Using UPLC-QTOF/MS and multivariate analysis to explore the mechanism of Bletilla Striata improving PM2.5-induced lung impairment.

Exposure to fine particulate matter (PM2.5) is closely related to lung diseases and has become more and more harmful to public health. The traditional Chinese medicine of Bletilla Striata has the effect of clearing and nourishing the lungs in clinics. The purpose of the study is using metabolomics methods to explore the mechanism of PM2.5-induced lung injury and Bletilla Striata's therapeutic effect. In this article, we used an Ultra Performance Liquid Chromatography-Quadrupole Time-of-Flight Mass Spectrometry (UPLC-QTOF/MS) method to identify the potential biomarkers. The results showed that there were 18 differential metabolites in the plasma and urine of rats with PM2.5-induced lung injury, involving the glycerophospholipid metabolism pathway, the tryptophan metabolism pathway, and the purine metabolism pathway, etc. After the administration, Bletilla Striata changed the levels of 21 metabolites, and partly corrected the changes in the level of metabolites caused by PM2.5. The results indicated that Bletilla Striata could exert a good therapeutic effect by reversing the levels of some biomarkers in the rats with PM2.5-induced lung impairment.

Efficient transfected liposomes co-loaded with pNrf2 and pirfenidone improves safe delivery for enhanced pulmonary fibrosis reversion.

Pulmonary fibrosis (PF) is an interstitial lung disease with complex pathological mechanism, and there is currently a lack of therapeutics that can heal it completely. Using gene therapy with drugs provides promising therapeutic strategies for synergistically reversing PF. However, improving the intracellular accumulation and transfection efficiency of therapeutic nucleic acids is still a critical issue that urgently needs to be addressed. Herein, we developed lipid nanoparticles (PEDPs) with high transfection efficiency coloaded with pDNA of nuclear factor erythroid 2-related factor 2 (pNrf2) and pirfenidone (PFD) for PF therapy. PEDPs can penetrate biological barriers, accumulate at the target, and exert therapeutic effects, eventually alleviating the oxidative stress imbalance in type II alveolar epithelial cells (AECs II) and inhibiting myofibroblast overactivation through the synergistic effects of Nrf2 combined with PFD, thus reversing PF. In addition, we systematically engineered various liposomes (LNPs), demonstrated that reducing the polyethylene glycol (PEG) proportion could significantly improve the uptake and transfection efficiency of the LNPs, and proposed a possible mechanism for this influence. This study clearly reveals that controlling the composition ratio of PEG in PEDPs can efficiently deliver therapeutics into AECs II, improve pNrf2 transfection, and synergize with PFD in a prospective strategy to reverse PF.

AAV induces hepatic necroptosis and carcinoma in diabetic and obese mice dependent on Pebp1 pathway.

Obesity and diabetes are risk factors for hepatocellular carcinoma (HCC); however, the underlying mechanisms are yet to be elucidated. Adeno-associated virus (AAV) frequently infects humans and has been widely used in gene therapy, but the risk of AAV infection such as HCC should be further evaluated. Here, we show that recombinant AAV injection caused liver injury, hepatic necroptosis, and HCC in db/db or high-fat diet-induced hyperglycemic and obese mice, but not in mice with only hyperglycemia or obesity. Prednisone administration or knockdown of Pebp1, highly expressed in db/db mice, alleviated hepatic injury and necroptosis induced by recombinant AAV in mice with diabetes and obesity. Inhibition of Pebp1 pathway also attenuated inflammation and necroptosis in vitro. Our findings show that AAV infection is a critical risk factor for HCC in patients with diabetes and obesity, and AAV gene therapy for these patients should be carefully evaluated. Both prednisone treatment and targeting Pebp1 pathway are promising strategies to alleviate inflammation and necroptosis that occurred in AAV gene therapy or related diseases.

Sustained Inflammation Induced by LPS Leads to Tolerable Anorexia and Fat Loss via Tlr4 in Mice.

