Journal of Traditional Chinese Medicine ›› 2025, Vol. 45 ›› Issue (5): 963-969.DOI: 10.19852/j.cnki.jtcm.2025.05.003
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Shisiwei Jianzhong decoction (十四味建中汤) inhibits the adipogenic differentiation of bone marrow mesenchymal stem cells by downregulating nuclear factor of activated T cells, cytoplasmic 4 in non-severe aplastic anemia
WANG Jun, WANG Bo, ZHANG Yun, LIN Shengyun, WU Liqiang(
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- Department of Hematology, the First Affiliated Hospital of Zhejiang Chinese Medical University (Zhejiang Provincial Hospital of Chinese Medicine), Hangzhou 310006, China
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Received:2024-05-12Accepted:2024-12-23Online:2025-10-15Published:2025-09-15 -
Contact:WU Liqiang, Department of Hematology, the First Affiliated Hospital of Zhejiang Chinese Medical University (Zhejiang Provincial Hospital of Chinese Medicine), Hangzhou 310006, China. wuliqiang_2003@163.com, Tel: +86-15958158492 -
Supported by:Zhejiang Province Traditional Chinese Medicine Science and Technology Plan Project: Research on the Mechanism of Shisiwei Jianzhong Decoction in Treating Non-severe Aplastic Anemia with Kidney Yang Deficiency Based on the C Vactivated T Nuclear Factor (NFATc4)(2020ZB086);Zhejiang Province Traditional Chinese Medicine Science and Technology Youth Talent Plan Project: Study on the Academic thought and Clinical Application of Professor Zhou Yuhong in the Treatment of Chronic Aplastic Anemia(2021ZQ029)
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WANG Jun, WANG Bo, ZHANG Yun, LIN Shengyun, WU Liqiang. Shisiwei Jianzhong decoction (十四味建中汤) inhibits the adipogenic differentiation of bone marrow mesenchymal stem cells by downregulating nuclear factor of activated T cells, cytoplasmic 4 in non-severe aplastic anemia[J]. Journal of Traditional Chinese Medicine, 2025, 45(5): 963-969.
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Figure 1 SJD inhibited apoptosis of NSAA-derived BMSCs A: cell viability was measured by CCK-8 assay; n = 4; B: cell apoptosis was measured by flow cytometry. B1: control group; B2: 0.08 μg/mL group; B3: 0.8 μg/mL group; B4: 8 μg/mL group; B5: 80 μg/mL group; B6: quantitative analysis of apoptosis. Control group: basal medium; 0.08 μg/mL group: BMSCs were treated with 0.08 μg/mL SJD for 48 h; 0.8 μg/mL group: BMSCs were treated with 0.8 μg/mL SJD for 48 h; 8 μg/mL group: BMSCs were treated with 8 μg/mL SJD for 48 h; 80 μg/mL group: BMSCs were treated with 80 μg/mL SJD for 48 h;. SJD: Shisiwei Jianzhong decoction; NSAA: non-severe aplastic anemia; BMSCs: bone marrow mesenchymal stem cells; CCK-8: cell-counting kit-8. Statistical analysis was performed using one-way analysis of variance. Data are exhibited as mean ± standard deviation (n = 3). Compared with Control group, aP < 0.01 and bP < 0.05.
Figure 1 SJD inhibited apoptosis of NSAA-derived BMSCs A: cell viability was measured by CCK-8 assay; n = 4; B: cell apoptosis was measured by flow cytometry. B1: control group; B2: 0.08 μg/mL group; B3: 0.8 μg/mL group; B4: 8 μg/mL group; B5: 80 μg/mL group; B6: quantitative analysis of apoptosis. Control group: basal medium; 0.08 μg/mL group: BMSCs were treated with 0.08 μg/mL SJD for 48 h; 0.8 μg/mL group: BMSCs were treated with 0.8 μg/mL SJD for 48 h; 8 μg/mL group: BMSCs were treated with 8 μg/mL SJD for 48 h; 80 μg/mL group: BMSCs were treated with 80 μg/mL SJD for 48 h;. SJD: Shisiwei Jianzhong decoction; NSAA: non-severe aplastic anemia; BMSCs: bone marrow mesenchymal stem cells; CCK-8: cell-counting kit-8. Statistical analysis was performed using one-way analysis of variance. Data are exhibited as mean ± standard deviation (n = 3). Compared with Control group, aP < 0.01 and bP < 0.05.
