Journal of Traditional Chinese Medicine ›› 2024, Vol. 44 ›› Issue (3): 489-495.DOI: 10.19852/j.cnki.jtcm.20231204.005
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FENG Yanchen1, LIU Yali1, DANG Xue1, LIN Zixuan2, ZHANG Yunke2, CHE Zhiying1, LI Xiang3, PAN Xiaolong1, LIU Feixiang2(), ZHENG Pan1()
Received:
2023-02-22
Accepted:
2023-07-14
Online:
2024-06-15
Published:
2023-12-04
Contact:
LIU Feixiang, Hospital of Encephalopathy, the First Affiliated Hospital of Henan University of Chinese Medicine, Zhengzhou 450000, China, Supported by:
FENG Yanchen, LIU Yali, DANG Xue, LIN Zixuan, ZHANG Yunke, CHE Zhiying, LI Xiang, PAN Xiaolong, LIU Feixiang, ZHENG Pan. Exploring the multicomponent synergy mechanism of Zuogui Wan (左归丸) on postmenopausal osteoporosis by a systems pharmacology strategy[J]. Journal of Traditional Chinese Medicine, 2024, 44(3): 489-495.
Figure 1 Effects of ZGW on bone-related indexes in rats A: serum levels of BALP were evaluated using ELISA kits (n = 5); B: serum levels of BALP and β-CTX were evaluated using ELISA kits (n = 5); C: changes in BMD levels in rats from each group following ZGW intervention (n = 7); D: changes in the levels of the area under the curve of the femoral stem (n = 4); E: changes in the levels of the maximum load of the femoral stem (n = 4); F: changes in the levels of the stiffness coefficient of the femoral stem (n = 4). Sham: sham-operated, treated with water for 12 weeks; OVX: ovariectomized, treated with water for 12 weeks; E2: 17β-estradiol, treated with 17β-E2 for 12 weeks (50 μg·kg-1·d-1); ZGW-L: low-dose ZGW (2.3 g·kg-1·d-1), treated with low-dose lyophilized ZGW powder dissolved in water for 12 weeks; ZGW-H: high-dose ZGW (4.6 g·kg-1·d-1), treated with high-dose lyophilized ZGW powder dissolved in water for 12 weeks. ZGW: Zuogui Wan. BALP: bone-specific alkaline phosphatase. β-CTX: β-C-terminal telopeptide of type I collagen. ELISA: enzyme-linked immunosorbent assay. BMD: bone mineral density. dP < 0.05, compared with the Sham group; aP < 0.01, compared with the Sham group; cP < 0.05, compared with the OVX group; bP < 0.01, compared with the OVX group. Data were presented as mean ± standard error of the mean. One-way Analysis of variance, followed by Student-Newman-Keuls test and the Student's t-test.
Figure 2 Changes in HE staining of the distal femur in groups after ZGW intervention (× 40) A: Sham group (n = 4). B: OVX group (n = 4). C: E2 group (n = 4). D: ZGW-L group (n = 4). E: ZGW-G group (n = 4). Sham: sham-operated, treated with water for 12 weeks; OVX: ovariectomized, treated with water for 12 weeks; E2: 17β-estradiol, treated with 17β-E2 for 12 weeks (50 μg·kg-1·d-1); ZGW-L: low-dose ZGW (2.3 g·kg-1·d-1), treated with low-dose lyophilized ZGW powder dissolved in water for 12 weeks; ZGW-H: high-dose ZGW (4.6 g·kg-1·d-1), treated with high-dose lyophilized ZGW powder dissolved in water for 12 weeks. HE: hematoxylin-eosin. ZGW: Zuogui Wan.
Figure 3 Changes in core gene expression levels after ZGW intervention A: protein expression levels of ADRA2A (n = 3); B: protein expression levels of CXCL8 (n = 3); C: protein expression levels of Runx2 (n = 3); D: protein expression levels of CCR2 (n = 3); E: protein expression levels of APP (n = 3); F: protein expression levels of MTNR1B (n = 3); G: Western blotting representative images of ADRA2A, CXCL8, RUNX2, CCR2, APP and MTNR1B (n = 3). Sham: sham-operated, treated with water for 12 weeks; OVX: ovariectomized, treated with water for 12 weeks; E2: 17β-estradiol, treated with 17β-E2 for 12 weeks (50 μg·kg-1·d-1); ZGW-L: low-dose ZGW (2.3 g·kg-1·d-1), treated with low-dose lyophilized ZGW powder dissolved in water for 12 weeks; ZGW-H: high-dose ZGW (4.6 g·kg-1·d-1), treated with high-dose lyophilized ZGW powder dissolved in water for 12 weeks. ZGW: Zuogui Wan; ADRA2A: rabbit anti-alpha-2a active receptor; CXCL8: rabbit anti-chemokine living 8; RUNX2: runt-related transcription factor 2; CCR2: C-C chemokine receptor type 2; APP: amyloid-beta A4 protein; MTNR1B: melatonin receptor type 1B. aP < 0.01, dP < 0.05, compared with Sham group; bP < 0.01, cP < 0.05, compared with OVX group. Data were presented as mean ± standard error of the mean. One-way Analysis of variance, followed by Student-Newman-Keuls test and the Student's t-test.
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