Background: Sustained inflammation is implicated in a variety of pathological conditions like infection, obesity and type 2 diabetes. Lipid metabolism is crucial to support immune response during infection of bacteria. However, how sustained inflammation affects lipid metabolism, especially in white adipose tissue remains largely unknown. Methods: Sustained inflammation was induced by daily injection of Lipopolysaccharide (LPS). Tlr4 knockout mice were used to study the mechanism. Inflammation and lipid metabolism were evaluated by quantitative PCR, white blood cell counting, nuclear magnetic resonance, fat cell size quantification, lipolysis and fatty acid uptake assays, respiratory exchange ratio, and energy expenditure. Results: Here, we found that sustained inflammation leads to fat loss in mice with a quick loss and gradual increase manner. Moreover, LPS injection leads to inflammation, anorexia, decreased lipid anabolism, and increased lipid catabolism. Mechanically, we show that LPS induces fat loss, inflammation, anorexia, and alteration of lipid metabolism mainly dependent on Tlr4. Interestingly, sustained inflammation induces less fat loss, especially in epididymal white adipose tissue, than pair-feeding, and pair-feeding has no significant effect on inflammation and leads to less fatty acid uptake, more lipid catabolism and energy expenditure than LPS injection. In addition, we demonstrate that short-term sustained inflammation leads to relative long-term tolerance for LPS-induced anorexia, inflammation and altered lipid metabolism. Conclusion: These findings demonstrate that sustained inflammation induced by LPS leads to tolerable anorexia and fat loss via Tlr4 in mice, and provide new insights into the effect of sustained inflammation on lipid metabolism and subsequent tolerance.

Metabolomics analysis of Semen Cuscutae protection of kidney deficient model rats using ultra high-performance liquid chromatography-quadrupole time-of-flight Mass Spectrometry.

The traditional Chinese medicine syndrome "Kidney yang deficiency" is a kind of chronic kidney disease. With the development of society, the incidence of chronic kidney disease is increasing year by year, which also brings great economic pressure to people. Semen Cuscutae is an important traditional Chinese medicine to tonify liver and kidney, mainly used to tonify deficiency of liver and kidney, spleen and kidney deficiency and diarrhea. Although there are a lot of research at the molecular and cellular level to study the Semen Cuscutae on the treatment of Kidney yang deficiency syndrome, but there's no comprehensive research complete with metabolomics method from plasma, feces and urine metabolites aspects. The purpose of this study is to find the potential differential biomarkers of the Kidney yang deficiency model and blank group rats in plasma, urine and feces, and to investigate the mechanism of Semen Cuscutae in the treatment of Kidney yang deficiency syndrome. In this study, ultra high-performance liquid chromatography-quadrupole time-of-flight Mass Spectrometry (UPLC-QTOF/MS) was used to identify potential biomarkers. Through the analysis of metabolic profiles of plasma, urine, and feces, as well as multivariate statistical analysis and pathway analysis, the therapeutic mechanism of Semen Cuscutae for Kidney yang deficiency syndrome was described. The results showed that there were 69 differential metabolites in plasma, 93 differential metabolites in feces and 62 differential metabolites in urine, and the changes of the levels of these biomarkers showed that Semen Cuscutae had a good therapeutic effect on Kidney yang deficiency syndrome. Through the analysis of the channel, the metabolite changes mainly affected the steroid hormone biosynthesis, arachidonic acid metabolism, primary bile acid biosynthesis, sheath lipid metabolism and biosynthesis of tyrosine, phenylalanine metabolism, retinol metabolism,taurine and hypotaurine metabolism, lysine degradation and vitamin B6 metabolism, tryptophan metabolism, terpenoid backbone biosynthesis and starch and sucrose metabolism. Therefore, the results suggested that Semen Cuscutae could exert a good therapeutic effect by reversing the levels of some biomarkers.