The function of nerve cells and brain tissue is highly dependent on mitochondrial function. As the main pharmacodynamic component of a variety of traditional Chinese medicines and compounds, lamp ethyl has been proven to have good pharmacological activity and clinical effect in cardiovascular and cerebrovascular diseases. **The mechanism and direct target of the neuroprotective effect of bulb B are undoubtedly of great value for guiding the rational use of drugs in clinical practice.
The Institute of Materia Medica, Chinese Academy of Medical Sciences is inadvanced science（if15.1) published an article titled "Scutellarin Rescued Mitochondrial Damage through Ameliorating Mitochondrial Glucose Oxidation via the PDK-PDC Axis". This study is the first to find that bulb B can regulate mitochondrial aerobic metabolism by targeting the mitochondrial pyruvate dehydrogenase kinase-pyruvate dehydrogenase complex axis (PDK2-PDC) in brain tissue, thereby exerting neuronal cell protection.
Findings:
1.Based on the analysis of energy metabolism in live cells, it was found that bulbylin had a regulatory effect on mitochondrial aerobic glucose metabolism
Fig.1 Lamp B can alleviate the pathological changes caused by chronic cerebral ischemia, improve cognitive dysfunction, and reduce oxidative stress.
Fig.2 Bulb B alleviates CCH-induced mitochondrial dysfunction.
2.Subcellular level mitochondrial proteomics and 13C isotope-labeled metabolic flux tracing studies were conducted to clarify the metabolic flow direction and flux regulated by bulb B, as well as the metabolic enzymes regulated
Fig.3 Mitochondrial proteomics studies revealed the regulatory effect of bulb B on oxidative phosphorylation and energy metabolism pathways.
Fig.4 The results of 13C MFA and Western blotting showed that bulb B had a regulatory effect on pyruvate metabolism on the glucose-pyruvate-TCA cycle axis.
3.Bulb B directly targets the PDK2-PDC axis to regulate mitochondrial aerobic metabolism
Fig.5 Kinase activity analysis showed that bulb B had obvious inhibitory activity against PDK2.
Fig.6 Molecular docking showed that brevityl could bind to the three pockets of PDK2.
Fig.7 Co-IP, DARTS and CESA experiments confirmed the interaction between bulbyl and PDK2.
Fig.8 Lip-MS experiments showed that there was a direct binding pocket between bulb B and PDK2.
Summary and discussion
Importance: This study provides an example of a small molecule targeting metabolic pathway for neurological diseases.
ChallengingThe identification of small molecule targets includes labeled and non-labeled methods, and it is difficult to choose the appropriate technology to improve the efficiency of target protein identification.
ExtensibilityIn view of the multi-target and multi-action pathways of traditional Chinese medicine, it is of great value to identify more target proteins of bulb B for the study of the mechanism and mechanism of related diseases.
Recommended by Zhongke Youpin
The focus and difficulty of monomer research is to find the target of its direct action. The probe method is one of the main methods for the identification of direct-acting targets. In this method, the drug is made into a probe and immobilized on a carrier, and then the protein bound to it is fished for identification. However, the probe method is limited by the experimental conditions of chemical synthesis and the structural properties of the drug, and its scope of application is limited. Some new chemical proteomic methods such as:Limited enzyme digestion (LIP-MS)., by analyzing the enzyme digestion status of the protein, whether it is changed due to drug binding, and large-scale identification of the bound protein can be realized. This type of methodThere is no need to prepare small molecules into probes, and it also provides information about the potential binding sequence of small molecules to targets。A schematic diagram of the basic principles of the chemical proteome by finite digestion is as follows:
On this basis, the affinity and binding kinetic constant were determined by combining surface plasmon resonance (SPR) and other techniques, and the functional mediating effect of the target was confirmed by functional testing, and the functional target directly acted on by the monomer could be finally confirmed. The research route is as follows:
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