Metabolomics helps study Pu-erh tea protects mice from CRD by regulating tryptophan metabolism

Baiqu Metabolomics Information: @stay up all night young people's metabolic disorder first aid kit - drink Pu'er tea!
Title of the article: Pu-erh Tea Restored Circadian Rhythm Disruption by Regulating2 Tryptophan Metabolism
Published Journal: Journal of Agricultural and Food Chemistry
Impact Factor: 5.
895
Cooperative Unit: Southwest University Baiqu
Bio provides services: Tryptophan Metabolism

Research Background
Tea as a kind of Functional drinks, rich in polyphenols and amino acids, have the potential to regulate and improve circadian rhythm disorder CRD
.
Therefore, Pu-erh tea, as a daily tea drink, can be used as one of the optional substitutes for CRD prevention and treatment
.
The study found that Pu-erh tea extract could alleviate the glucose lipid and nitrogen metabolism in CRD-induced mice by regulating bile acid-mediated enterohepatic circulation
.
However, it is not enough to assess whether Pu-erh tea can relieve dizziness, lethargy and mental depression after staying up late
.
In recent years, tryptophan metabolism has shown potential to improve neurodegenerative and metabolic diseases
.
Therefore, the aim of this study was to elucidate whether Pu-erh tea protects healthy mice from CRD by regulating tryptophan metabolism
.


Research methods
Mice were randomly divided into W group (drinking water group) and T group (drinking 0.
25% Pu'er tea)
.
The W group and T group were randomly divided into four groups: CK-T, CK-W, CRD-T and CRD-W, and the illumination time of different groups was shown in Figure 1
.
The mouse serum, tissue, and fecal samples were collected for detection
.

 
Experimental results
01 Pu-erh tea alleviates CRD-induced damage to metabolism Pu-
erh tea slowed the increase in serum total bile acid content caused by CRD and increased the metabolism of bile acid in feces (Fig.
2A,B)
.
This suggests that CRD-treated mice have abnormal nitrogen metabolism, and Pu-erh tea can reduce lipid deposition by promoting bile acid metabolism
.
In addition, Pu-erh tea also restored CRD-induced liver rhythm gene oscillations (Fig.
2C–F)
.

02 Pu-erh tea improves brain signaling in
CRD-treated mice There was no rhythmic expression of neurotransmitters (GABA, dopamine, and 5-HT) in the brains of CRD-treated mice (Fig.
3A-C)
.
Analysis of rhythm gene expression results by qPCR indicated that Pu-erh tea regulated the oscillation of Bmal1 and Per1 genes, and up-regulated the trend of CRD-attenuated Clock and Cry1 genes (Fig.
4D–G)
.
Furthermore, CRD significantly increased IDO enzymatic activity in the brain (Fig.
5I) and accelerated tryptophan metabolism
.
Whereas high tryptophan depletion in the brain (Fig.
5H) accelerated the accumulation of KYNA and QUIN in the brain (Fig.
5J,K), increasing the risk of brain inflammation
.
Compared with the drinking water group (CK-W, CRD-W), the content of tryptophan in the Pu-erh tea group (CK-T, CRD-T) was significantly increased, which significantly inhibited the abnormal tryptophan metabolism induced by CRD
.

Figure 3.
The release of GABA, dopamine and serotonin in the brain of mice in different treatment groups at different time periods
 
03 Pu-erh tea reduced intestinal oxidative stress-mediated inflammation in CRD-treated mice.

CRD resulted in the formation of reactive oxygen species (ROS) in the gut and increased expression of its related proteins NOX2 and NOX4 (Fig.
6A,B)
.
Pu-erh tea reduced reactive oxygen species by up-regulating the expression of antioxidant pathway protein (Nrf2/HO-1)
.
With the reduction of ROS, the intestinal inflammatory pathway was significantly down-regulated, thereby protecting the intestinal tissue (Fig.
6C,D)
.

04 Pu-erh tea remodeled the gut microbiota of CRD-treated mice with
no significant changes in species richness between treatments (Fig.
7A)
.
ANOSIM-based principal component analysis (PCA) revealed distinct clusters of gut microbiota in each group (Fig.
7B)
.
There was a partial crossover between the CRD-T and CRD-W groups, which was significantly separated from the CK group, indicating that CRD significantly interfered with the composition of gut microbes
.
This conclusion was also confirmed by the results of NMDS analysis based on Bray-Curtis distance (Fig.
7C)
.

05 Pu-erh tea altered microbial metabolites in CRD-treated mice
SCFA of mice in the CK group showed circadian changes (Fig.
8A–F)
.
Peaks for isobutyric acid, isovaleric acid, and valeric acid occurred at 4 hours after the lights were turned on, while the lowest values ​​for acetic acid, propionic acid, and butyric acid occurred at 16 hours
.
Except for propionic acid, the content of CK-T group was higher than that of CK-W group, indicating that Pu-erh tea increased the production of SCFAs in vivo
.
CRD disrupted SCFA production
.
In particular, the contents of acetic acid, isobutyric acid, isovaleric acid and valeric acid were significantly reduced, and their rhythms were lost after CRD treatment
.
Pu-erh tea promoted short-chain fatty acid production and improved rhythmic oscillations in CRD-treated mice
.
Isobutyric acid, butyric acid, isovaleric acid, and valeric acid were most affected by Pu-erh tea, and their peak values ​​were comparable to or even higher than CK-T, but a rhythmic phase shift was observed
.


The tryptophan metabolism results showed that all samples had obvious aggregation (Fig.
9A,B)
.
The OPLS-DA model was validated using 200 displacement tests and no overfitting occurred (Fig.
9C,D)
.
We then screened the top 10 metabolites with VIP>1 for the main differential metabolites analyzed (Fig.
9E,F)
.
The CRD-W group significantly increased the concentrations of 3-HK, KYN, KYNA and Xa (Fig.
8G)
.
Compared with drinking Pu-erh tea (CRD-T group), the content of IAA, indole and 5-HT in Pu-erh tea increased even more than that of CK-W group
.
Compared with the CK-W group, the CK-T group increased the concentrations of IE, tryptamine and tryptophan in fecal metabolites
.
In other words, CRD promotes the kynurenine pathway in tryptophan metabolism
.
The intake of Pu-erh tea increased tryptophan and promoted the metabolism of tryptophan to indole pathway and serotonin pathway
.

Summary
CRD treatment poses threats to healthy mice, including disruption of glucose and lipid metabolism, intestinal oxidative stress and inflammation, and circadian clock instability
.
Pu-erh tea increases the indole and 5-HT pathways of tryptophan metabolism
.
Its metabolites (IAA, IE, indole, and 5-HT) further affect hepatic glucolipid metabolism and intestinal inflammatory pathway activation (MyD88/NF-κB) by interfering with hormone release (LEP, GC, GABA, and DA) in the brain )
.
Tryptophan metabolites and bile acids also promote hepatic lipid metabolism and inhibit intestinal oxidative stress through enterohepatic circulation
.
The findings underscore the importance of gut-liver-brain axis interaction in CRD regulation, where increased 5-HT metabolism of indole and tryptophan may be a therapeutic target for Pu-erh tea
.
In short, these data suggest that 0.
25% (w/v) Pu-erh tea (a human equivalent dose of 7 g/60 kg/day) has the potential to prevent CRD by modulating indole and 5-HT pathways
.


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