus. Drug target markers in rats:
[0153] That is, randomly select and detect No. 1 protein P01015 (angiotensinogen); No. 3 protein P02764 (α-1-acid glycoprotein); No. 4 protein P02767 (transthyretin); protein binding protein) and protein No. 14 P26644 (β2 glycoprotein 1). Double-antibody sandwich ELISA kits were used respectively (rat angiotensinogen (AGT) double-antibody sandwich kit: Wuhan USS Co., Ltd.; rat α1 acid glycoprotein (α1AG) double-antibody sand...
Embodiment 3
[0169] Immunohistochemical Detection of Drug Target Proteins in Yimusake Tablets
[0170] Randomly select 4 kinds of candidate differential proteins in penile smooth muscle tissue, and perform immunohistochemical detection on the penile smooth muscle of abnormal mucus type impotence syndrome rats, disease syndrome drug group rats and normal control group rats, to verify the protein in the tissue of the present invention Whether the candidate differential protein can be used as a drug target marker for Yimusak tablet in penile smooth muscle tissue of rats with abnormal mucus type impotence syndrome:
[0171] That is, protein No. 4 P02625 (parvalbumin α); protein No. 15 Q9Z2L0 (voltage-dependent anion-selective channel protein 1); protein No. 26 P47853 (bischain) and protein No. P04904 (glutathione S-transferase α-3); use the antibodies of the above proteins respectively (all antibodies are purchased from Abcam (Shanghai) Trading Co., Ltd.), and treat the normal control group, d...
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