Journal: bioRxiv

Article Title: Functional segregation of body-brain signals in the area postrema

doi: 10.64898/2026.06.15.732473

Figure Lengend Snippet: a, GFRAL neuron response aligned to liquid food access or IP injection. Traces are mean +/- SEM. Colored traces are statistically significant responses after multiple comparison correction. b, CALCR neuron response aligned to liquid food access or IP injection. c, PRLHR neuron response aligned to liquid food access or IP injection. d, Left, PRLHR neuron responses to an injection of angiotensin after a subcutaneous injection of the angiotensin-receptor blocker losartan (losar) or vehicle (veh) (colors per graph on right). Right, z scores (0–40 min). e, Left, PRLHR neuron responses to a subcutaneous injection of the osmotic agent polyethylene glycol (PEG) which causes hypovolemia. Prior to PEG administration animals received subq losartan + IP captopril (inh) which inhibit endogenous angiotensin signaling, or vehicle (veh). Right, z scores (0–40 min). Data shows that PRLHR neurons respond to hypovolemia in an angiotensin-dependent manner. f, SLC6A2 neuron response aligned to liquid food access or IP injection. g, Boxplot showing z-score (0-40 min) for SLC6A2 neuron responses. Colored boxes are statistically significant. Statistical comparisons are relative to the baseline prior to injection/ingestion. *P<0.05, **P<0.01, ****P<0.0001

Article Snippet: For injection experiments, mice were injected with compounds at the following concentrations based on previously published reports: amylin rat, 20 μg/kg (MedChemExpress); salmon calcitonin, 150 μg/kg (MedChemExpress); ghrelin, 2 mg/kg (R&D Systems); [D-Ala2]-GIP, 374 μg/kg (Biotechne); LiCl, 168 mg/kg; Deoxynivalenol (DON), 2.5 mg/kg (Tocris); exendin-4, 100 μg/kg (Bachem), 3M NaCl; angiotensin-II, 2.0 mg/kg (MedChemExpress); cisplatin 7.5 mg/kg (MedChemExpress), GDF-15, 25 μg/kg (MedChemExpress); devazepide, 1 mg kg/1 (R&D Systems); Exendin(9-39), 1 mg/kg (MedChemExpress); ponsegromab, 10 mg/kg, (MedChemExpress), losartan, 100 mg/kg (MedChemExpress); captopril, 50 mg/kg (MedChemExpress); isoproterenol 100 mg/kg (MEdCHemExpress).

Techniques: Injection, Comparison

Journal: Chinese Medicine

Article Title: Zhen-Wu-Tang ameliorates uremic cardiomyopathy via targeting the kidney–heart inflammatory axis and suppressing CCL2/CCR2-mediated macrophage activation

doi: 10.1186/s13020-026-01376-2

Figure Lengend Snippet: ZWT effectively alleviated uremic renal dysfunction in mice. ZWT Effectively Alleviated Uremic Renal Dysfunction in Mice. A Schematic of UC model establishment and ZWT treatment. Mice were administered ZWT at doses of 5 g/kg/d and 10 g/kg/d, Captopril (9.75 mg/kg/d), or Finerenone (10 mg/kg/d) for 8 weeks, following 5/6 nephrectomy. The sham group only underwent laparotomy and suturing, without kidney tissue resection. B Hematoxylin–eosin (HE) and Masson staining of kidney sections, n = 6 mouse kidneys per group, scale bar = 50 μm. C Quantification of the fibrotic area in Masson staining. D Blood urea nitrogen (BUN) levels and ( E ) Creatinine levels in blood were measured by ELISA, n = 8 mice per group. Data in C–E were presented as mean ± SD, data were analyzed using one-way ANOVA with Tukey's test, with p ≤ 0.05 considered statistically significant. * p ≤ 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001

Article Snippet: Efficacy Assessment of ZWT: Mice were randomized into six groups: sham, model, ZWT low-dose (5 g/kg/d), ZWT high-dose (10 g/kg/d), Captopril (9.75 mg/kg/d; CAS: 62571‐86‐2, MCE, USA), and Finerenone (10 mg/kg/d; Kerendia, BAYER, BXK01J1).

