Journal of Traditional Chinese Medicine ›› 2026, Vol. 46 ›› Issue (5): 1061-1071.DOI: 10.19852/j.cnki.jtcm.2026.05.001
• Original Articles • Previous Articles Next Articles
Sweated and non-sweated Danshen (Radix Salviae Miltiorrhizae) promoting macrophage M2 polarization through Notch1/Jagged1/ Hes1 pathway: a comparative research
SHAN Xiaoxiao1, LI Ruoqing1, HONG Bangzhen1, GU Liangliang2, CHENG Nuo2, WANG Guokai2, YU Nianjun3, WU Peiyun3, PENG Daiyin3, ZHANG Caiyun2, CHEN Weidong1(
), WANG Lei1()
- 1
School of Pharmacy, Anhui University of Chinese Medicine; Anhui Genuine Chinese Medicinal Materials Quality Improvement Collaborative Innovation Center; Institute of Pharmacokinetics, Anhui University of Chinese Medicine; Key Laboratory of Xin’an Medicine, Ministry of Education, Anhui University of Chinese Medicine Hefei 230012, China
2School of Pharmacy, Anhui University of Chinese Medicine; Anhui Genuine Chinese Medicinal Materials Quality Improvement Collaborative Innovation Center; Institute of Pharmacokinetics, Anhui University of Chinese Medicine Hefei 230012, China
3School of Pharmacy, Anhui University of Chinese Medicine ;Anhui Genuine Chinese Medicinal Materials Quality Improvement Collaborative Innovation Center Hefei 230012, China
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Received:2024-11-21Accepted:2025-06-11Online:2026-10-15Published:2026-10-09 -
Contact:CHEN Weidong, School of Pharmacy, Anhui University of Chinese Medicine, Hefei 230012, China. wdchen@ahtcm.edu.cn,Telephone: +86-15156091053; WANG Lei, School of Pharmacy, Anhui University of Chinese Medicine, Hefei 230012, China. wanglei@ahtcm.edu.cn,Telephone: +86-15805513357 -
About author:SHAN Xiaoxiao and LI Ruoqing are co-first authors and contributed equally to this work
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Supported by:The present study was supported by the National Natural Science Foundation of China: Biomimetic Construction and Performance Evaluation of Sodium Alginat-Zein/Nanohydroxyapatite Composite Scaffolds(51303006);Anhui Province Key Research and Development Program Project: Research on Key Technologies for Quality Improvement and Quality Evaluation of Anhui Genuine Medicinal Material “Gucheng Danshen”; Research on Key Technologies and Industrialization of the Classical Famous Prescription “Kaixin San”(1704a0802145);Anhui Province Key Research and Development Program Project: Research on Key Technologies for Quality Improvement and Quality Evaluation of Anhui Genuine Medicinal Material “Gucheng Danshen”; Research on Key Technologies and Industrialization of the Classical Famous Prescription “Kaixin San”(202203a07020031);University Natural Science Research Project of Anhui Province: Mechanistic Study on the “Guiding Drugs Upward” Effect of Acorus Tatarinowii in the Anti-Alzheimer’s Disease Classical Famous Prescription Kaixin San through Transporter Regulation and Competitive Protein Binding; Mechanistic Study of the Traditional Chinese Medicine Principle of “Removing Blood Stasis and Eliminating Masses”: Salvia Miltiorrhiza Suppresses Epithelial-Mesenchymal Transition and Lung Metastasis of Triple-negative Breast Cancer Through C-C Motif Chemokine Ligand 2/Colony Stimulating Factor 3-mediated Macrophage Polarization; Construction of a Borneol-metformin Dual-probe Fluorescent Nanoemulsion and Investigation of the Mechanisms Underlying Their Mutual Permeation Enhancement and Brain-targeted Drug Delivery(2408085MH229);University Natural Science Research Project of Anhui Province: Mechanistic Study on the “Guiding Drugs Upward” Effect of Acorus Tatarinowii in the Anti-Alzheimer’s Disease Classical Famous Prescription Kaixin San through Transporter