H-Type hypertension has become a significant public health challenge of human. It mainly relies on western medicine treatment, which can have said effects. Our research explores the anti-hypertensive effects and mechanisms of Huotan Jiedu Tongluo Decoction, a traditional Chinese medicine formula in rats with H-Type hypertension.
Our study indicated said Huotan Jiedu Tongluo Decoction had anti-hypertensive effect in rats with H-Type hypertension. The underlying mechanism may involve inhibiting ER stress activation-induced apoptosis which provide an effective treatment option for patients. We will explore the potential biomarkers for early diagnosis and prognosis and develop novel therapeutic strategies for H-type hypertension in the future.
To begin, segregate 50 hypertensive rats into five groups including control. Give the rats in group one methionine, or met group, a diet of 3%methionine for 28 days to induce H-Type hypertension. For group 2 to 4 rats, administer a combination of Huotan Jiedu Tongluo Decoction and enalapril maleate in addition to the methionine diet for 28 days.
Next, create a restraining device by placing a cylindrical restraint mesh into a thermal tube and putting the thermal tube into a canvas cover. After 28 days of treatment, place a rat in the mesh. Next, place the signal cable of a non-invasive sphygmomanometer under the cover.
Position the rat's tail in the gap in the cover and secure it on the stabilizing form pad. Transfer the rat to a quiet measurement site, 20 to 30 minutes before blood pressure measurement. Now connect the pressurized sensors'air hose connection, signal connection, and holding tube.
Place the pressurization sensor at the tip of the rat's tail. A pulse wave should appear after the rat tail is inserted into the sensor. Press start/stop to initiate or stop the measurement.
When the blood pressure test is completed, the result menu will pop up automatically. Check the average value of the measurement, standard deviation, standard, and coefficient of variation, in the results menu. After euthanizing the rat, dissect it with a pair of surgical scissors and tooth forceps from the perineum to the neck.
Turn over the thoracic and abdominal contents. Then navigate to the abdominal aortic bifurcation between the two kidneys. Collect the aorta up to a section of the aortic arch.
The systolic and diastolic blood pressures were significantly greater in the met group relative to the control group from one to four weeks. The blood pressures decreased after the administration of the Huotan Jiedu Tongluo Decoction. The combined effect of the decoction and/or enalapril maleate had a stronger anti-hypertensive effect than the decoction alone.
To begin, centrifuge the coagulated blood-containing tubes at 626 to 1, 409 G for 20 minutes at two to eight degrees Celsius. Then carefully collect the supernatant in a new tube. Store the supernatant at 80 degrees Celsius.
Next, set up blank, standard, and sample wells. Pipette 50 microliters of the diluted standards into the ELISA plate then transfer 40 microliters of the diluted samples into the sample wells. Add 10 microliters of serum into the sample wells.
Now, seal the plate with a sealing film. Place the sealed plate in an incubator at 37 degrees Celsius for 30 minutes. Dilute the 30x concentrated wash solution with distilled water to obtain a single-strength wash solution.
Next, remove the ceiling film from the plate and discard the liquid. Now, fill each well with 200 microliters of washing solution five times. After discarding the supernatant for the last time, tap the plate dry.
Pipette 50 microliters of the enzyme reagent into all wells other than the blank ones. Then seal the plate with a ceiling film before placing it in an incubator at 37 degrees Celsius for 30 minutes. After washing the plate five more times, add 50 microliters of each of the color developers A and B.Shake the plate gently to mix well, then incubate the plate at 37 degrees Celsius at low light for 10 minutes.
Now, pipette 50 microliters of the stop solution into each well to terminate the reaction. Measure the absorbance of each well sequentially at 450 nanometers. The plasma homocysteine concentration in the methionine group was twofold greater than in the control group.
The plasma homocysteine concentrations were relatively lower in the treatment groups.
