Journal of Southern Medical University ›› 2025, Vol. 45 ›› Issue (5): 986-994.doi: 10.12122/j.issn.1673-4254.2025.05.11
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Xiaotao LIANG1(), Yifan XIONG1, Xueqi LIU5, Xiaoshan LIANG4, Xiaoyu ZHU1, Wei XIE1,2,3,4(
)
Received:
2025-01-23
Online:
2025-05-20
Published:
2025-05-23
Contact:
Wei XIE
E-mail:1092296938@qq.com;xieweizn@126.com
About author:
First author contact:LIANG Xiaotao1, XIONG Yifan1, LIU Xueqi5, LIANG Xiaoshan4, ZHU Xiaoyu1, XIE Wei1, 2, 3, 4
Xiaotao LIANG, Yifan XIONG, Xueqi LIU, Xiaoshan LIANG, Xiaoyu ZHU, Wei XIE. Huoxue Shufeng Granule alleviates central sensitization in chronic migraine mice via TLR4/NF-κB inflammatory pathway[J]. Journal of Southern Medical University, 2025, 45(5): 986-994.
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URL: https://www.j-smu.com/EN/10.12122/j.issn.1673-4254.2025.05.11
Primer name | Sequence |
---|---|
IFN-γ Forward | AGCAACAACATAAGCGTCATTGA |
IFN-γ Reverse | TGACCTCAAACTTGGCAATACTC |
TNF-α Forward | GCCTCCCTCTCATCAGTTCTATG |
TNF-α Reverse | ACCTGGGAGTAGACAAGGTACAA |
IL-1β Forward | GCAACTGTTCCTGAACTCAACT |
IL-1β Reverse | ATCTTTTGGGGTCCGTCAACT |
IL-6 Forward | ACAACCACGGCCTTCCCTACTT |
IL-6 Reverse | CACGATTTCCCAGAGAACATGTG |
GAPDH Forward | CAACTACATGGTCTACATGTTC |
GAPDH Reverse | CTCGCTCCTGGAAGATG |
Tab.1 Primer sequences used in this study
Primer name | Sequence |
---|---|
IFN-γ Forward | AGCAACAACATAAGCGTCATTGA |
IFN-γ Reverse | TGACCTCAAACTTGGCAATACTC |
TNF-α Forward | GCCTCCCTCTCATCAGTTCTATG |
TNF-α Reverse | ACCTGGGAGTAGACAAGGTACAA |
IL-1β Forward | GCAACTGTTCCTGAACTCAACT |
IL-1β Reverse | ATCTTTTGGGGTCCGTCAACT |
IL-6 Forward | ACAACCACGGCCTTCCCTACTT |
IL-6 Reverse | CACGATTTCCCAGAGAACATGTG |
GAPDH Forward | CAACTACATGGTCTACATGTTC |
GAPDH Reverse | CTCGCTCCTGGAAGATG |
Fig.2 HXSFG alleviates baseline and acute mechanical and thermal hyperalgesia in mice with NTG-induced CM. A: Changes in baseline mechanical pain thresholds of the hind paws of the mice. B: Changes in acute mechanical pain thresholds of the hind paws of the mice. C: Changes in baseline thermal pain thresholds of the hind paws. D: Changes in acute thermal pain thresholds of the hind paws. Data are presented as Mean±SE (n=12). Statistical analysis was performed using two-way ANOVA followed by Tukey's post hoc test. **P<0.01 Model group vs Control group; #P<0.05, ##P<0.01 Topiramate group vs Model group; &P<0.05, &&P<0.01 HXSFG-L group vs Model group; ^^P<0.01 HXSFG-H group vs Model group.
Fig.3 Effects of HXSFG on c-Fos expression levels in the TNC region of mice with NTG-induced CM. A: Immunofluorescence images of c-Fos expression (Scale bar: 100 μm). B: Quantitative analysis of c-Fos immunoreactivity (Each group included 4 mice, and 3 slices were analyzed for each mouse). C: Protein immunoblot images of c-Fos expression. D: Quantitative analysis of c-Fos protein immunoblotting (n=4). Data are presented as Mean±SE. Statistical analysis was performed using two-way ANOVA followed by Tukey's post hoc test. **P<0.01 vs Control group; #P<0.05, ##P<0.01 vs Model group; &P<0.05, &&P<0.01 vs Model group; ^^P<0.01 vs Model group.
Fig.4 Effects of HXSFG on CGRP expression levels in the TNC region of mice with NTG-induced CM. A: Immunofluorescence images of CGRP expression (×40, scale bar: 500 μm). B: Quantitative analysis of CGRP immunoreactivity (Each group included 4 mice, and 3 slices were analyzed for each mouse). C: Protein immunoblot images of CGRP expression. D: Quantitative analysis of CGRP protein immunoblotting (n=4). Data are presented as Mean±SE. Statistical analysis was performed using two-way ANOVA followed by Tukey's post hoc test. **P<0.01 vs Control group; ##P<0.01 vs Model group; &&P<0.01 vs Model group; ^^P<0.01 vs Model group.
Fig.5 Effects of HXSFG on mRNA levels of inflammatory factors in the TNC region of mice with NTG-induced CM. A: Relative mRNA level of IL-1β. B: Relative mRNA level of IL-6. C: Relative mRNA level of TNF‑α. D: Relative mRNA level of IFN-γ. **P<0.01 vs Control group; ##P<0.01 vs Model group; &&P<0.01 vs Model group; ^^P<0.01 vs Model group.
Fig. 6 Effects of HXSFG on the TLR4/NF-κB signaling pathway in the TNC region of mice with NTG-induced CM. A: Protein immunoblot images of TLR4, p-NF-κB, NF-κB, IL-1β, TNF-α expression. B: Quantitative analysis of TLR4 protein immunoblotting. C: Quantitative analysis of p-NF-κB/NF-κB protein immunoblotting. D: Quantitative analysis of IL-1β protein immunoblotting. E: Quantitative analysis of TNF-α protein immunoblotting (n=4). Data are presented as Mean±SE. Statistical analysis was performed using two-way ANOVA followed by Tukey's post hoc test. **P<0.01 vs Control group; ##P<0.01 vs Model group; &&P<0.01 vs Model group; ^^P<0.01 vs Model group.
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