Tangshen formula targets the gut microbiota to treat non-alcoholic fatty liver disease in HFD mice: A 16S rRNA and non-targeted metabolomics analyses.

BACKGROUND: Tangshen formula (TSF) has an ameliorative effect on hepatic lipid metabolism in non-alcoholic fatty liver disease (NAFLD), but the role played by the gut microbiota in this process is unknown. METHOD: We conducted three batches of experiments to explore the role played by the gut microbiota: TSF administration, antibiotic treatment, and fecal microbial transplantation. NAFLD mice were induced with a high-fat diet to investigate the ameliorative effects of TSF on NAFLD features and intestinal barrier function. 16S rRNA sequencing and serum untargeted metabolomics were performed to further investigate the modulatory effects of TSF on the gut microbiota and metabolic dysregulation in the body. RESULTS: TSF ameliorated insulin resistance, hypercholesterolemia, lipid metabolism disorders, inflammation, and impairment of intestinal barrier function. 16S rRNA sequencing analysis revealed that TSF regulated the composition of the gut microbiota and increased the abundance of beneficial bacteria. Antibiotic treatment and fecal microbiota transplantation confirmed the importance of the gut microbiota in the treatment of NAFLD with TSF. Subsequently, untargeted metabolomics identified 172 differential metabolites due to the treatment of TSF. Functional predictions suggest that metabolisms of choline, glycerophospholipid, linoleic acid, alpha-linolenic acid, and arachidonic acid are the key metabolic pathways by which TSF ameliorates NAFLD and this may be influenced by the gut microbiota. CONCLUSION: TSF treats the NAFLD phenotype by remodeling the gut microbiota and improving metabolic profile, suggesting that TSF is a functional gut microbial and metabolic modulator for the treatment of NAFLD.

High-sulfated derivative of polysaccharide from Ulva pertusa improves Adriamycin-induced nephrotic syndrome by suppressing oxidative stress.

Nephrotic syndrome (NS) is characterized by proteinuria, hyperlipidemia, and hypoalbuminemia. Ulva pertusa, a green seaweed, is a nutritional supplement. In this study, the high-sulfated derivative of Ulva pertusa polysaccharide (HU) was prepared by combining U pertusa polysaccharide with chlorosulfonic acid. The NS rat model was established by tail vein single injection of Adriamycin (6.0 mg kg-1). Normal rats were used as the control group. NS rat models were treated with HU or U (173 mg kg-1 day-1). After treatment for 6 weeks, we assessed urine protein, renal function, and blood lipids, and observed morphology and histologic injury of the kidney and glomerular microstructure. Furthermore, we detected antioxidant enzyme activity and expression level of the Keap1/Nrf2 signaling pathway to explore the potential mechanism of HU. Results showed that HU not only alleviated hyperlipidemia and hypoalbuminemia, but also reduced urine protein by inhibiting podocyte detachment, thickening of the glomerular basement membrane, and expression of kidney fibrosis markers (collagens I and IV). In addition, HU enhanced antioxidant enzyme activity (GSH-Px, CAT, SOD) in both serum and the kidney, which may be due to upregulating the expression of Nrf2 and downregulating the expression of Keap1. In conclusion, HU appears to be effective in attenuating NS in rats through suppressing oxidative stress by regulating the Keap1/Nrf2 signaling pathway.

Characteristics of Serum Metabolites and Gut Microbiota in Diabetic Kidney Disease.

Disturbance of circulating metabolites and disorders of the gut microbiota are involved in the progression of diabetic kidney disease (DKD). However, there is limited research on the relationship between serum metabolites and gut microbiota, and their involvement in DKD. In this study, using an experimental DKD rat model induced by combining streptozotocin injection and unilateral nephrectomy, we employed untargeted metabolomics and 16S rRNA gene sequencing to explore the relationship between the metabolic profile and the structure and function of gut microbiota. Striking alterations took place in 140 serum metabolites, as well as in the composition and function of rat gut microbiota. These changes were mainly associated with carbohydrate, lipid, and amino acid metabolism. In these pathways, isomaltose, D-mannose, galactonic acid, citramalic acid, and prostaglandin B2 were significantly upregulated. 3-(2-Hydroxyethyl)indole, 3-methylindole, and indoleacrylic acid were downregulated and were the critical metabolites in the DKD model. Furthermore, the levels of these three indoles were restored after treatment with the traditional Chinese herbal medicine Tangshen Formula. At the genera level, g_Eubacterium_nodatum_group, g_Lactobacillus, and g_Faecalibaculum were most involved in metabolic disorders in the progression of DKD. Notably, the circulating lipid metabolites had a strong relationship with DKD-related parameters and were especially negatively related to the mesangial matrix area. Serum lipid indices (TG and TC) and UACR were directly associated with certain microbial genera. In conclusion, the present research verified the anomalous circulating metabolites and gut microbiota in DKD progression. We also identified the potential metabolic and microbial targets for the treatment of DKD.

