Gene-Level Stress Assessment of Cardiomyocyte
Some drugs and environmental pollutants are considered to have potential myocardial cytotoxicity, which may seriously lead to the risk of cardiovascular diseases such as heart failure, arrhythmia, cardiac hypertrophy, and congestive heart failure. To explore the mechanism, in addition to the assessment of myocardial cytotoxicity, sometimes it is necessary to study the signal pathways at the gene level.
To explore the relationship between target genes and oxidative stress, Creative Bioarray has developed a one-stop experimental platform. First, according to the requirements of customers, simulate the environment of cellular oxidative stress, and then study the expression changes of target genes based on the cell model. This service is expected to help customers explore the interaction between cell oxidative stress and specific genes.
Service Case
S is a ubiquitous substance in the environment. Studies have shown that long-term exposure to S can cause cardiovascular and cerebrovascular diseases, abnormal embryonic development, and so on. However, S is also an effective drug in the clinic. More and more studies have proved that oxidative stress is one of the important mechanisms of S-induced cytotoxicity. Gene Y is a key protein that regulates oxidative stress and plays an important role in mitochondrial ROS metabolism and apoptosis. However, whether gene Y plays an important role in the mechanism of S-induced oxidative stress in cardiomyocytes remains to be explored.
Experimental scheme
To help customers explore whether gene Y is involved in S-induced oxidative stress in cardiomyocytes, we knocked down and overexpressed gene Y in cardiomyocytes H9c2, and established a cell oxidative stress model with S. By observing the changes in intracellular ROS levels in cardiomyocytes treated with s, to explore whether gene Y can receive S to induce the increase of ROS level in cardiomyocytes.
Service Content
We transfected the myocardial cell lines provided by customers with plasmids containing siRNA and Y gene overexpression respectively to obtain knockdown and overexpression cell lines. Logarithmic long-term cells were digested with trypsin and then suspended. After inoculation and culture, different concentrations of drug S were added.
- The cell survival rate was detected by the CCK-8 method, the absorbance was detected by a microplate reader and the cell survival rate was calculated.
- Relationship between intracellular ROS level and substance S concentration detected by dichlorofluorescein (DCF) method.
- The relationship between the concentration of substance S detected by WB and the expression and phosphorylation of the Y gene.
- Localization of Y gene in cells confirmed by cellular immunofluorescence.
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