| Neurodegenerative diseases such as Parkinson’s disease (PD), Alzheimer’s disease (AD), HIV-associated neurocognitive disorder and amyotrophic lateral sclerosis (ALS) are always being realized to have the chronic, progressive degeneration of the neuron in the central nervous system (CNS).Recent studies indicate that there is a particularly close relationship between the polarization of microglia and the neurodegenerative diseases. Studies of peripheral microglia have led to the concept of different activation states, ranging from’classical’ activation (M1 type) to so-called’alternative’activation (M2 type). The M1 type is relevant to responses to bacterial and viral infection; and the M2 type is associated with immune responses to parasites and tissue-repair programmes. In general, classically activated macrophages are most commonly associated with disease states that are driven by low-grade forms of inflammation, while alternative activation states are generally associated with protection from diseases in which classical activation is pathogenic.Therefore, these theories suggest that the anti-inflammatory drugs could inhibit the activation of microglia and protect against the disease. NSAIDs can significantly reduce the risk of the disease, while it has significantly side effects, such as gastrointestinal reaction, cardiovascular damaging, renal damaging and hypertension.Traditional Chinese medicines have abundant resources and good synergistic effect with small toxic side effects. Many of the active ingredients in traditional Chinese medicine is used to treat a variety of diseases. Among them, the anti-inflammatory medicine which generally refers to detoxify class, has a detoxification, has been used in some anti-inflammatory, anti-immune disorders.In recent years, though transcriptomics and metabolomics have developed rapidly, metabolomics has not yet effectively applied to characterize the different metabolites in cellular regulation, especially including microglia. Metabolomics have less type various and fewer quantity of metabolites, herein we use metabolomics method in this study to examine the effect of anti-inflammatory medicine monomers on microglia polarization.First, thirty-one candidate medicine monomer were selected and tested by Q-PCR, using M1 marker (interleukin-1beta, tumor necrosis factor-alpha and inducible nitric oxide synthase) and M2 markers (arginine synthase-1, FIZZ-1, CD206, IGF-1), to determine the effect on polarization of microglia. Among these candidates, we found Puerarin is an effective inhibitor of microglia Ml phenotype with low toxicity.Then metabolism differences on polarization in M1, M2 type and naive microglia were analyzed and compared using GC-MS and UPLC-Q-TOF-MS metabolomics method. In non-target studies,27 and 25 metabolites in 12 and 15 metabolic pathways are different between resting state and M1 or M2 type, respectively. Then, the fingerprint and footprint metabolism method is applied to study the regulation of Puerarin on microglial polarization. By metabolic profiling and multivariate statistical analysis, totally 42 kinds of different metabolites were screened out in the resting state, M1 and M2 type, including lipid metabolism, vitamin B1 metabolism, niacin and nicotinamide metabolism, and purine metabolism. We chose these metabolites as criteria to evaluate puerarin metabolites and discovered that puerarin could affect the switch of M1 to M2 phenotype in microglia.This study indicated Puerarin as a candidate to barricade microglial M1 activation, and define a new pattern to test drugs in neurodegenerative diseases. |
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