Figure 2 SJD inhibited adipogenic differentiation of NSAA-derived BMSCs A: oil Red O staining of lipid droplets (× 40, Scale bar = 50 μm). A1: Control group; A2: 0.08 μg/mL group; A3: 0.8 μg/mL group; A4: 8 μg/mL group; A5: 80 μg/mL group. Control group: basal medium; 0.08 μg/mL group: BMSCs were treated with 0.08 μg/mL SJD for 48 h; 0.8 μg/mL group: BMSCs were treated with 0.8 μg/mL SJD for 48 h; 8 μg/mL group: BMSCs were treated with 8 μg/mL SJD for 48 h; 80 μg/mL group: BMSCs were treated with 80 μg/mL SJD for 48 h; B: relative mRNA levels of PPARG and FABP4 were detected by qRT-PCR. B1: quantitative analysis of PPARG levels; B2: quantitative analysis of FABP4 levels; C: the protein levels of PPARG and FABP4 were detected by Western blot. C1: protein images of PPARG and FABP4; C2: quantitative analysis of PPARG levels; C3: quantitative analysis of FABP4 levels. SJD: Shisiwei Jianzhong decoction; NSAA: non-severe aplastic anemia; BMSCs: bone marrow mesenchymal stem cells; PPARG: peroxisome proliferator-activated receptor gamma; FABP4: fatty acid-binding protein 4; qRT-PCR: quantitative real-time polymerase chain reaction. Statistical analysis was performed using one-way analysis of variance. Data are exhibited as mean ± standard deviation (n = 3). Compared with Control group, aP < 0.05 and bP < 0.01.
Figure 2 SJD inhibited adipogenic differentiation of NSAA-derived BMSCs A: oil Red O staining of lipid droplets (× 40, Scale bar = 50 μm). A1: Control group; A2: 0.08 μg/mL group; A3: 0.8 μg/mL group; A4: 8 μg/mL group; A5: 80 μg/mL group. Control group: basal medium; 0.08 μg/mL group: BMSCs were treated with 0.08 μg/mL SJD for 48 h; 0.8 μg/mL group: BMSCs were treated with 0.8 μg/mL SJD for 48 h; 8 μg/mL group: BMSCs were treated with 8 μg/mL SJD for 48 h; 80 μg/mL group: BMSCs were treated with 80 μg/mL SJD for 48 h; B: relative mRNA levels of PPARG and FABP4 were detected by qRT-PCR. B1: quantitative analysis of PPARG levels; B2: quantitative analysis of FABP4 levels; C: the protein levels of PPARG and FABP4 were detected by Western blot. C1: protein images of PPARG and FABP4; C2: quantitative analysis of PPARG levels; C3: quantitative analysis of FABP4 levels. SJD: Shisiwei Jianzhong decoction; NSAA: non-severe aplastic anemia; BMSCs: bone marrow mesenchymal stem cells; PPARG: peroxisome proliferator-activated receptor gamma; FABP4: fatty acid-binding protein 4; qRT-PCR: quantitative real-time polymerase chain reaction. Statistical analysis was performed using one-way analysis of variance. Data are exhibited as mean ± standard deviation (n = 3). Compared with Control group, aP < 0.05 and bP < 0.01.
Figure 3 SJD downregulated NFATC4 in NSAA-derived BMSCs A: mRNA and protein expression of NFATC4 in BMSCs from NSAA patients and healthy controls. A1: quantitative analysis of NFATC4 mRNA levels; A2: protein images of NFATC4; A3: quantitative analysis of NFATC4 protein levels. Healthy control group: BMSCs isolated from healthy controls; NSAA group: BMSCs isolated from patients with NSAA; n = 6; statistical analysis was performed using t-test; B: mRNA and protein expression of NFATC4 in SJD-treated BMSCs from NSAA patients. B1: quantitative analysis of NFATC4 mRNA levels; B2: protein images of NFATC4; B3: quantitative analysis of NFATC4 protein levels. Control group: basal medium; 0.08 μg/mL group: BMSCs were treated with 0.08 μg/mL SJD for 48 h; 0.8 μg/mL group: BMSCs were treated with 0.8 μg/mL SJD for 48 h; 8 μg/mL group: BMSCs were treated with 8 μg/mL SJD for 48 h; 80 μg/mL group: BMSCs were treated with 80 μg/mL SJD for 48 h. SJD: Shisiwei Jianzhong decoction; NFATC4: nuclear factor of activated T cells, cytoplasmic 4; NSAA: non-severe aplastic anemia; BMSCs: bone marrow mesenchymal stem cells. Statistical analysis was performed using one-way analysis of variance. Data are exhibited as mean ± standard deviation (n = 3). Compared with Healthy control (A) or Control (B), aP < 0.01 and bP < 0.05.
