Journal of Traditional Chinese Medicine ›› 2026, Vol. 46 ›› Issue (5): 1117-1126.DOI: 10.19852/j.cnki.jtcm.2026.05.006
• Original Articles • Previous Articles Next Articles
XU Hui1,2, QI Li3, QIAN Yutong1, LIU Chang1, LI Zhenchen3, MENG Qi3, LIU Shimin1, JIAO Danli1(
), ZHAO Chen1(
)
Received:2025-05-29
Accepted:2025-11-05
Online:2026-10-15
Published:2026-10-09
Contact:
Prof. ZHAO Chen, School of Acupuncture-moxibustion and Tuina, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China. ztzhaochen@126.com,Telephone: +86-13764276086; Prof. JIAO Danli, School of Acupuncture-moxibustion and Tuina, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China. 937095417@qq.com,Telephone: +86-18616124659
Supported by:XU Hui, QI Li, QIAN Yutong, LIU Chang, LI Zhenchen, MENG Qi, LIU Shimin, JIAO Danli, ZHAO Chen. Grain-sized moxibustion attenuates renal dysfunction and inflammation in naturally aging mice via the phosphatidylinositol 3-kinase/protein kinase B/nuclear factor-κB pathway[J]. Journal of Traditional Chinese Medicine, 2026, 46(5): 1117-1126.
Figure 1 GS-Moxi decreases age-associated renal dysfunction and histopathological morphology A: schematic diagram of the overall animal experimental design; B: representative PAS staining micrographs of mouse renal tissues (scale bar = 50 μm); B1: Young group; B2: Old group; B3: GS-Moxi group; B4: semiquantitative statistical scoring of renal tubular injury; C: representative Masson’s trichrome staining micrographs of mouse renal tissues (scale bar = 50 μm); C1: Young group; C2: Old group; C3: GS-Moxi group; C4: semiquantitative statistical scoring of renal interstitial fibrosis; D: relative mRNA expression of TGF-β1 detected by qPCR; E1: SCr concentration; E2: BUN concentration. Young group: untreated young mice without acupoint stimulation; Old group: naturally aged mice with spontaneous senescent renal injury; GS-Moxi group: aged mice treated with grain-sized moxibustion at Zusanli (ST36) acupoint for 12 consecutive weeks. GS-Moxi: grain-sized moxibustion; PAS: periodic acid-Schiff; qPCR: quantitative real-time polymerase chain reaction; TGF-β1: transforming growth factor beta 1; SCr: serum creatinine; BUN: blood urea nitrogen. Statistical analyses were measured by one-way analysis, and least significance difference test was performed for inter-group comparisons. All data were presented as mean ± standard deviation (n = 6). Compared with the young group, aP < 0.01, cP < 0.05; compared with the old group, bP < 0.01, dP < 0.05.
Figure 2 GS-Moxi attenuates aging-associated inflammation and oxidative stress A: representative images and quantitative analysis of SA‐β‐gal activity in renal sections (scale bar, 50 μm); A1: Young group; A2: Old group; A3: GS-Moxi group; A4: the percentages of SA-β-gal positive cells (arrow: stained positive cells); B1: the protein expressions of P16, and P21 were measured by Western blot analysis with specific primary antibodies. β-actin protein level was used as a loading control; B2: the protein expressions of P16; B3: the protein expressions of P21; C: p16 and p21mRNA expression levels were measured by PCR analysis; D1: GS-Moxi decreased the level of MDA; D2: GS-Moxi increased the level of SOD; E1: expression of TNF-α by ELISA; E2: expression of IL‐1β by ELISA; E3: expression of IL‐6 by ELISA. Young group, untreated young mice without acupoint stimulation; Old group, naturally aged mice with spontaneous senescent renal injury; GS-Moxi group, aged mice treated with grain-sized moxibustion at ST36 acupoint for 12 consecutive weeks. GS-Moxi: grain-sized moxibustion; SA-β-gal: senescence-associated β-galactosidase; qPCR: quantitative real-time polymerase chain reaction; MDA: malondialdehyde; SOD: superoxide dismutase; TNF-α: tumor necrosis factor alpha; IL-1β: interleukin 1 beta; IL-6: interleukin 6; ELISA: enzyme-linked immunosorbent assay. All quantitative data were presented as mean±standard deviation (n = 3 or n = 6 per group). One-way analysis of variance combined with Tukey’s multiple comparison post-hoc test was adopted to evaluate intergroup differences across Young, Old, and GS-Moxi groups. Compared with the young group, aP < 0.01, cP < 0.05; compared with the old group, bP < 0.01, dP < 0.05.
