Targeting the Gut-Kidney Axis in Diarrhea with Kidney-Yang Deficiency Syndrome: The Role of Sishen Pills in Regulating TMAO-Mediated Inflammatory Response.

BACKGROUND Sishen Pills (SSPs) are commonly used to treat diarrhea with kidney-yang deficiency syndrome. Trimethylamine-N-oxide (TMAO) is produced through the metabolism of gut microbiota and can participate in diarrhea in kidney-yang deficiency syndrome by mediating the "gut-kidney axis" to transmit inflammatory factors. This study combined network pharmacology with animal experiments to explore whether SSPs can treat diarrhea with kidney-yang deficiency syndrome by affecting the interaction between TMAO and gut microbiota. MATERIAL AND METHODS A mouse model of diarrhea with kidney-yang deficiency syndrome was constructed by using adenine and Folium sennae decoction, and SSP decoction was used for treatment. This study utilized network pharmacology to predict the potential mechanisms of SSPs in treating diarrhea with kidney-yang deficiency syndrome. 16S rRNA high-throughput sequencing was used to analyze gut mucosal microbial characteristics. ELISA was used to measure TMAO, NOD-like receptor thermal protein domain associated protein 3 (NLRP3), interleukin-1ß (IL-1ß), and transforming growth factor-ß1 (TGF-ß1) levels. We performed Masson and immunohistochemical (Occludin, ZO-1) staining of kidney and small intestinal tissues. The fluorescein diacetate (FDA) hydrolysis spectrophotometric method was used to assess the microbial activity in contents of the small intestine. RESULTS Network pharmacology analysis revealed that SSPs can modulate 108 target points involved in the development of diarrhea, including IL-1ß and TNF. The experimental results demonstrated that SSP decoction significantly improved the general behavioral profiles of the mice, and also reduced TMAO, NLRP3, IL-1ß, and TGF-ß1 levels (P<0.05). Correlation analysis revealed significant positive correlations between TMAO concentrations and NLRP3, IL-1ß and TGF-ß1 levels (P<0.05). Pathological analysis revealed improvements in renal fibrosis and increased expression of the Occludin and ZO-1 proteins in intestinal tissue. In the SSP group, there was a significant increase in microbial activity (P<0.001). According to the sequencing results, the characteristic bacteria of the SSP and NR groups included Succinatimonas hippei, uncultured Solirubrobacter sp., and Clostridium tyrobutyricum. Furthermore, TMAO, NLRP3, IL-1ß, and TGF-ß1 were significantly positively correlated (P<0.05) with Succinatimonas hippei and Clostridium tyrobutyricum. By modulating Firmicutes, Succinatimonas hippei, and Clostridium tyrobutyricum, SSP decoction lowers TMAO levels to alleviate diarrhea with kidney-yang deficiency syndrome. CONCLUSIONS TMAO likely plays a significant role in the "gut-kidney axis" of diarrhea with kidney-yang deficiency syndrome. By adjusting gut microbiota to reduce the inflammatory response that is transmitted through the "gut-kidney axis" as a result of elevated TMAO levels, SSP decoction can alleviate diarrhea with kidney-yang deficiency syndrome.

TMAO is involved in kidney-yang deficiency syndrome diarrhea by mediating the "gut-kidney axis".

Background: Trimethylamine-N-oxide (TMAO) is a harmful metabolite dependent on the intestinal microbiota and excreted through the kidneys. According to numerous investigations, rich circulation concentrations of TMAO have been linked to kidney and gastrointestinal disorders. Through the "gut-kidney axis" mediated by TMAO, this research attempted to clarify the microbiological causes of kidney-yang deficiency syndrome diarrhea. Methods: Adenine and Folium Sennae were used to create a mouse model of kidney-yang deficiency syndrome diarrhea. 16S rRNA sequencing was used to identify the traits of the intestinal mucosal microbiota. ELISA was used to assess TMAO, transforming growth factor-ß1 (TGF-ß1), interleukin-1ß (IL-1ß), and NOD-like receptor thermal protein domain associated protein 3 (NLRP3). Kidney tissue fibrosis was evaluated using Masson's trichrome staining, and immunohistochemical labeling was used to investigate the protein expression of occludin and Zonula Occludens-1(ZO-1) in small intestine tissue. Microbial activity was determined by using fluorescein diacetate (FDA) hydrolysis spectrophotometry. Results: TMAO showed a positive correlation with NLRP3, IL-1ß and TGF-ß1, all of which exhibited substantial increases (P < 0.05). Significant renal fibrosis and decreased ZO-1 and occludin expression in small intestine tissues were detected in the model group. The sequencing results revealed alterations in both α and ß diversities of small intestinal mucosal microbiota. Elevated TMAO concentrations were potentially associated with increasing Firmicutes/Bacteroidota (F/B) ratios, Streptococcus, Pseudomonas and unclassified Clostridia UCG 014, but with decreasing Rothia and RB41 abundances. Conclusion: This study establishes a link between intestinal microbiota dysbiosis and elevated TMAO concentrations. TMAO can activate inflammatory responses and cytokines, contributing to kidney-yang deficiency syndrome diarrhea via the "gut-kidney axis". Moreover, TMAO may coincide with disruptions in the intestinal barrier and renal fibrosis. Dysfunction of the "gut-kidney axis" further elevates TMAO levels, perpetuating a vicious cycle.

