Journal of Traditional Chinese Medicine ›› 2023, Vol. 43 ›› Issue (3): 533-541.DOI: 10.19852/j.cnki.jtcm.2023.03.003
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JIA Lihua1(), KUANG Haodan2, XU Yuan3()
Received:
2021-10-12
Accepted:
2022-01-27
Online:
2023-06-15
Published:
2023-04-28
Contact:
XU Yuan, Ningbo medical center Lihuili Hospital, Ningbo 315100, China. xu565831937@163.com. Telephone: +86-574-87018617JIA Lihua, KUANG Haodan, XU Yuan. Efficacy of Buzhong Yiqi decoction (补中益气汤) on benign prostatic hyperplasia and its possible mechanism[J]. Journal of Traditional Chinese Medicine, 2023, 43(3): 533-541.
Gene | Sequence |
---|---|
IL-6 | F: 5′-TGGCTGCAGGACATGACAACT-3′ R: 5′-ATCTGAGGTGCCCATGCTACA-3′ |
IL-1β | F: 5′-CTGATGGCCCTAAACAGATGAAG-3′ R: 5′-GGTCGGAGATTCGTAGCAGCTGGAT-3′ |
TNF-α | F: 5′-AAACACACGAGACGCTGAAG-3′ R: 5′-ATCCAGTGAGTTCCGAAAGC-3′ |
β-actin | F: 5'-TCTTGGGTATGGAATCCTGTGGCA-3' R: 5'-ACAGCACTGTGTTGGCATAGAGGT-3' |
Table 1 PCR primers
Gene | Sequence |
---|---|
IL-6 | F: 5′-TGGCTGCAGGACATGACAACT-3′ R: 5′-ATCTGAGGTGCCCATGCTACA-3′ |
IL-1β | F: 5′-CTGATGGCCCTAAACAGATGAAG-3′ R: 5′-GGTCGGAGATTCGTAGCAGCTGGAT-3′ |
TNF-α | F: 5′-AAACACACGAGACGCTGAAG-3′ R: 5′-ATCCAGTGAGTTCCGAAAGC-3′ |
β-actin | F: 5'-TCTTGGGTATGGAATCCTGTGGCA-3' R: 5'-ACAGCACTGTGTTGGCATAGAGGT-3' |
Figure 1 BZYQD “Herb-Compound-Target-Disease” network Round yellow nodes stand for putative targets of ingredients contained in BZYQD; Green diamonds stand for herbs; Blue triangle stand for therapeutic targets for BPH; Round red node stands for the disease of BPH. BZYQD: Buzhong yiqi decoction; BPH: benign prostatic hyperplasia.
Figure 2 Target protein interaction network Round nodes stand for therapeutic targets for BPH and the number of edges from Target protein network. BPH: benign prostatic hyperplasia.
Figure 3 Gene function and KEGG pathway enrichment. A: biological process (BP); B: cell component (CC); C: molecular function (MF); D: KEGG pathway enrichment. The X-axis is the magnitude of P value, and the Y-axis is the path name. The size of the bubble chart represented the count number, and the darker the color, the smaller P value.
Figure 4 Molecular docking results of Quercetin and MAPK8 and IL-6 A: the molecular docking result of Quercetin and MAPK8 (P38) (3D). B: the molecular docking result of Quercetin and IL-6 (3D). C: the molecular docking result of Quercetin and MAPK8 (P38) (2D). D: The molecular docking result of Quercetin and IL-6 (2D).
Figure 5 Anti-proliferative effect of quercetin on BPH-1 cells by MTT assay and quercetin inhibited the production of IL-1β, TNF-α, IL-6 A: quercetin was at the dose of 15, 30, 60, 120 μM and incubation time was at 12, 24, 48, 72 h; B-D: productions of interleukin -6 (IL-6), tumor necrosis factor-α (TNF-α), interleukin 1β (IL-1β) in culture supernatants were detected by enzyme-linked immunosorbent assay kit. E-G: expressions of IL-1β, TNF-α, IL-6 mRNA were measured by RT-RCR. Control group: quercetin 0 μM, 24 h; Que-15: quercetin 15 μM, 24 h; Que-60: quercetin 60 μM, 24 h; Data are expressed as mean?±?standard deviation of three independent experiments. aP?<?0.01 compared with Normal group; bP?<?0.01 compared with Model group.
Figure 6 Quercetin inhibited the expression of p-P38 in the BPH-1 cells. A: Western bot was used to detect the expression of p-P38, P38, MMP-9 and β-actin. B-C: The statistical graph of MMP-9 and P38/p-P38 protein expression which were fold over control group. Control group: quercetin 0 μM, 24 h; Que-15: quercetin 15 μM, 24 h; Que-60: quercetin 60 μM, 24 h; Data are expressed as mean?±?standard deviation of three independent experiments. aP?<?0.01 compared with Normal group; bP?<?0.01 compared with Model group.
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