Description

Background

Type 2 diabetes accounts for 90% of all kinds of diabetes caused by a declining response to insulin, the victims suffer from huge side effects. And the number of cases is increasing rapidly. Moreover, it is gradually spreading among younger crowds. Therefore, we find it helpful to many people to design a drug used to prevent symptoms of type 2 diabetes.

Throughout this experiment, our team has applied skills regarding lab technique and team synergy. In order to formulate an experiment to match the general preferences of the people, we first divide the topics into three categories: food, industry, and medicine. We distributed questionnaires to many people, and the results showed that everyone was more curious about medicine. We realize that we need to study the problem in this direction.

Experiment design

FoxO1

According to published research, we found that Forkhead box protein O1 (FoxO1) plays an important role in initiating the transcription of enzymes G6Pace and PEPCK. Thus, we intend to reduce the expression of the two enzymes by regulating the expression of FoxO1. After confirming our topic, we listed the experimental procedures, which were refined further in discussion and failures, in detail.

General experiment procedure

We transfected HepG2 cells with plasmids containing hFoxO1, and the IRE sequence could be combined with hFoxO1 and initiate the transcription of downstream genes. In order to test the expression level of downstream G6Pace and PEPCK, we replaced them with the luciferase gene. Then we added the substrate of the enzyme to test its activity of the enzyme.

Expect results

To regulate the expression of hFoxO1, we will use small molecular compounds from the database to examine their effects on gluconeogenesis. Small molecule compounds with inhibitory activity of FoxO1 transcriptional activation will be screened by reporter gene assay. We will further study the compound's inhibitory effect on gluconeogenesis in hepatocytes and the effect on the mRNA level of key enzymes of gluconeogenesis in hepatocytes to determine the hypoglycemic effect of the compound. The small-molecule compounds screened in this work will be used as drugs for the treatment of type 2 diabetes in the future.

Reference

IDF., Diabetes Atlas. 10th Edition, 2021.
Choi, H.E., et al., Novel FoxO1 inhibitor, JY-2, ameliorates palmitic acid-induced lipotoxicity and gluconeogenesis in a murine model. Eur J Pharmacol, 2021. 899: p. 174011.
Oh, K.J., et al., CREB and FoxO1: two transcription factors for the regulation of hepatic gluconeogenesis. BMB Rep, 2013. 46(12): p. 567-74.
Zhao, Y., Y. Wang, and W.G. Zhu, Applications of post-translational modifications of FoxO family proteins in biological functions. J Mol Cell Biol, 2011. 3(5): p. 276-82.
Nagashima, T., et al., Discovery of novel forkhead box O1 inhibitors for treating type 2 diabetes: improvement of fasting glycemia in diabetic db/db mice. Mol Pharmacol, 2010. 78(5): p. 961-70.
Ohtake, F., et al., Modulation of oestrogen receptor signalling by association with the activated dioxin receptor. Nature, 2003. 423(6939): p. 545-50.