Structural Characterization and Anti-Nonalcoholic Fatty Liver Effect of High-Sulfated Ulva pertusa Polysaccharide.

The high-sulfated derivative of Ulva pertusa polysaccharide (HU), with unclear structure, has better anti-hyperlipidmia activity than U pertusa polysaccharide ulvan (U). In this study, we explore the main structure of HU and its therapeutic effect against nonalcoholic fatty liver disease (NAFLD). The main structure of HU was elucidated using FT-IR and NMR (13C, 1H, COSY, HSQC, HMBC). The anti-NAFLD activity of HU was explored using the high-fat diet mouse model to detect indicators of blood lipid and liver function and observe the pathologic changes in epididymal fat and the liver. Results showed that HU had these main structural fragments: →4)-ß-D-Glcp(1→4)-α-L-Rhap2,3S(1→; →4)-α-L-Rhap3S(1→4)-ß-D-Xylp2,3S(1→; →4)-α-L-Rhap3S(1→4)-ß-D-Xylp(1→; →4)-α-L-IdopA3S(1→4)-α-L-Rhap3S(1→; →4)-ß-D-GlcpA(1→3)-α-L-Rhap(1→; →4)-α-L-IdopA3S(1→4)-ß-D-Glcp3Me(1→; →4)-ß-D-Xylp2,3S(1→4)-α-L-IdopA3S(1→; and →4)-ß-D-Xylp(1→4)-α-L-IdopA3S(1→. Treatment results indicated that HU markedly decreased levels of TC, LDL-C, TG, and AST. Furthermore, lipid droplets in the liver were reduced, and the abnormal enlargement of epididymal fat cells was suppressed. Thus, HU appears to have a protective effect on the development of NAFLD.

Tangshen Formula Improves Diabetes-Associated Myocardial Fibrosis by Inhibiting TGF-ß/Smads and Wnt/ß-Catenin Pathways.

Cardiovascular disease has become the main cause of death among complications of diabetes. Myocardial fibrosis is a crucial pathological change of cardiovascular disease. Tangshen Formula (TSF) shows a good clinical effect in the treatment of diabetic kidney disease (DKD). However, whether TSF alleviates diabetes-associated myocardial fibrosis is still unknown. In the present research, we studied the effect and mechanism of TSF in the treatment of myocardial fibrosis in vivo and in vitro. We found that TSF treatment significantly downregulates myocardial fibrosis-related markers, including collagens I and III, and α-SMA. TSF also protects primary mouse cardiac fibroblast (CF) from transforming growth factor-ß1- (TGF-ß1-) induced damage. Moreover, TSF decreased the expression levels of TGF-ß/Smad-related genes (α-SMA, collagens I and III, TGF-ß1, and pSmad2/3), and increased Smad7 gene expression. Finally, TSF decreased the expressions of wnt1, active-ß-catenin, FN, and MMP7 to regulate the Wnt/ß-catenin pathway. Taken together, TSF seems to attenuate myocardial fibrosis in KKAy mice by inhibiting TGF-ß/Smad2/3 and Wnt/ß-catenin signaling pathways.

Antioxidant and antihyperlipidemic activities of high sulfate content purified polysaccharide from Ulva pertusa.

In this study, purification of polysaccharide ulvan by anion exchange chromatography was prepared, and the major polysaccharide fraction (FU) was collected at 1.0 M NaCl elute by anion exchange chromatography, then high sulfate content purified ulvan (HFU) was prepared with sulfur trioxide/N,N-dimethylformamide (SO3-DMF) in formamide. The antioxidant activity and the antihyperlipidemic activity of HFU in mice were determined. The results showed that treatment with HFU could improve the antioxidant and antihyperlipidemic activity. Compared with the hyperlipidemic group, the antihyperlipidemic activity of HFU (125 mg/kg) was the strongest, TC concentrations were significantly decreased by 26.7% (P < .01), significantly reduced LDL-C (32.6%, P < .01), significantly increased HDL-C (19.6%, P < .01), and HFU-treated group (250 mg/kg) exhibited optimal effects on TG (29.0%, P < .01); the HFU groups at the doses of 125 mg/kg could significantly decrease the MDA (29.9%, P < .01); the HFU groups at the doses of 500 mg/kg could increase the activities of GSH-Px obviously (19.9%, P < .01).