Figure 3 SJD downregulated NFATC4 in NSAA-derived BMSCs A: mRNA and protein expression of NFATC4 in BMSCs from NSAA patients and healthy controls. A1: quantitative analysis of NFATC4 mRNA levels; A2: protein images of NFATC4; A3: quantitative analysis of NFATC4 protein levels. Healthy control group: BMSCs isolated from healthy controls; NSAA group: BMSCs isolated from patients with NSAA; n = 6; statistical analysis was performed using t-test; B: mRNA and protein expression of NFATC4 in SJD-treated BMSCs from NSAA patients. B1: quantitative analysis of NFATC4 mRNA levels; B2: protein images of NFATC4; B3: quantitative analysis of NFATC4 protein levels. Control group: basal medium; 0.08 μg/mL group: BMSCs were treated with 0.08 μg/mL SJD for 48 h; 0.8 μg/mL group: BMSCs were treated with 0.8 μg/mL SJD for 48 h; 8 μg/mL group: BMSCs were treated with 8 μg/mL SJD for 48 h; 80 μg/mL group: BMSCs were treated with 80 μg/mL SJD for 48 h. SJD: Shisiwei Jianzhong decoction; NFATC4: nuclear factor of activated T cells, cytoplasmic 4; NSAA: non-severe aplastic anemia; BMSCs: bone marrow mesenchymal stem cells. Statistical analysis was performed using one-way analysis of variance. Data are exhibited as mean ± standard deviation (n = 3). Compared with Healthy control (A) or Control (B), aP < 0.01 and bP < 0.05.
Figure 4 NFATC4 overexpression reversed the effects of SJD on adipogenic differentiation and PGC-1α acetylation in NSAA-derived BMSCs A: relative mRNA expression of NFATC4 in SJD-treated BMSCs was detected by qRT-PCR. Control group: basal medium; CsA group: CsA (1 μM); SJD group: SJD (80 μg/mL); SJD + oe-NC group: SJD (80 μg/mL) treated with oe-NC BMSCs; SJD + oe-NFATC4 group: SJD (80 μg/mL) treated with oe-NFATC4 BMSCs; B: protein levels of NFATC4, PGC-1α, and Ac-PGC-1α in SJD-treated BMSCs were detected by Western blot. B1: protein images of NFATC4, PGC-1α, and Ac-PGC-1α; B2: quantitative analysis of NFATC4 protein levels; B3: quantitative analysis of PGC-1α protein levels; B4: quantitative analysis of Ac-PGC-1α protein levels. C: oil red O staining of lipid droplets (× 40, Scale bar = 50 μm). C1: Control group; C2: CsA group; C3: SJD group; C4: SJD + oe-NC group; C5: SJD + oe-NFATC4 group. NFATC4: nuclear factor of activated T cells, cytoplasmic 4; SJD: Shisiwei Jianzhong decoction; PGC-1α: peroxisome proliferator-activated receptor-gamma coactivator; NSAA: non-severe aplastic anemia; BMSCs: bone marrow mesenchymal stem cells; qRT-PCR: quantitative real-time polymerase chain reaction; CsA: cyclosporine A. Statistical analysis was performed using one-way analysis of variance. Data are exhibited as mean ± standard deviation (n = 3). Compared with control group, aP < 0.01 and cP < 0.05; compared with SJD + oe-NC group, bP < 0.01.
Figure 4 NFATC4 overexpression reversed the effects of SJD on adipogenic differentiation and PGC-1α acetylation in NSAA-derived BMSCs A: relative mRNA expression of NFATC4 in SJD-treated BMSCs was detected by qRT-PCR. Control group: basal medium; CsA group: CsA (1 μM); SJD group: SJD (80 μg/mL); SJD + oe-NC group: SJD (80 μg/mL) treated with oe-NC BMSCs; SJD + oe-NFATC4 group: SJD (80 μg/mL) treated with oe-NFATC4 BMSCs; B: protein levels of NFATC4, PGC-1α, and Ac-PGC-1α in SJD-treated BMSCs were detected by Western blot. B1: protein images of NFATC4, PGC-1α, and Ac-PGC-1α; B2: quantitative analysis of NFATC4 protein levels; B3: quantitative analysis of PGC-1α protein levels; B4: quantitative analysis of Ac-PGC-1α protein levels. C: oil red O staining of lipid droplets (× 40, Scale bar = 50 μm). C1: Control group; C2: CsA group; C3: SJD group; C4: SJD + oe-NC group; C5: SJD + oe-NFATC4 group. NFATC4: nuclear factor of activated T cells, cytoplasmic 4; SJD: Shisiwei Jianzhong decoction; PGC-1α: peroxisome proliferator-activated receptor-gamma coactivator; NSAA: non-severe aplastic anemia; BMSCs: bone marrow mesenchymal stem cells; qRT-PCR: quantitative real-time polymerase chain reaction; CsA: cyclosporine A. Statistical analysis was performed using one-way analysis of variance. Data are exhibited as mean ± standard deviation (n = 3). Compared with control group, aP < 0.01 and cP < 0.05; compared with SJD + oe-NC group, bP < 0.01.
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