Regulation and Competitive Protein Binding; Mechanistic Study of the Traditional Chinese Medicine Principle of “Removing Blood Stasis and Eliminating Masses”: Salvia Miltiorrhiza Suppresses Epithelial-Mesenchymal Transition and Lung Metastasis of Triple-negative Breast Cancer Through C-C Motif Chemokine Ligand 2/Colony Stimulating Factor 3-mediated Macrophage Polarization; Construction of a Borneol-metformin Dual-probe Fluorescent Nanoemulsion and Investigation of the Mechanisms Underlying Their Mutual Permeation Enhancement and Brain-targeted Drug Delivery(KJ2021A0586);University Natural Science Research Project of Anhui Province: Mechanistic Study on the “Guiding Drugs Upward” Effect of Acorus Tatarinowii in the Anti-Alzheimer’s Disease Classical Famous Prescription Kaixin San through Transporter Regulation and Competitive Protein Binding; Mechanistic Study of the Traditional Chinese Medicine Principle of “Removing Blood Stasis and Eliminating Masses”: Salvia Miltiorrhiza Suppresses Epithelial-Mesenchymal Transition and Lung Metastasis of Triple-negative Breast Cancer Through C-C Motif Chemokine Ligand 2/Colony Stimulating Factor 3-mediated Macrophage Polarization; Construction of a Borneol-metformin Dual-probe Fluorescent Nanoemulsion and Investigation of the Mechanisms Underlying Their Mutual Permeation Enhancement and Brain-targeted Drug Delivery(KJ2021ZD0065);Young Talents Promotion Project of China Association of Traditional Chinese Medicine(CACM-2022-QNRC2-B03)
Cite this article
SHAN Xiaoxiao, LI Ruoqing, HONG Bangzhen, GU Liangliang, CHENG Nuo, WANG Guokai, YU Nianjun, WU Peiyun, PENG Daiyin, ZHANG Caiyun, CHEN Weidong, WANG Lei. Sweated and non-sweated Danshen (Radix Salviae Miltiorrhizae) promoting macrophage M2 polarization through Notch1/Jagged1/ Hes1 pathway: a comparative research[J]. Journal of Traditional Chinese Medicine, 2026, 46(5): 1061-1071.
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Figure 1 Effect of SNSE on the expression of CD86 and CD206 A: representative flow-cytometry plots showing CD86 and CD206 expression; A1: negative control (unstained); A2: untreated control; A3: LPS group; A4: IL-4 group; A5: LPS + NST group; A6: LPS + SST group; A7: LPS + NSPA group; A8: LPS + SSPA group; B: percentage of M1 (CD86-positive) macrophages; C: percentage of M2 (CD206-positive) macrophages. Cells were treated for 24 h under the following conditions: control group, untreated cells; LPS group, lipopolysaccharide (2 μg/mL); IL-4 group, interleukin-4 (1 μg/mL); LPS + NST group, LPS (2 μg/mL) combined with non-sweated Danshen tanshinone extract (NST, 0.01 mg/mL); LPS + SST group, LPS (2 μg/mL) combined with sweated Danshen tanshinone extract (SST, 0.01 mg/mL); LPS + NSPA group, LPS (2 μg/mL) combined with non-sweated Danshen phenolic acid extract (NSPA, 0.1 mg/mL); LPS + SSPA group, LPS (2 μg/mL) combined with sweated Danshen phenolic acid extract (SSPA, 0.1 mg/mL). SNSE: sweated and non-sweated Danshen (Radix Salviae Miltiorrhizae) extracts; LPS: lipopolysaccharide; IL-4: interleukin-4; NST: non-sweated Danshen tanshinone extract; SST: sweated Danshen tanshinone extract; NSPA: non-sweated Danshen phenolic acid extract; SSPA: sweated Danshen phenolic acid extract; CD86: cluster of differentiation 86; CD206: cluster of differentiation 206; M1: classically activated macrophages; M2: alternatively activated macrophages; ANOVA: analysis of variance. Data are presented as mean ± standard deviation (n = 3). Statistical comparisons were performed by one-way ANOVA followed by Tukey's multiple-comparisons test. aP < 0.0001, dP < 0.01, versus the untreated control; bP < 0.0001, versus the LPS group; cP < 0.001, eP < 0.05, for SST versus NST.
