The Antihypertensive Effects and Mechanisms of Huotan Jiedu Tongluo Decoction in Rats with H-Type Hypertension
May 17th, 2024
Here, we present a protocol to induce H-type hypertension and evaluate the antihypertensive effects of Huotan Jiedu Tongluo decoction (HTJDTLD) administered intragastrically. In rats with H-type hypertension, HTJDTLD had effective antihypertensive effects, possibly associated with inhibition of endoplasmic reticulum (ER) stress-induced apoptosis pathway activation.
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.
Overview
This study presents a protocol to evaluate the antihypertensive effects of Huotan Jiedu Tongluo decoction (HTJDTLD) in rats with H-type hypertension. The findings suggest that HTJDTLD effectively reduces hypertension, potentially by inhibiting ER stress-induced apoptosis.
Key Study Components
Area of Science
- Neuroscience
- Pharmacology
- Traditional Medicine
Background
- H-Type hypertension is a significant public health issue.
- Current treatments primarily rely on western medicine.
- Traditional Chinese medicine offers alternative therapeutic options.
- Understanding the mechanisms of these treatments is crucial for effective management.
Purpose of Study
- To explore the antihypertensive effects of HTJDTLD.
- To investigate the underlying mechanisms of action.
- To identify potential biomarkers for diagnosis and prognosis.
Methods Used
- Induction of H-type hypertension in rat models.
- Administration of HTJDTLD via intragastric route.
- Evaluation of blood pressure changes.
- Assessment of ER stress and apoptosis pathways.
Main Results
- HTJDTLD demonstrated significant antihypertensive effects in rats.
- The mechanism may involve inhibition of ER stress-induced apoptosis.
- Potential for HTJDTLD as a treatment option for H-type hypertension.
- Future research will focus on biomarkers and therapeutic strategies.
Conclusions
- HTJDTLD is effective in managing H-type hypertension in rats.
- Inhibition of ER stress pathways is a key mechanism.
- Further studies are needed to explore clinical applications.
Frequently Asked Questions
Executive Industry Relevance
H-type hypertension, marked by elevated homocysteine, presents a mechanistically complex challenge for early-stage cardiovascular drug discovery. The protocol enables quantitative assessment of antihypertensive interventions and mechanistic de-risking via ER stress pathway interrogation. This supports predictive confidence in target validation and informs portfolio triage for vascular disease programs.
Strategic Applications in Biopharma R&D
Early Discovery & Target Validation
- Enables interrogation of ER stress-induced apoptosis as a mechanistic driver in hypertension models.
- Supports functional target validation by quantifying gene and protein expression changes in GRP78, JNK, TRAF2, and caspase-3.
- Facilitates predictive confidence in pathway modulation for cardiovascular indications.
Screening & Assay Development
- Provides a validated in vivo system for evaluating antihypertensive compound efficacy.
- Standardizes quantitative blood pressure and homocysteine measurements for reproducible screening outputs.
- Enables downstream assay development for ER stress and apoptosis pathway readouts.
Translational & Preclinical Research
- Aligns preclinical biomarker measurement (homocysteine, ER stress markers) with disease-relevant endpoints.
- Supports continuity from discovery through preclinical validation in vascular disease models.
- De-risks advancement decisions by linking mechanistic inhibition to functional outcomes.
Pipeline & Workflow Integration
This protocol positions mechanistic hypertension models at the interface of early discovery and preclinical validation, enabling hypothesis-driven target evaluation and quantitative efficacy assessment.
- Discovery Biology: Supports hypothesis testing for ER stress pathway involvement in hypertension.
- Screening: Delivers reproducible, quantitative blood pressure and biomarker outputs for compound evaluation.
- Analytics: Integrates ELISA, qRT-PCR, and western blot data for robust statistical comparison of intervention effects.
- Translational Research: Aligns preclinical biomarker changes with disease-relevant mechanistic endpoints.
- Enterprise Reuse: Provides a reusable in vivo platform for cardiovascular and vascular target programs.
Operational & Enterprise Impact
- Scientific Value: Increases predictive confidence in target and pathway selection for hypertension therapies.
- Operational Value: Standardizes in vivo and molecular readouts for cross-study reproducibility.
- Strategic Value: Informs go/no-go decisions by linking mechanistic inhibition to functional blood pressure outcomes.
- Portfolio Impact: Enables risk-adjusted prioritization of cardiovascular candidates based on mechanistic and phenotypic data.
Implementation Considerations
- Requires expertise in in vivo hypertension models and molecular assay execution.
- Demands access to noninvasive blood pressure measurement, ELISA, qRT-PCR, and western blot instrumentation.
- Necessitates cross-team standardization of biomarker and blood pressure protocols.
- Adaptation to other disease models may require pathway-specific validation.
- Interpretation limited to mechanistic and preclinical endpoints supported by the protocol.
Biopharma FAQs
Chapters in this video
0:00
Introduction
1:08
Treatment of Methionine‐Induced H‐Type Hypertensive Rats with Huotan Jiedu Tongluo Decoction (HTJDTLD)
3:33
Plasma Homocysteine (Hcy) ELISA Measurement of Blood from H‐Type Hypertensive Rats Treated with Huotan Jiedu Tongluo Decoction