Dysfunction of cecal microbiota and CutC activity in mice mediating diarrhea with kidney-yang deficiency syndrome.

Objective: Previous studies have indicated that diarrhea with kidney-yang deficiency syndrome leads to a disorder of small intestine contents and mucosal microbiota. However, the relationship of TMA-lyase (CutC) activity and TMAO with diarrhea with kidney-yang deficiency syndrome remains unexplored. Therefore, this study explores the relationship between cecal microbiota and choline TMA-lyase (CutC) activity, as well as the correlation between trimethylamine oxide (TMAO), inflammatory index, and CutC activity. Method: Twenty SPF-grade male KM mice were randomly divided into the normal group (CN) and the diarrhea model group (CD). Diarrhea mouse models were established by adenine combined with Folium sennae administration. CutC activity, TMAO, interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α) levels were detected, and the cecal content microbiota was sequenced. Result: After 14 days, diarrhea occurred in the CD group. Compared with the CN group, there was no significant change in the activity of CutC in the small intestine of the CD group, while the activity of CutC in the cecum was significantly increased, and the levels of TMAO, IL-6, and TNF-α showed a significant increase. The Chao1 index, Observed_species index, Shannon index, and Simpson index all exhibited a decreasing trend. The main changes at the bacterial genus level were Alistipes, Enterorhabdus, Desulfovibrio, Bacteroides, Candidatus_Saccharimonas, and [Ruminococcus]_torques_group. The results of LEfSe analysis, random forest analysis and ROC curve analysis revealed Paludicola, Blautia, Negativibacillus, Paraprevotella, Harryflintia, Candidatus_Soleaferrea, Anaerotruncus, Oscillibacter, Colidextribacter, [Ruminococcus]_torques_group, and Bacteroides as characteristic bacteria in the CD group. Correlation analysis showed a significant negative correlation between cecal CutC activity and Ligilactobacillus, and a significant positive correlation with Negativibacillus and Paludicola. The level of TMAO was significantly positively correlated with CutC activity and IL-6. Conclusion: Diarrhea with kidney-yang deficiency syndrome significantly affects the physiological status, digestive enzyme activity, CutC activity, TMAO levels, and inflammatory response in mice. Additionally, there are changes in the composition and function of cecal microbiota, indicating an important impact of diarrhea with kidney-yang deficiency syndrome on the host intestinal microbiota balance. The occurrence of diarrhea with kidney-yang deficiency syndrome may be associated with dysbiosis of intestinal microbiota, increased CutC activity, elevated TMAO levels, and heightened inflammatory factor levels.

Lipid-lowering and antioxidant effects of Polygonatum fermented liquor: a study on intestinal microbiota and brain-gut axis in mice.

Introduction: This study aims to investigate the effects of Polygonatum fermented liquor (PFL) on improving lipid metabolism and oxidative stress in mice by regulating the gut microbiota. Methods: Forty SPF-grade male Kunming mice were randomly divided into four groups: normal control group (NC), general liquor group (GC), fresh Polygonatum fermented liquor group (FPC), and nine-steam-nine-bask Polygonatum fermented liquor group (NPC). Each group was administered with sterile water, general liquor, fresh Polygonatum fermented liquor, and nine-steam-nine-bask Polygonatum fermented liquor, respectively, by gavage. The mice's liver, brain tissue, serum, and intestinal contents were collected. The indicators of oxidative stress in the liver, four blood lipid indicators, gamma-aminobutyric acid (GABA), and brain-derived neurotrophic factor (BDNF) levels in the brain tissue were measured, liver hematoxylin and eosin (HE) staining was performed, and the gut microbiota in the small intestine were analyzed using 16S rRNA second-generation sequencing technology. Results: Compared with the NC group, the NPC group showed significantly increased liver glutathione peroxidase (GSH-Px) content in mice (p < 0.05), reduced number of lipid droplets in the liver cells, and increased GABA and BDNF content in the brain tissues. The NPC group regulated lipid metabolism by lowering low-density lipoprotein cholesterol (LDL-C) and increasing high-density lipoprotein cholesterol (HDL-C) content in the mouse serum. Gut microbiota analysis showed significant changes in the gut microbiota of mice in the FPC and NPC groups, with increased richness and species diversity. These two groups increased the abundance of beneficial bacteria such as Lactobacillus, unclassified Muribaculaceae, unclassified Bacilli, and uncultured Bacteroidales bacterium while reducing the abundance of harmful bacteria such as Candidatus Arthromitus, and Staphylococcus, with a particularly significant reduction in Staphylococcus (p < 0.05). It is speculated that the two types of PFL may exert lipid-lowering and antioxidant effects by modulating the abundance of these dominant bacteria. Further studies showed that various environmental factors are closely related to the dominant gut bacteria. Malondialdehyde (MDA) was significantly negatively correlated with Lactobacillus and unclassified Bacilli, superoxide dismutase (SOD) was significantly negatively correlated with Staphylococcus (p < 0.01) and significantly negatively correlated with Candidatus Arthromitus (p < 0.05), and HDL-C was significantly negatively correlated with Staphylococcus and Facklamia (p < 0.05). Discussion: The two types of PFL chosen in this study may exert lipid-lowering and antioxidant effects by modulating the composition and function of the gut microbiota, providing guidance for the industrial application of Polygonatum.

Risk Factor Analysis for Predicting the Onset of Rotator Cuff Calcific Tendinitis Based on Artificial Intelligence.

Background: Symptomatic rotator cuff calcific tendinitis (RCCT) is a common shoulder disorder, and approaches combined with artificial intelligence greatly facilitate the development of clinical practice. Current scarce knowledge of the onset suggests that clinicians may need to explore this disease thoroughly. Methods: Clinical data were retrospectively collected from subjects diagnosed with RCCT at our institution within the period 2008 to 2020. A standardized questionnaire related to shoulder symptoms was completed in all cases, and standardized radiographs of both shoulders were extracted using a human-computer interactive electronic medical system (EMS) to clarify the clinical diagnosis of symptomatic RCCT. Based on the exclusion of asymptomatic subjects, risk factors in the baseline characteristics significantly associated with the onset of symptomatic RCCT were assessed via stepwise logistic regression analysis. Results: Of the 1,967 consecutive subjects referred to our academic institution for shoulder discomfort, 237 were diagnosed with symptomatic RCCT (12.05%). The proportion of women and the prevalence of clinical comorbidities were significantly higher in the RCCT cohort than those in the non-RCCT cohort. Stepwise logistic regression analysis confirmed that female gender, hyperlipidemia, diabetes mellitus, and hypothyroidism were independent risk factors for the entire cohort. Stratified by gender, the study found a partial overlap of risk factors contributing to morbidity in men and women. Diagnosis of hyperlipidemia, diabetes mellitus, and hypothyroidism in male cases and diabetes mellitus in female cases were significantly associated with symptomatic RCCT. Conclusion: Independent predictors of symptomatic RCCT are female, hyperlipidemia, diabetes mellitus, and hypothyroidism. Men diagnosed with hyperlipidemia, diabetes mellitus, and hypothyroidism are at high risk for symptomatic RCCT, while more medical attention is required for women with diabetes mellitus. Artificial intelligence offers pioneering innovations in the diagnosis and treatment of musculoskeletal disorders, and careful assessment through individualized risk stratification can help predict onset and targeted early stage treatment.

Bufei Decoction Improves Lung-Qi Deficiency Syndrome of Chronic Obstructive Pulmonary Disease in Rats by Regulating the Balance of Th17/Treg Cells.

Bufei decoction (BFD) has been applied to treat chronic obstructive pulmonary disease (COPD) for centuries as a recognized traditional Chinese herbal formula. However, mechanisms of BFD on COPD are unclear. This study conducts an inquiry into the underlying mechanisms of the therapeutic effect of BFD on COPD. A COPD rat model with qi deficiency in lungs was established through induction using cigarette and sawdust smoking combined with intratracheal instillation of lipopolysaccharide following BFD treatment for 28 days. Changes in Th17/Treg cells of COPD rats with the syndrome of lung qi deficiency after BFD administration were verified using pulmonary function, ELISA, flow cytometry, histopathology, and Western blotting assays. The findings showed that BFD protected COPD rats from decreased lung function and lung injury. BFD administration reduced proinflammatory cytokines IL-6 and IL-17 secretion, promoted inhibitory cytokines IL-10 and TGF-ß secretion, decreased Th17/Treg cell ratio, markedly downregulated the Th17 cell transcription factor ROR-γt expression, and upregulated transcription factor Foxp3 expression in Treg cells. We speculate that lung tonic soup improved pulmonary qi deficiency in rats with COPD by regulating the balance of Th17/Treg cells.