Figure 1 Effect of SNSE on the expression of CD86 and CD206 A: representative flow-cytometry plots showing CD86 and CD206 expression; A1: negative control (unstained); A2: untreated control; A3: LPS group; A4: IL-4 group; A5: LPS + NST group; A6: LPS + SST group; A7: LPS + NSPA group; A8: LPS + SSPA group; B: percentage of M1 (CD86-positive) macrophages; C: percentage of M2 (CD206-positive) macrophages. Cells were treated for 24 h under the following conditions: control group, untreated cells; LPS group, lipopolysaccharide (2 μg/mL); IL-4 group, interleukin-4 (1 μg/mL); LPS + NST group, LPS (2 μg/mL) combined with non-sweated Danshen tanshinone extract (NST, 0.01 mg/mL); LPS + SST group, LPS (2 μg/mL) combined with sweated Danshen tanshinone extract (SST, 0.01 mg/mL); LPS + NSPA group, LPS (2 μg/mL) combined with non-sweated Danshen phenolic acid extract (NSPA, 0.1 mg/mL); LPS + SSPA group, LPS (2 μg/mL) combined with sweated Danshen phenolic acid extract (SSPA, 0.1 mg/mL). SNSE: sweated and non-sweated Danshen (Radix Salviae Miltiorrhizae) extracts; LPS: lipopolysaccharide; IL-4: interleukin-4; NST: non-sweated Danshen tanshinone extract; SST: sweated Danshen tanshinone extract; NSPA: non-sweated Danshen phenolic acid extract; SSPA: sweated Danshen phenolic acid extract; CD86: cluster of differentiation 86; CD206: cluster of differentiation 206; M1: classically activated macrophages; M2: alternatively activated macrophages; ANOVA: analysis of variance. Data are presented as mean ± standard deviation (n = 3). Statistical comparisons were performed by one-way ANOVA followed by Tukey's multiple-comparisons test. aP < 0.0001, dP < 0.01, versus the untreated control; bP < 0.0001, versus the LPS group; cP < 0.001, eP < 0.05, for SST versus NST.
Figure 2 Effect of SNSE on the mRNA expression of IL-6, IL-10, IL-12 and TNF-α in macrophages A: IL-6 mRNA expression after NST/SST treatment; B: IL-10 mRNA expression after NST/SST treatment; C: IL-12 mRNA expression after NST/SST treatment; D: TNF-α mRNA expression after NST/SST treatment; E: IL-6 mRNA expression after NSPA/SSPA treatment; F: IL-10 mRNA expression after NSPA/SSPA treatment; G: IL-12 mRNA expression after NSPA/SSPA treatment; H: TNF-α mRNA expression after NSPA/SSPA treatment. Cells were treated for 24 h. LPS was used at 2 μg/mL; NST and SST were used at 0.005, 0.01, and 0.02 mg/mL; NSPA and SSPA were used at 0.05, 0.1, and 0.2 mg/mL. SNSE: sweated and non-sweated Danshen (Radix Salviae Miltiorrhizae) extracts; LPS: lipopolysaccharide; NST: non-sweated Danshen tanshinone extract; SST: sweated Danshen tanshinone extract; NSPA: non-sweated Danshen phenolic acid extract; SSPA: sweated Danshen phenolic acid extract; mRNA: messenger RNA; IL-6: interleukin-6; IL-10: interleukin-10; IL-12: interleukin-12; TNF-α: tumor necrosis factor-α; ANOVA: analysis of variance. Data are presented as mean ± standard deviation (n = 6). Statistical comparisons were performed by one-way ANOVA followed by Tukey's multiple-comparisons test. aP < 0.01 versus the untreated control; bP < 0.01, dP < 0.05, versus the LPS group; cP < 0.05 for the sweated extract versus the corresponding non-sweated extract at the same concentration.