Techniques: Staining, Enzyme-linked Immunosorbent Assay

Journal: Chinese Medicine

Article Title: Zhen-Wu-Tang ameliorates uremic cardiomyopathy via targeting the kidney–heart inflammatory axis and suppressing CCL2/CCR2-mediated macrophage activation

doi: 10.1186/s13020-026-01376-2

Figure Lengend Snippet: ZWT Improved UC in Mice. A Representative images of heart sections stained with HE, Masson’s trichrome, n = 6 mouse hearts per group, M-mode echocardiography from the mice 8 weeks after 5/6 nephrectomy with or without ZWT treatment, n = 8–10 mice per group, scale bar = 50 μm. B Quantification of cardiomyocyte size based on HE staining, n = 6 samples per group. C–H Statistical analysis of heart weight/body weight (HW/BW), ejection fraction (EF), fractional shortening (FS), cardiac output, left ventricular posterior wall thickness at end-systole (LVPW; s), and left ventricular posterior wall in at end-diastole (LVPW; d), n = 8 mice per group. Data in B–H were presented as mean ± SD, data were analyzed using one-way ANOVA with Tukey's test, with p ≤ 0.05 considered statistically significant. * p ≤ 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001

Article Snippet: Efficacy Assessment of ZWT: Mice were randomized into six groups: sham, model, ZWT low-dose (5 g/kg/d), ZWT high-dose (10 g/kg/d), Captopril (9.75 mg/kg/d; CAS: 62571‐86‐2, MCE, USA), and Finerenone (10 mg/kg/d; Kerendia, BAYER, BXK01J1).

Techniques: Staining

Journal: Chinese Medicine

Article Title: Zhen-Wu-Tang ameliorates uremic cardiomyopathy via targeting the kidney–heart inflammatory axis and suppressing CCL2/CCR2-mediated macrophage activation

doi: 10.1186/s13020-026-01376-2

Figure Lengend Snippet: ZWT Attenuates Circulating Inflammatory Cytokines and Chemokines. ZWT Attenuates Circulating Inflammatory Cytokines and Chemokines. A Principal Component Analysis (PCA) of plasma secreted proteomics data comparing the sham group (blue), the UCM group (red), and the UCM with ZWT group (green), n = 2 samples per group. B Box plots showing expression values of samples in each group, n = 2 samples per group. C A heatmap illustrating the variations of 92 secreted proteins in mouse blood across the three groups. The data were presented as mean value of each group, n = 2 samples per group. D–E Gene Ontology (GO) enrichment analysis of biological processes (BP) and molecular functions (MF) for the top 30 proteins with the most significant reversed trends after ZWT treatment. F Circulating CCL2 levels were detected by ELISA, n = 8–10. Serum IL-1β levels quantified by ELISA, n = 6 mice per group. H Systemic IL-6 levels were measured by ELISA, n = 8–10 mice per group. Boxplots of B data were displayed the median as the center line, with the box limits representing the upper and lower quartiles. Data in F–H were presented as mean ± SD, data were analyzed using one-way ANOVA with Tukey's test, with p ≤ 0.05 considered statistically significant. * p ≤ 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001

Article Snippet: Efficacy Assessment of ZWT: Mice were randomized into six groups: sham, model, ZWT low-dose (5 g/kg/d), ZWT high-dose (10 g/kg/d), Captopril (9.75 mg/kg/d; CAS: 62571‐86‐2, MCE, USA), and Finerenone (10 mg/kg/d; Kerendia, BAYER, BXK01J1).

Techniques: Clinical Proteomics, Expressing, Enzyme-linked Immunosorbent Assay

Journal: Chinese Medicine

Article Title: Zhen-Wu-Tang ameliorates uremic cardiomyopathy via targeting the kidney–heart inflammatory axis and suppressing CCL2/CCR2-mediated macrophage activation

doi: 10.1186/s13020-026-01376-2

Figure Lengend Snippet: Qualitative Analysis of Major Components in ZWT. Qualitative Analysis of Major Components in ZWT. A Comprehensive qualitative analysis of ZWT components by UHPLC-Q/TOF–MS. Total ion chromatogram display separated constituents in positive (upper panel) and negative (lower panel) ionization modes. B Chemical structures of the 45 major compounds identified in ZWT

Article Snippet: Efficacy Assessment of ZWT: Mice were randomized into six groups: sham, model, ZWT low-dose (5 g/kg/d), ZWT high-dose (10 g/kg/d), Captopril (9.75 mg/kg/d; CAS: 62571‐86‐2, MCE, USA), and Finerenone (10 mg/kg/d; Kerendia, BAYER, BXK01J1).

Techniques:

Journal: Chinese Medicine

Article Title: Zhen-Wu-Tang ameliorates uremic cardiomyopathy via targeting the kidney–heart inflammatory axis and suppressing CCL2/CCR2-mediated macrophage activation

doi: 10.1186/s13020-026-01376-2

Figure Lengend Snippet: Identification of Blood-Absorbed Active Components of ZWT. Identification of Blood-Absorbed Active Components of ZWT. A Baseline blood (50 μl) was collected via orbital sinus puncture in rats as blank control plasma. Rats were orally administered ZWT (7 g/kg, crude drug equivalent) and plasma samples were collected at 0, 5, 15, 30 min and 1, 2, 4, 8, 12, 24 h post-ZWT administration. Post-dose plasma samples were pooled and analyzed by UHPLC-Q/TOF–MS. Total ion chromatograms are shown in positive (upper panel) and negative (lower panel) ionization modes; red labels highlight the 6 major components with their names and retention times (RT). B Critical cleavage pathways of four systemically absorbed components (Benzoylaconitine, paeoniflorin, atractylenolide III, and 6-Shogaol), comparing prototype compounds with their corresponding metabolites identified through pharmacokinetic profiling

