"Investigating the similarities and differences between traditional Chinese medicine and western medicine", the advantages and disadvantages of traditional Chinese medicine and western medicine are compared. In fact, the multi-target of traditional Chinese medicine has the advantage of synergistic effect, which is the defect of western medicine, and the composition of western medicine has clear mechanism and clear advantages. It is a defect of Chinese medicine. Therefore, at present, research on Chinese medicine first clears the ingredients and clarifies the mechanism. The two articles "How to publish high score SCI for Chinese herbal compound" are the significance of research on the effective ingredients of traditional Chinese medicine from the perspective of scientific research. On December 1 this year, the Fudan University new drug motherwort project was transferred, 150 million yuan. In forwarding this news, Ji Bo said, "At least five years, the search for effective ingredients from Chinese medicine to develop new drugs is the mainstream. From monomer return to compound, the development of molecular prescriptions, estimated at least five years later." Everyone pays attention to the scope of the "mainstream" here, not only in the field of Chinese medicine research, but in the field of new drug development. The article shared today is published in the magazine of the boss magazine Cell, Artemisinins Target GABAα Receptor Signaling and Impair a Cell Identity. 2016. Cell . Just published article. What is artemisinin, a new drug? Old medicine? For chemical ingredients, it is an old medicine that has been used clinically (the benefits of finding new drugs from Chinese herbal medicines, clinical use, and toxic effects are controllable). For indications, it is a new drug.
We analyze this article from the perspective of scientific research.
Look at the background first, the background is generally reported.
1. Loss of beta cells in type I diabetes. The subdivision of this study is how to supplement beta cells.
2. Alpha cells can be transformed into beta cells.
3. Overexpression of Pax4 can convert α cells into β cells, and Pax4 can inhibit Arx in β cells. Inhibition of Arx alone can also convert alpha cells to beta cells.
4, Artemisia annua extract can treat type 1 diabetes. (There are two references.) The article suggests that the role of artemisinin in the treatment of diabetes or in islets has not been reported. I don't know if everyone thinks about it. This is the idea of ​​developing new drugs from Chinese herbal medicine. From the clinical use of Chinese herbal medicine, find effective monomers and develop new drugs.
Figure 1. A Cell-Line Model of Transcription-Factor-Mediated Transdifferentiation for Identifying Functional ARX Inhibitors
The cell model uses the mouse β cell line Min6, and even the cell is also studied with a cell line, and the islet primary is difficult to handle.
An ARX-inducible overexpression stable strain was constructed. Add DOX, ARX expression, insulin drop (alpha phenotype). Insulin is used as a test indicator (molecular index). (In order to use inducible type, it is estimated that it is afraid of over-expression for a long time, and the cells cannot return β from α. The latter screening drug needs to change α to β.)
This stable strain was used as a cell model, and insulin was used as a detection index. In the presence of DOX (alpha type), HCS screening was performed. Artemisinin was screened.
Here, HCS is a screening drug. If it is a screening function gene, it is enough to change the drug to a genetic manipulation tool. The cell model and test indicators are the same.
After screening for artemisinin, it was verified in the α cell line αTC1 cells. Artemisinin can induce insulin expression in αTC1 cells. In fact, the previous screening strategy can be changed to use insulin as a detection index in αTC1 cells for screening, which is more convenient. So for the embarrassing article to use the previous complex strategy, there must be a reason. There is no clear statement in the text.
In the end, it is the article of cell. In the first picture, the effect of drugs on the nuclear localization of ARX is also discussed, because ARX is a transcription factor.
Figure 2. Artemether Induces Insulin Expression in a Cells by Targeting Gephyrin
OK, the first picture got the medicine, because it is a functional screening, so the drug function is there ( pharmaceutical evaluation - cell experiment ). Let's start the mechanism below. It is noted that the screening drug is in Min6 cells, but the mechanism is discussed in αTC1 cells.
Fix the drug molecule and perform a pulldown experiment. Mass spectrometry identified the pulled down proteins. I found a protein that can bind directly to artemisinin, gephyrin. The drug target found here is a direct target. In fact, everyone looks at the description of the first picture, the author also did a dosing and non-medicine protein group, looking for differential proteins after the drug. Indirect targets can be found in these differential proteins.
Then, the drug target was identified. Using RNAi to knockdown gephyrin in αTC1 cells, it was found that the expression of insulin could not be upregulated after the addition of artemisinin. ( Drug and drug target correlation - functional correlation - recovery experiment ). In the end is the cell, directly on the test. F2D. In the F2E diagram, artemisinin was added and gephyrin rose ( drug-drug-related-molecular correlation ).