Figure 2 Effect of SNSE on the mRNA expression of IL-6, IL-10, IL-12 and TNF-α in macrophages A: IL-6 mRNA expression after NST/SST treatment; B: IL-10 mRNA expression after NST/SST treatment; C: IL-12 mRNA expression after NST/SST treatment; D: TNF-α mRNA expression after NST/SST treatment; E: IL-6 mRNA expression after NSPA/SSPA treatment; F: IL-10 mRNA expression after NSPA/SSPA treatment; G: IL-12 mRNA expression after NSPA/SSPA treatment; H: TNF-α mRNA expression after NSPA/SSPA treatment. Cells were treated for 24 h. LPS was used at 2 μg/mL; NST and SST were used at 0.005, 0.01, and 0.02 mg/mL; NSPA and SSPA were used at 0.05, 0.1, and 0.2 mg/mL. SNSE: sweated and non-sweated Danshen (Radix Salviae Miltiorrhizae) extracts; LPS: lipopolysaccharide; NST: non-sweated Danshen tanshinone extract; SST: sweated Danshen tanshinone extract; NSPA: non-sweated Danshen phenolic acid extract; SSPA: sweated Danshen phenolic acid extract; mRNA: messenger RNA; IL-6: interleukin-6; IL-10: interleukin-10; IL-12: interleukin-12; TNF-α: tumor necrosis factor-α; ANOVA: analysis of variance. Data are presented as mean ± standard deviation (n = 6). Statistical comparisons were performed by one-way ANOVA followed by Tukey's multiple-comparisons test. aP < 0.01 versus the untreated control; bP < 0.01, dP < 0.05, versus the LPS group; cP < 0.05 for the sweated extract versus the corresponding non-sweated extract at the same concentration.
Figure 3 Effect of SNSE on the concentration of IL-6, IL-10, IL-12 and TNF-α in macrophages A: IL-6 concentration after NST/SST treatment; B: IL-10 concentration after NST/SST treatment; C: IL-12 concentration after NST/SST treatment; D: TNF-α concentration after NST/SST treatment; E: IL-6 concentration after NSPA/SSPA treatment; F: IL-10 concentration after NSPA/SSPA treatment; G: IL-12 concentration after NSPA/SSPA treatment; H: TNF-α concentration after NSPA/SSPA treatment. Cytokine concentrations were determined by ELISA. Cells were treated for 24 h. LPS was used at 2 μg/mL; NST and SST were used at 0.005, 0.01, and 0.02 mg/mL; NSPA and SSPA were used at 0.05, 0.1, and 0.2 mg/mL. SNSE: sweated and non-sweated Danshen (Radix Salviae Miltiorrhizae) extracts; LPS: lipopolysaccharide; NST: non-sweated Danshen tanshinone extract; SST: sweated Danshen tanshinone extract; NSPA: non-sweated Danshen phenolic acid extract; SSPA: sweated Danshen phenolic acid extract; IL-6: interleukin-6; IL-10: interleukin-10; IL-12: interleukin-12; TNF-α: tumor necrosis factor-α; ELISA: enzyme-linked immunosorbent assay; ANOVA: analysis of variance. Data are presented as mean ± standard deviation (n = 6). Statistical comparisons were performed by one-way ANOVA followed by Tukey's multiple-comparisons test. aP < 0.05, eP < 0.01, versus the untreated control; bP < 0.05, cP < 0.01 versus the LPS group; dP < 0.05 for the sweated extract versus the corresponding non-sweated extract at the same concentration; versus the untreated control.
Figure 3 Effect of SNSE on the concentration of IL-6, IL-10, IL-12 and TNF-α in macrophages A: IL-6 concentration after NST/SST treatment; B: IL-10 concentration after NST/SST treatment; C: IL-12 concentration after NST/SST treatment; D: TNF-α concentration after NST/SST treatment; E: IL-6 concentration after NSPA/SSPA treatment; F: IL-10 concentration after NSPA/SSPA treatment; G: IL-12 concentration after NSPA/SSPA treatment; H: TNF-α concentration after NSPA/SSPA treatment. Cytokine concentrations were determined by ELISA. Cells were treated for 24 h. LPS was used at 2 μg/mL; NST and SST were used at 0.005, 0.01, and 0.02 mg/mL; NSPA and SSPA were used at 0.05, 0.1, and 0.2 mg/mL. SNSE: sweated and non-sweated Danshen (Radix Salviae Miltiorrhizae) extracts; LPS: lipopolysaccharide; NST: non-sweated Danshen tanshinone extract; SST: sweated Danshen tanshinone extract; NSPA: non-sweated Danshen phenolic acid extract; SSPA: sweated Danshen phenolic acid extract; IL-6: interleukin-6; IL-10: interleukin-10; IL-12: interleukin-12; TNF-α: tumor necrosis factor-α; ELISA: enzyme-linked immunosorbent assay; ANOVA: analysis of variance. Data are presented as mean ± standard deviation (n = 6). Statistical comparisons were performed by one-way ANOVA followed by Tukey's multiple-comparisons test. aP < 0.05, eP < 0.01, versus the untreated control; bP < 0.05, cP < 0.01 versus the LPS group; dP < 0.05 for the sweated extract versus the corresponding non-sweated extract at the same concentration; versus the untreated control.
