Application of doxepin in the preparation of drugs for preventing or treating metabolic diseases caused by insulin
A doxepin, hyperinsulinemia technology, applied in metabolic diseases, drug combinations, endocrine system diseases, etc., can solve problems such as body fluid retention and weight gain
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Embodiment 1
[0051] Example 1 Drug Action Mechanism Research
[0052] Experimental materials: human liver cancer cell line HepG2 (purchased from ATCC, USA)
[0053] Culture conditions: Human liver cancer cell line HepG2 was cultured in DMEM (HG) medium at 37°C, CO 2 in an incubator with 5% content and 50% humidity.
[0054] 1.1 Detection of the dose effect of doxepin on the activation of FMA3A
[0055] The above-mentioned cells were subcultured in six-well plates, starved for 12 hours, and treated with doxepin (Dr. μM rosiglitazone (Sigma, R2408) was used as a positive control, and 0 μM doxepin was used as a negative control. Cells were collected after 48 hours of treatment.
[0056] The cells collected above were divided into two parts, one part was lysed with 1x lysis buffer (Reporterlysis buffer), and proteins were extracted for western blot respectively; the other part was extracted with Trizol kit (BioTeke, RP1202), and the RNA was reverse transcribed to make RT-PCR.
[0057] Exp...
Embodiment 2
[0072] The preparation of embodiment 2 obesity animal model
[0073] Purchase 8 8-week-old C57BL mice, and start high-fat feeding after adapting to a feeding environment with 50-60% humidity at 24°C for 3 days. Feed for 12 weeks. Monitor fasting blood glucose, plasma triglycerides and cholesterol, triglycerides and cholesterol in the liver, and body weight.
[0074] 2.1 Determination of fasting blood glucose in mice
[0075] The mice in the high-fat group and the control group were fasted for 6 hours and drank water freely, which was recorded as the 0min point (the 0 point is the fasting blood glucose point before glucose was given after 6 hours of starvation), and then 5 mg / g of body weight glucose was given for intragastric administration, and at the same time Start timing, take blood from the tail vein in a quiet state at 30min, 60min, 90min, and 120min respectively, and measure fasting blood glucose with a blood glucose meter. Wherein, the body weight of the mice was me...
Embodiment 3
[0086] Embodiment 3 pharmacozoological experiment
[0087] After the successful establishment of the mouse obesity animal model, the mice were divided into the following four groups:
[0088] The control diet group was fed with feed containing 10% fat and treated with normal saline;
[0089] The high-fat diet negative control group was fed with 45% fat feed and treated with normal saline;
[0090] The high-fat diet doxepin group was fed with feed containing 45% fat, treated with doxepin at a dose of 10 mg / kg.day, and intraperitoneally injected;
[0091] The high-fat diet rosiglitazone group was fed with feed containing 45% fat, treated with rosiglitazone at a dose of 15 mg / kg.day, and intragastrically administered.
[0092] 3.1 Determination of mouse body weight
[0093] The above four groups of mice were treated continuously for one month, and the body weight changes of each group of mice were measured once a week, and the measurement results were as follows: Figure 7 A ...
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