Figure 3. Artemether Increases GABA-Receptor Signaling in a Cells
I found the drug in the front and found the drug target. Here I started to explore the drug target mechanism. Everyone pays attention to the details. The title of the picture is written with artemisinin. The title of the article is gephyrin. Gephyrin's related experiments were placed in the supplemental data. The reason is very simple. Gephyrin's related experiments are not very innovative, and the signal pathway downstream of drugs and drug targets is a new relationship, which is innovative and puts the text. Because Gephyrin is associated with GABA receptors, this picture does a bunch of electrophysiology. Explain that drugs can alter downstream signaling molecules.
It is noted that drug intervention and drug target intervention, downstream detection, are not consistent for all downstream molecules. Because the drug may have other targets. This cell also did not show both data at the same time.
Figure 4. Hormone Secretion Controls Pancreatic Cell-type Stability
This article is mainly to explore the conversion of α and β. The fourth panel explores the effects of drugs on cell types. Artemisinin may inhibit the alpha phenotype.
Figure 5. Artemether Increases b Cell Mass In Vivo
The fourth picture shows that artemisinin may inhibit the alpha phenotype, so whether it can promote the beta phenotype. Directly on animal experiments. Looking at the experimental design, GFP was placed behind the glucagon promoter, and the number of α cells was observed by GFP. MCherry was placed behind the insulin promoter and mCherry was used to observe the number of beta cells. After the artemisinin treatment, the α cells were reduced and the β cells were increased (additional data). ( Evaluation of efficacy - animal experiment )
It is not enough to look at normal animals and build a disease model. Caspase-8 was constructed behind the insulin promoter. By adding a dimer, the beta cells are apoptotic. Simulate type 1 diabetes. Artemisinin was added to this model, and artemisinin was evaluated to increase beta cells based on the number of red cells.
Experiments were also performed on mice and it was found that artemisinin also increased beta cells and that the increased beta cells were derived from alpha cells. The experimental design is very clever.
Figure 6. Artemether Affects a Cell Identity in Human a Cells
The previous ones are all animal cells. The sixth picture is the use of human primary cells to explore the relationship between artemisinin and its downstream signaling molecules. Although a large number of protein groups and transcriptomes have been previously worked in animal cells, a large number of transcriptome work has been done in human cells.
Figure 7. Artemether Enhances Insulin Secretion in Human Islets
Finally, the effect of artemisinin on insulin secretion in human cells was discussed. ( Pharmaceutical evaluation - cell experiment )
Although this article is published on the cell, we can see that Gephyrin's related experiments are not much in terms of molecular mechanism. There is no detailed discussion of the relationship between Gephyrin and ARX. However, the highlight is the artemisinin drug, which converts between islet cells alpha and beta.
If this article, artemisinin and Gephyrin are not directly interacting, the quality of other data is not up to the cell requirements. However, it is difficult to find targets for direct interaction of drugs. In addition, generally this type of article is nature medicine, this issued a cell. Estimated to be the importance of the disease.
Chinese medicine emphasizes "yin and yang", and α and β in islets can be regarded as a pair of yin and yang. Insulin and glucagon are also a pair of yin and yang. To develop new drugs, you can look at the medicines from the prescriptions of Chinese medicine practitioners and find new drugs from the medicines. As for the five years later, how to develop new molecular formulas based on effective monomers, and to retain the advantages of multi-targets of traditional Chinese medicine compounds. Refer to the theory of Chinese medicine.
How to screen drugs and how to screen new functional genes. Refer to the idea of ​​the first picture in this article. Determine cell models and test indicators. The cell model must be a cell line of cell lines. The detection index can be the same as the use of molecular indicators, as well as behavioral indicators, such as proliferation and apoptosis. As for the platform, it is a high-throughput platform that Jibo often talks about. This platform was originally used to screen drugs, but now we have a ready-made gene manipulation library (overexpression, interference, cas9), which is used for new functional gene screening.
to sum up
First, the efficacy evaluation
Cell experiments: first in animal cell lines, then in human primary cells.
Animal experiments: zebrafish and mice.
Second, the drug mechanism - drug target identification
1. Clinical relevance: detection on animals.
2, drug target function (with the following function recovery experiment)
3. Correlation between drugs and drug targets
Molecular correlation: The drug up-regulates Gephyrin, mainly in direct interaction.
Functional relevance: Direct response to the experiment.
4. Drug target mechanism: GABA receptor and downstream signaling molecules.
PS: I hope that friends who have more exchanges with Jibo in learning and experimentation can pay attention to Jikai Gene WeChat and reply to the words “Jiboâ€.
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