Figure 4 Effects of NST, SST, NSPA, and SSPA on the protein expression of the Notch1/Jagged1/Hes1 signaling pathway in LPS-stimulated macrophages A: representative Western blots of Notch1, Jagged1, Hes1, and β-actin after NST/SST treatment; B: Notch1/β-actin after NST/SST treatment; C: Jagged1/β-actin after NST/SST treatment; D: Hes1/β-actin after NST/SST treatment; E: representative Western blots of Notch1, Jagged1, Hes1, and β-actin after NSPA/SSPA treatment; F: Notch1/β-actin after NSPA/SSPA treatment; G: Jagged1/β-actin after NSPA/SSPA treatment; H: Hes1/β-actin after NSPA/SSPA treatment. Molecular-mass values are aligned in a separate column, with one shared kDa label below the values. Cells were treated for 24 h. LPS was used at 2 μg/mL; NST and SST were used at 5, 10, and 20 μg/mL; NSPA and SSPA were used at 50, 100, and 200 μg/mL. SNSE: sweated and non-sweated Danshen (Radix Salviae Miltiorrhizae) extracts; LPS: lipopolysaccharide; NST: non-sweated Danshen tanshinone extract; SST: sweated Danshen tanshinone extract; NSPA: non-sweated Danshen phenolic acid extract; SSPA: sweated Danshen phenolic acid extract; kDa: kilodalton; ANOVA: analysis of variance. Data are presented as mean ± standard deviation (n = 3). Statistical comparisons were performed by one-way ANOVA followed by Tukey's multiple-comparisons test. aP < 0.05, eP < 0.01, versus the untreated control; bP < 0.05, cP < 0.01 versus the LPS group; dP < 0.05 for SST versus NST at the same concentration.
Figure 4 Effects of NST, SST, NSPA, and SSPA on the protein expression of the Notch1/Jagged1/Hes1 signaling pathway in LPS-stimulated macrophages A: representative Western blots of Notch1, Jagged1, Hes1, and β-actin after NST/SST treatment; B: Notch1/β-actin after NST/SST treatment; C: Jagged1/β-actin after NST/SST treatment; D: Hes1/β-actin after NST/SST treatment; E: representative Western blots of Notch1, Jagged1, Hes1, and β-actin after NSPA/SSPA treatment; F: Notch1/β-actin after NSPA/SSPA treatment; G: Jagged1/β-actin after NSPA/SSPA treatment; H: Hes1/β-actin after NSPA/SSPA treatment. Molecular-mass values are aligned in a separate column, with one shared kDa label below the values. Cells were treated for 24 h. LPS was used at 2 μg/mL; NST and SST were used at 5, 10, and 20 μg/mL; NSPA and SSPA were used at 50, 100, and 200 μg/mL. SNSE: sweated and non-sweated Danshen (Radix Salviae Miltiorrhizae) extracts; LPS: lipopolysaccharide; NST: non-sweated Danshen tanshinone extract; SST: sweated Danshen tanshinone extract; NSPA: non-sweated Danshen phenolic acid extract; SSPA: sweated Danshen phenolic acid extract; kDa: kilodalton; ANOVA: analysis of variance. Data are presented as mean ± standard deviation (n = 3). Statistical comparisons were performed by one-way ANOVA followed by Tukey's multiple-comparisons test. aP < 0.05, eP < 0.01, versus the untreated control; bP < 0.05, cP < 0.01 versus the LPS group; dP < 0.05 for SST versus NST at the same concentration.
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