Article Snippet: Efficacy Assessment of ZWT: Mice were randomized into six groups: sham, model, ZWT low-dose (5 g/kg/d), ZWT high-dose (10 g/kg/d), Captopril (9.75 mg/kg/d; CAS: 62571‐86‐2, MCE, USA), and Finerenone (10 mg/kg/d; Kerendia, BAYER, BXK01J1).

Techniques: Control, Clinical Proteomics

Journal: Chinese Medicine

Article Title: Zhen-Wu-Tang ameliorates uremic cardiomyopathy via targeting the kidney–heart inflammatory axis and suppressing CCL2/CCR2-mediated macrophage activation

doi: 10.1186/s13020-026-01376-2

Figure Lengend Snippet: ZWT active components suppressed TNFα-induced chemokine release and cellular injury in HK-2 cells. A Left: Venn diagram illustrating the overlap between the chemically characterized and the blood-absorbed components of ZWT. Right: Experimental scheme for the co-treatment of HK-2 cells with TNFα 10 ng/mL and ZWT active constituents for 24 h. B–G RT-qPCR analysis of chemokine-related gene expression including benzoylaconine, benzoylmesaconine, and hypaconitine at (10 and 20 μM), atractylenolide III and paeoniflorin at (30 and 60 μM), and 6-shogaol at (10 and 20 μg/mL) (n = 5–6). Heatmaps represent the average expression levels. H Radar plot illustrating the modulation patterns of chemokines based on data from B – G . Efficacy was calculated as the ratio of TNFα + ZWT high-dose / TNFα, with proximity to the center indicating stronger normalization. I ELISA quantification of CCL2 secretion in HK-2 cell supernatants (n = 3). J–L Relative mRNA expression of CCL5, CCL3, and CCL2 in HK-2 cells treated with individual or combined monomers including benzoylaconine (10 μM), atractylenolide III (30 μM), and paeoniflorin (30 μM). M Representative images and quantification of intracellular ROS levels in HK-2 cells (n = 3). N Representative western blot images of Bcl-2, Bax, and Cleaved-Caspase3 (n = 3). O–Q Densitometric quantification of Bcl-2, Bax, and Cleaved-Caspase3 levels normalized to Gapdh. Data in I–M and O–Q were analyzed using one-way ANOVA with Tukey's test, with p ≤ 0.05 considered statistically significant. * p ≤ 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001

Article Snippet: Efficacy Assessment of ZWT: Mice were randomized into six groups: sham, model, ZWT low-dose (5 g/kg/d), ZWT high-dose (10 g/kg/d), Captopril (9.75 mg/kg/d; CAS: 62571‐86‐2, MCE, USA), and Finerenone (10 mg/kg/d; Kerendia, BAYER, BXK01J1).

Techniques: Quantitative RT-PCR, Gene Expression, Expressing, Enzyme-linked Immunosorbent Assay, Western Blot

Journal: Chinese Medicine

Article Title: Zhen-Wu-Tang ameliorates uremic cardiomyopathy via targeting the kidney–heart inflammatory axis and suppressing CCL2/CCR2-mediated macrophage activation

doi: 10.1186/s13020-026-01376-2

Figure Lengend Snippet: ZWT Mitigated Macrophage-Mediated Inflammatory Responses and Cardiomyocyte Hypertrophy. ZWT Mitigated Macrophage-Mediated Inflammatory Responses and Cardiomyocyte Hypertrophy. A Schematic of a three-stage cell culture model. HK-2 cells were pretreated with TNFα in the presence or absence of benzoylaconine (20 μM), paeoniflorin (60 μM) and atractylenolide III (60 μM) for 6 h. After TNFα washout, these ingredients were reapplied, and HK-2 equilibrated for 12 h before supernatant collected. Macrophages pre-stimulated with PMA (10 ng/mL, 24 h) were cultured with HK-2-conditioned supernatants for 24 h. Resultant macrophage supernatants were then used to treat AC16 cardiomyocytes for 24 h prior to functional assays. B Phase-contrast micrographs of THP-1 cells: (Left) untreated cells exhibiting a round, suspension morphology; (Right) cells treated with PMA, showing adherent growth and an irregular morphology. Scale bar = 100 μm. C–E ELISA quantified THP-1 macrophage differentiation markers, n = 6. Data in C–E were analyzed by unpaired t-test. F–G IL-1β and IL-6 levels in macrophage media after exposure to monomer-treated HK-2 supernatants, measured by ELISA, n = 6. H IL-6 secretion levels in macrophage culture media were measured after treatment with 10 μM CCR2 antagonist 4 (CCR2A), followed by administration of three monomeric compounds. n = 6 samples per group. I–K Relative mRNA levels of M1 markers ( iNOS , CD86 ) and M2 marker ( CD163 ) in THP1 cells, normalized to 18S rRNA, n = 3 samples per group. L–N Relative mRNA levels of NPPA , NPPB , and MYH7 in AC16 cells, normalized to GAPDH , n = 3 samples per group, Data in F–N were analyzed by one-way ANOVA with Tukey's test. Data in C–N were presented as mean ± SD, with p ≤ 0.05 considered statistically significant. * p ≤ 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001