Figure 3 RNA-seq results of GS-Moxi-treated aging-induced mice A: PCA analysis; B: GSEA showed the differentially expressed gene sets involved in immune response; B1: young group; B2: GS-Moxi groups; C: heatmap of different genes related immunity reaction after GS-Moxi intervention among three groups. Red areas represent high expression, and blue areas represent low expression; D: KEGG pathway enrichment of differential gene expression among three groups. The PI3K/Akt/NF-κB signaling pathway was marked in red. Young group: untreated young mice without acupoint stimulation; Old group: naturally aged mice with spontaneous senescent renal injury; GS-Moxi group: aged mice treated with grain-sized moxibustion at Zusanli (ST36) acupoint for 12 consecutive weeks. GS-Moxi: grain-sized moxibustion; PCA: principal component analysis; GSEA: gene set enrichment analysis; KEGG: kyoto encyclopedia of genes and genomes; PI3K: phosphatidylinositol 3-kinase; AKT: protein kinase B; NF-κB: nuclear factor kappa-B. Sgk1: serum/ glucocorticoid regulated kinase 1; Creb3l3: cAMP responsive element binding protein 3-like 3; Igkv6-14: immunoglobulin kappa variable 6-14; Igkv9-124: immunoglobulin kappa variable 9-124; Wfdc17: WAP four-disulfide core domain 17; Lcn2: lipocalin 2; Igkv4-53: immunoglobulin kappa variable 4-53; Socs3: suppressor of cytokine signaling 3; Cfi: complement factor I; Vnn1: vanin 1; Bdkrb2: bradykinin receptor B2; Grem1: gremlin 1; Endou: poly(U) specific endonuclease; Igkv4-57-1: immunoglobulin kappa variable 4-57-1; Igkv4-68: immunoglobulin kappa variable 4-68; Igkv4-61: immunoglobulin kappa variable 4-61; Tyro3: TYRO3 protein tyrosine kinase; Igkv5-48: immunoglobulin kappa variable 5-48.
Figure 4 GS-Moxi can prevent age-related renal injury by inhibiting PI3K/Akt signaling pathway A: representative micrographs of immunohistochemical staining (scale bar, 100 µm); A1: p-PI3K in young group; A2: p-PI3K in Old group; A3: p-PI3K in Gs-Moxi group; A4: p-AKT in young group; A5: p-AKT in Old group; A6: p-AKT in Gs-Moxi group; B: immunohistochemical results in renal tissues of mice in each group; B1: p-AKT; B2: p-PI3K; C: representative Western blot bands of PI3K, p-PI3K, Akt, p-Akt, NF-κB and p-NF-κB in renal tissues. β-actin was used as a loading control; D: Western blot analysis to detect the expression levels of proteins in renal tissues; D1: p-PI3K/PI3K; D2: p-AKT/AKT; D3: p-NF-κB/NF-κB. Young group: untreated young mice without acupoint stimulation; Old group: naturally aged mice with spontaneous senescent renal injury; GS-Moxi group: aged mice treated with grain-sized moxibustion at Zusanli (ST36) acupoint for 12 consecutive weeks. GS-Moxi: grain-sized moxibustion; IHC: immunohistochemistry; PI3K: phosphatidylinositol 3-kinase; p-PI3K: phosphorylated phosphatidylinositol 3-kinase; AKT: protein kinase B; p-AKT: phosphorylated protein kinase B; NF-κB: nuclear factor kappa-B; p-NF-κB: phosphorylated nuclear factor kappa-B. Statistical analyses were measured by one-way analysis, and least significance difference test was performed for inter-group comparisons. Data were presented as mean ± standard deviation (n = 3). Compared with the young group, aP < 0.01, cP < 0.05; compared with the old group, bP < 0.01.
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& China Academy of Chinese Medical Sciences
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