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| b2-adrenergic receptors are important for the renal protection induced by vagus nerve stimulation (VNS) or ultrasound (US).
Source publication
The cholinergic anti-inflammatory pathway (CAP) links the nervous and immune systems and modulates innate and adaptive immunity. Activation of the CAP by vagus nerve stimulation exerts protective effects in a wide variety of clinical disorders including rheumatoid arthritis and Crohn's disease, and in murine models of acute kidney injury including...
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Context 1
... induction by LPS and its suppression by nicotine treatment, as observed previously in peritoneal macrophages, 3 were confirmed by enzyme-linked immunosorbent assay in peritoneal macrophages (Supplementary Figure S3) and RAW 264.7 cells (Figure 3a). Nicotine-induced TNF reduction was weakened when Hes1 expression was suppressed by small, interfering RNA (Figure 3b; Supplementary Figure S4 shows b a s i c r e s e a r c h T Inoue et al.: Hes1 in the cholinergic anti-inflammatory pathway small, interfering RNA efficiency). In addition, the induction of TNF expression by LPS was reduced by overexpression of Hes1 (Figure 3c). ...
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... 24 hours after transfection, the cells were used for further analysis. The knockdown efficiencies of Hes1 were validated by quantitative real-time polymerase chain reaction (CFX96, Bio-Rad) using the same primers described in Table S3 (Supplementary Figure S4). ...
Citations
... In addition, this study found that the functional nAChRs expressed on renal tubular epithelial cells may serve as a target for the cholinergic anti-inflammatory pathway. Other studies have indicated that stimulation of C1 neurons or pulsed ultrasound therapy can also alleviate renal IRI in mice by activating the cholinergic anti-inflammatory pathway [140][141][142]. However, α7nAChR antagonist treatment or α7nAChR gene knockout eliminated protective effects. ...
Ischemia-reperfusion injury (IRI) encompasses the damage resulting from the restoration of blood supply following tissue ischemia. This phenomenon commonly occurs in clinical scenarios such as hemorrhagic shock, severe trauma, organ transplantation, and thrombolytic therapy. Despite its prevalence, existing treatments exhibit limited efficacy against IRI. Vagus nerve stimulation (VNS) is a widely utilized technique for modulating the autonomic nervous system. Numerous studies have demonstrated that VNS significantly reduces IRI in various organs, including the heart, brain, and liver. This article reviews the pathological processes during IRI and summarizes the role and possible mechanisms of VNS in IRI of different organs. Furthermore, this review addresses the current challenges of VNS clinical applications, providing a novel perspective on IRI treatment.
... Moreover, a previous study demonstrated that intravenous adoptive transfer of peritoneal macrophages obtained from peritoneal lavage following vagus nerve stimulation conferred protection to the kidneys against ischemia-reperfusion injury. 42 These findings suggest that vagal signaling may exert a significant influence on peritoneal immunity and endow peritoneal macrophages with the capability to attenuate inflammation. Therefore, further functional analysis of how intraperitoneal cells behave might reveal noticeable differences in vagotomized mice. ...
The vagus nerve is the only pathway for transmitting parasympathetic signals between the brain and thoracoabdominal organs, thereby exhibiting anti-inflammatory functions through the cholinergic anti-inflammatory pathway. Despite often being resected during lymph node dissection in upper gastrointestinal cancer surgery, the impact of vagotomy on postoperative outcomes in gastric cancer patients remains unclear. Sub-diaphragmatic vagotomy was performed on C57BL/6 mice. Three weeks later, syngeneic murine gastric cancer cell line YTN16P was injected into the peritoneal cavity, and the number of peritoneal metastases (PM) on the mesentery and omentum compared with control mice. The phenotypes of immune cells in peritoneal lavage and omental milky spots one day after tumor inoculation were analyzed using flow cytometry and immunohistochemistry. Intraperitoneal transfer of 3 × 10⁵ YTN16P significantly increased the number of metastatic nodules on the mesentery in the vagotomy group compared to the control group. The omental metastasis grade was also significantly higher in the vagotomy group. Phenotypic analysis of immune cells in peritoneal lavage did not reveal significant differences after vagotomy. However, vagotomized mice exhibited a notable increase in milky spot area, with a higher presence of cytokeratin(+) tumor cells, F4/80(+) macrophages, and CD3(+) T cells. Vagus nerve signaling appears to regulate the immune response dynamics within milky spots against disseminated tumor cells and inhibits the development of PM. Preserving the vagus nerve may offer advantages in advanced gastric cancer surgery to reduce peritoneal recurrence.
... sepsis) (Borovikova et al. 2000;Tracey 2002;Huston et al. 2006;Rosas-Ballina et al. 2011;. Beyond inflammatory conditions, pFUS applied to the hepatoportal nerve plexus in the liver has been shown to restore glucose homeostasis in preclinical diabetic models Cotero et al. 2022) and applied to the spleen blocked ischemia/reperfusion injury (IRI) in the kidneys in preclinical IRI models (Gigliotti et al. 2013;Gigliotti et al. 2015;Inoue et al. 2019). ...
Background
Noninvasive ultrasound (US) has been used therapeutically for decades, with applications in tissue ablation, lithotripsy, and physical therapy. There is increasing evidence that low intensity US stimulation of organs can alter physiological and clinical outcomes for treatment of health disorders including rheumatoid arthritis and diabetes. One major translational challenge is designing portable and reliable US devices that can be used by patients in their homes, with automated features to detect rib location and aid in efficient transmission of energy to organs of interest. This feasibility study aimed to assess efficacy in rib bone detection without conventional imaging, using a single channel US pitch-catch technique integrated into an US therapy device to detect pulsed US reflections from ribs.
Methods
In 20 healthy volunteers, the location of the ribs and spleen were identified using a diagnostic US imaging system. Reflected ultrasound signals were recorded at five positions over the spleen and adjacent ribs using the therapy device. Signals were classified as between ribs (intercostal), partially over a rib, or fully over a rib using four models: threshold-based time domain classification, threshold-based frequency domain classification, logistic regression, and support vector machine (SVM).
Results
SVM performed best overall on the All Participants cohort with accuracy up to 96.25%. All models’ accuracies were improved by separating participants into two cohorts based on Body Mass Index (BMI) and re-fitting each model. After separation into Low BMI and High BMI cohorts, a simple time-thresholding approach achieved accuracies up to 100% and 93.75%, respectively.
Conclusion
These results demonstrate that US reflection signal classification can accurately provide low complexity, real-time automated onboard rib detection and user feedback to advance at-home therapeutic US delivery.
... Inoue et al. showed that VNS attenuated kidney injury in an ischemia/reperfusion injury-induced AKI model [60]. This renal protective effect required α7nAChR-positive splenocytes [61,62]. However, it is still unclear how the kidney receives VNS and the signals from splenic macrophages. ...
Acute kidney injury (AKI) is an emerging public health problem worldwide and is associated with high morbidity and mortality. The high mortality rate can be attributed to the lack of pharmacological therapies to prevent and treat AKI. Renal replacement therapy (RRT) plays a pivotal role in the treatment of patients with severe AKI. However, the mortality rate of patients with AKI requiring RRT exceeds 50%. Although studies on RRT for AKI have begun to resolve some of the associated problems, many issues remain to be addressed. Notably, the optimal timing of the initiation of RRT for AKI is still being debated. Recently, new therapeutic strategies for AKI have been developed. Angiotensin II and recombinant alkaline phosphatase treatment are expected to improve the clinical outcomes of patients with distributive and vasodilatory shock. Moreover, mitochondrial-targeted agents have been developed for the treatment of patients with AKI. This review is focused on the optimal timing of RRT for AKI and the new pharmacological interventions and therapies for AKI.
... Stimulation of the vagus nerve electrically [107][108][109][110], optogenetically [86,90,111], or by ultrasound [112,113] causes an anti-inflammatory effect in rodents and humans [114]. In fact, vagus nerve stimulation is considered a potential treatment for a wide variety of inflammatory conditions and injuries [115], including: sepsis [107]; postoperative ileus [109]; pancreatitis [86,116]; arthritis [114,117]; Crohn's disease [118,119]; and ischemia and reperfusion injury of the kidney [111,120,121] and heart [110,122,123]. ...
The vagus nerve vitally connects the brain and body to coordinate digestive, cardiorespiratory, and immune functions. Its efferent neurons, which project their axons from the brainstem to the viscera, are thought to comprise "functional units" - neuron populations dedicated to the control of specific vagal reflexes or organ functions. Previous research indicates that these functional units differ from one another anatomically, neurochemically, and physiologically but have yet to define their identity in an experimentally tractable way. However, recent work with genetic technology and single-cell genomics suggests that genetically distinct subtypes of neurons may be the functional units of the efferent vagus. Here we review how these approaches are revealing the organizational principles of the efferent vagus in unprecedented detail.
... In vitro experiments using antisense oligonucleotides on primary cultures of human macrophages identified the mediating role of alpha 7 nicotinic acetylcholine receptor (α7nAChR) 4 . VNS has been shown to ameliorate various inflammatory diseases, including sepsis 5,6 , rheumatoid arthritis 7,8 , gastrointestinal diseases (including Chron's disease and inflammatory bowel diseases) [9][10][11][12] , lung disease 13 , and acute kidney injury (AKI) [14][15][16] . ...
... Regarding the intracellular signaling pathways, experiments have shown that, after binding, α7nAChR inhibits nuclear translocation of nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) [30][31][32][33][34] , suppresses pyrin domaincontaining 3 (NLRP3) inflammasome activation 35 , and activates the Janus kinase 2 (JAK2)/signal transducer and activator of the transcription-3 (STAT3) pathway 36,37 . Recently, the hairy and enhancer of split-1 (Hes1), a transcriptional modulator, was also identified in macrophages 15 . Moreover, we previously reported that VNS ameliorates renal bilateral ischemia-reperfusion injury (IRI) in addition to systemic inflammation 14,15 . ...
... Recently, the hairy and enhancer of split-1 (Hes1), a transcriptional modulator, was also identified in macrophages 15 . Moreover, we previously reported that VNS ameliorates renal bilateral ischemia-reperfusion injury (IRI) in addition to systemic inflammation 14,15 . Ultrasound has also been shown to improve systemic inflammation, renal bilateral IRI kidney damage, and VNS by activating splenic CAP 38,39 . ...
Activation of the cholinergic anti-inflammatory pathway (CAP) via vagus nerve stimulation has been shown to improve acute kidney injury in rodent models. While alpha 7 nicotinic acetylcholine receptor (α7nAChR) positive macrophages are thought to play a crucial role in this pathway, their in vivo significance has not been fully understood. In this study, we used macrophage-specific α7nAChR-deficient mice to confirm the direct activation of α7nAChRs in macrophages. Our findings indicate that the administration of GTS-21, an α7nAChR-specific agonist, protects injured kidneys in wild-type mice but not in macrophage-specific α7nAChR-deficient mice. To investigate the signal changes or cell reconstructions induced by α7nAChR activation in splenocytes, we conducted single-cell RNA-sequencing of the spleen. Ligand-receptor analysis revealed an increase in macrophage-macrophage interactions. Using macrophage-derived cell lines, we demonstrated that GTS-21 increases cell contact, and that the contact between macrophages receiving α7nAChR signals leads to a reduction in TNF-α. Our results suggest that α7nAChR signaling increases macrophage-macrophage interactions in the spleen and has a protective effect on the kidneys.
... The efficacy of vagus nerve stimulation (VNS) in CAP activation is well established in animal disease models and has led to VNS-based pilot clinical trials for patients with Crohn's disease and rheumatoid arthritis [4]. Recently, CAP stimulation by electrical VNS or splenic pulsed ultrasound (pUS) has been found to successfully ameliorate experimental AKI following renal ischemiareperfusion injury (IRI) and endotoxemia [5][6][7]. We summarize the evidence for the role of CAP in preventing experimental AKI. ...
Inflammation is common in patients with acute kidney injury (AKI) and contributes to increased risk of morbidity and mortality. The central nervous system plays an important role in the immune and inflammatory pathways of AKI. In this review, we discuss the preclinical evidence for the neural pathways associated with neuromodulation in AKI and their clinical implications, as well as clinical trials that translate these observations into the clinical context. The ultimate goal of these trials is to design strategies using non-invasive approaches, such as splenic pulsed ultrasonography, to prevent or attenuate inflammatory conditions at the bedside, including AKI.
... Animal studies demonstrate that activation of CAP leads to the release of acetylcholine, which in turn exerts direct renoprotective effects on the kidney. [11][12][13][14][15] Indirect mechanisms by which CAP activation may favorably affect kidney function include reduction in inflammatory mediators 16,17 as well as regulation of heart rate and blood pressure via parasympathetic (vagal nerve) stimulation. 5,18,19 Cholinesterase inhibitors (ChEIs), namely donepezil, galantamine, and rivastigmine, are approved pharmacological therapies with the potential to offset cognitive decline in persons with Alzheimer's dementia (AD). ...
... 43,44 A series of animal studies have shown that the application of efferent vagus nerve stimulation and brainstem C1 neuron stimulation can reduce kidney damage and protect the kidney from ischemia reperfusion by activating the CAP through the splenic nerve. [11][12][13]15 In a rat model, activation of the CAP reduced chronic allograft nephropathy without any side effects for the recipient. 16 Another animal study showed that use of selective nicotinic acetylcholine receptor agonist may improve autonomic control, inhibit nuclear factor kB activation, and reduce renal Q11 , fibrosis and inflammatory response via CAP activation. ...
Preclinical evidence shows that activation of the cholinergic anti-inflammatory pathway (CAP) may have direct and indirect beneficial effects on the kidney. Cholinesterase inhibitors (ChEIs) are specific Alzheimer’s dementia (AD) therapies that block the action of cholinesterases and activate CAP. Here, we explored a plausible effect of ChEIs on slowing kidney function decline by comparing the risk of CKD progression among patients with newly diagnosed AD that initiated ChEI or not within 90 days. Using complete information of routine serum creatinine tests, we evaluated changes in estimated glomerular filtration rate (eGFR) and defined the outcome of chronic kidney disease (CKD) progression as the composite of an eGFR decline of over 30%, initiation of dialysis/transplant or death attributed to CKD. A secondary outcome was death. Inverse probability of treatment weighting Cox regression was used to estimate hazard ratios. Among 11, 898 patients, 6,803 started on ChEIs and 5,095 did not. Mean age was 80 years (64% women) and the mean eGFR was 68 ml/min/1.73m². During a median 3.0 years of follow-up, and compared to non-use, ChEI use was associated with 18% lower risk of CKD progression (1,231 events, adjusted hazard ratio 0.82; 95% confidence interval 0.71-0.96) and a 21% lower risk of death (0.79; 0.72-0.86). Results were consistent across subgroups, ChEI subclasses and after accounting for competing risks. Thus, in patients with AD undergoing routine care, use of ChEI (vs no-use) was associated with lower risk of CKD progression.
... transcription factors such as Hif(32), Irf1(33), Klf4(25), Hes1(34), and Klf6(26) (Figures 4D, E)involved in AKI progression, which may be important biomarkers for screening AKI progression. We also found PTC-cycling and part of PTC-new expressed proliferative markers such as Stmn1 and Pcna (Figure 4F). ...
Background
Understanding the acute kidney injury (AKI) microenvironment changes and the complex cellular interaction is essential to elucidate the mechanisms and develop new targeted therapies for AKI.
Methods
We employed unbiased single-cell RNA sequencing to systematically resolve the cellular atlas of kidney tissue samples from mice at 1, 2 and 3 days after ischemia-reperfusion AKI and healthy control. The single-cell transcriptome findings were validated using multiplex immunostaining, western blotting, and functional experiments.
Results
We constructed a systematic single-cell transcriptome atlas covering different AKI timepoints with immune cell infiltration increasing with AKI progression. Three new proximal tubule cells (PTCs) subtypes (PTC-S1-new/PTC-S2-new/PTC-S3-new) were identified, with upregulation of injury and repair-regulated signatures such as Sox9, Vcam1, Egr1, and Klf6 while with downregulation of metabolism. PTC-S1-new exhibited pro-inflammatory and pro-fibrotic signature compared to normal PTC, and trajectory analysis revealed that proliferating PTCs were the precursor cell of PTC-S1-new, and part of PTC-S1-new cells may turn into PTC-injured and then become fibrotic. Cellular interaction analysis revealed that PTC-S1-new and PTC-injured interacted closely with infiltrating immune cells through CXCL and TNF signaling pathways. Immunostaining validated that injured PTCs expressed a high level of TNFRSF1A and Kim-1, and functional experiments revealed that the exogenous addition of TNF-α promoted kidney inflammation, dramatic injury, and specific depletion of TNFRSF1A would abrogate the injury.
Conclusions
The single-cell profiling of AKI microenvironment provides new insight for the deep understanding of molecular changes of AKI, and elucidates the mechanisms and developing new targeted therapies for AKI.
... The preserved intestinal acetylcholine (4) in fish may enhance a7nAChR and had a mediating anti-inflammatory effect on immune cells, such as macrophages, neutrophils, and T cells (11)(12)(13)(14). When considering effectors, fish SBMIE showed typical allergic cytokine profiles (15), similar to IBD in humans. ...
Foodborne intestinal inflammation is a major health and welfare issue in aquaculture. To prevent enteritis, various additives have been incorporated into the fish diet. Considering anti-inflammatory immune regulation, an effective natural compound could potentially treat or prevent intestinal inflammation. Our previous study has revealed galantamine’s effect on soybean induced enteritis (SBMIE) and has highlighted the possible role of the cholinergic anti-inflammatory pathway in the fish gut. To further activate the intestinal cholinergic related anti-inflammatory function, α7nAchR signaling was considered. In this study, sinomenine, a typical agonist of α7nAChR in mammals, was tested to treat fish foodborne enteritis via its potential anti-inflammation effect using the zebrafish foodborne enteritis model. After sinomenine’s dietary inclusion, results suggested that there was an alleviation of intestinal inflammation at a pathological level. This outcome was demonstrated through the improved morphology of intestinal villi. At a molecular level, SN suppressed inflammatory cytokines’ expression (especially for tnf-α) and upregulated anti-inflammation-related functions (indicated by expression of il-10, il-22, and foxp3a). To systematically understand sinomenine’s intestinal effect on SBMIE, transcriptomic analysis was done on the SBMIE adult fish model. DEGs (sinomenine vs soybean meal groups) were enriched in GO terms related to the negative regulation of lymphocyte/leukocyte activation and alpha-beta T cell proliferation, as well as the regulation of lymphocyte migration. The KEGG pathways for glycolysis and insulin signaling indicated metabolic adjustments of α7nAchR mediated anti-inflammatory effect. To demonstrate the immune cells’ response, in the SBMIE larva model, inflammatory gatherings of neutrophils, macrophages, and lymphocytes caused by soybean meal could be relieved significantly with the inclusion of sinomenine. This was consistent within the sinomenine group as CD4⁺ or Foxp3⁺ lymphocytes were found with a higher proportion at the base of mucosal folds, which may suggest the Treg population. Echoing, the sinomenine group’s 16s sequencing result, there were fewer enteritis-related TM7, Sphingomonas and Shigella, but more Cetobacterium, which were related to glucose metabolism. Our findings indicate that sinomenine hydrochloride could be important in the prevention of fish foodborne enteritis at both immune and microbiota levels.