Article Snippet: Efficacy Assessment of ZWT: Mice were randomized into six groups: sham, model, ZWT low-dose (5 g/kg/d), ZWT high-dose (10 g/kg/d), Captopril (9.75 mg/kg/d; CAS: 62571‐86‐2, MCE, USA), and Finerenone (10 mg/kg/d; Kerendia, BAYER, BXK01J1).

Techniques: Cell Culture, Functional Assay, Suspension, Enzyme-linked Immunosorbent Assay, Marker

Journal: Chinese Medicine

Article Title: Zhen-Wu-Tang ameliorates uremic cardiomyopathy via targeting the kidney–heart inflammatory axis and suppressing CCL2/CCR2-mediated macrophage activation

doi: 10.1186/s13020-026-01376-2

Figure Lengend Snippet: CCL2 Blockade Attenuates Cardiorenal Injury in Uremic Cardiomyopathy. Mice were subjected to 5/6 nephrectomy and treated with either ZWT (10 g/kg/day by intragastric administration) or an anti-CCL2 antibody (10 mg/kg every 10 days by intraperitoneal injection) for 8 weeks. A Circulating level of IL-1β was measured by ELISA; n = 6 per group. B Representative images of kidney sections stained with hematoxylin–eosin (HE) and Masson (scale bar = 50 μm; n = 3 mice per group). C , D Relative mRNA expression of M1 macrophage polarization markers in kidney tissue was determined by qPCR; n = 6 per group. E–H Relative mRNA levels of kidney injury, inflammation, and fibrosis markers were assessed by qPCR. I Representative images of HE-stained heart sections (n = 3 mice per group) and M-mode echocardiography (n = 6 mice per group). J Quantification of cardiomyocyte cross-sectional area from HE-stained sections (n = 3 mice per group). K–O Echocardiographic analysis included ejection fraction (EF), fractional shortening (FS), left ventricular posterior wall thickness at end‑systole (LVPW; s), left ventricular posterior wall thickness at end‑diastole (LVPW;d), and left ventricular mass (LV Mass); n = 6 mice per group. P , Q Relative mRNA expression of Nppa and Nppb in heart tissue normalized to 18S rRNA; n = 6 per group. Data were presented as mean ± SD and were analyzed by one‑way ANOVA with Tukey’s test. p ≤ 0.05 considered statistically significant. * p ≤ 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001

Article Snippet: Efficacy Assessment of ZWT: Mice were randomized into six groups: sham, model, ZWT low-dose (5 g/kg/d), ZWT high-dose (10 g/kg/d), Captopril (9.75 mg/kg/d; CAS: 62571‐86‐2, MCE, USA), and Finerenone (10 mg/kg/d; Kerendia, BAYER, BXK01J1).

Techniques: Injection, Enzyme-linked Immunosorbent Assay, Staining, Expressing

Journal: Chinese Medicine

Article Title: Zhen-Wu-Tang ameliorates uremic cardiomyopathy via targeting the kidney–heart inflammatory axis and suppressing CCL2/CCR2-mediated macrophage activation

doi: 10.1186/s13020-026-01376-2

Figure Lengend Snippet: ZWT Blocked Pathological Cross-Talk in UC. Proposed mechanisms of ZWT in mitigating UC and renal dysfunction. ZWT suppressed TNFα-induced chemokine release in renal tubular epithelial cells and mitigated CCR2 + macrophage recruitment. These effects led to reduced levels of inflammatory cytokines in the blood and ultimately alleviated cardiomyocyte hypertrophy

Article Snippet: Efficacy Assessment of ZWT: Mice were randomized into six groups: sham, model, ZWT low-dose (5 g/kg/d), ZWT high-dose (10 g/kg/d), Captopril (9.75 mg/kg/d; CAS: 62571‐86‐2, MCE, USA), and Finerenone (10 mg/kg/d; Kerendia, BAYER, BXK01J1).

Techniques: