1.Investigation of the Mechanism of Cold Hyperalgesia in KOA Mice Relieved by Shangke Lengtongtie Based on HMGB1/CX-CL12/CXCR4 Signaling Axis
Yibao WEI ; Li ZHANG ; Taiyang LIAO ; Lishi JIE ; Zhenyuan MA ; Peng WU ; Zhengquan HUANG ; Li ZHANG ; Liang DING ; Wei MEI ; Runlin XING ; Songjiang YIN ; Xiaochen LI ; Nongshan ZHANG ; Jun MAO ; Pei-min WANG
Journal of Nanjing University of Traditional Chinese Medicine 2025;41(2):195-202
OBJECTIVE To explore the intervention mechanism of Shangke Lengtongtie on cold hyperalgesia in KOA mice based on the HMGB1/CXCL12/CXCR4 signaling axis.METHODS Monosodium iodoacetate(MIA)was used for the intra-articular injec-tion into the knee joint to establish mice model of knee osteoarthritis(KOA).Peripheral blood monocytes were extracted from mice,cultured,and then reinfused into the tail vein of the mice.Subsequently,in vivo animal imaging was used to observe the recruitment sites of these monocytes.The cold hyperalgesia threshold was measured at various time points in each group of mice.Hematoxylin and eosin(HE)staining was used to evaluate the level of synovial pathological changes.ELISA was employed to detect the expression of in-flammatory factors IL-1β,TNF-α,and pain mediators CGRP and Substance P in mouse serum.Western blot and qPCR methods were used to detect the protein and gene expression of cold hyperalgesia-related indicators such as TRPA1,TRPM8,HMGB1,CXCL12,CXCR4,Collagen Ⅰ,and Netrin-1 in synovial tissue,as well as DCC in dorsal root ganglia(DRG)tissue.RESULTS In vivo ima-ging showed that after the monocytes were reinfused into KOA mice,they were recruited to the knee joint area,with the HMGB1 group exhibiting a greater recruitment of circulating monocytes at the knee joint.Additionally,compared to the control group,the KOA group and HMGB1 group showed inflammatory pathological changes in the synovium,increased expression of serum inflammatory factors and pain mediators,reduced cold hyperalgesia threshold,and upregulated protein and gene expression of cold hyperalgesia-related indica-tors in synovial and DRG tissues.The changes were more significant in the HMGB1 group compared to the KOA group(P<0.05).Af-ter treatment with Shangke Lengtongtie or GL intervention,synovial inflammation was alleviated,serum inflammatory factors and pain mediators decreased,cold hyperalgesia threshold increased,and the upregulation of cold hyperalgesia-related indicator protein and gene expression levels was significantly reversed(P<0.05).CONCLUSION Shangke Lengtongtie exerts a beneficial effect on the mitigation of synovitis and cold hyperalgesia in KOA mice,a therapeutic mechanism that possibly mediated through the inhibition of the HMGB1/CXCL12/CXCR4 signaling axis.
2.Investigation of the Mechanism of Cold Hyperalgesia in KOA Mice Relieved by Shangke Lengtongtie Based on HMGB1/CX-CL12/CXCR4 Signaling Axis
Yibao WEI ; Li ZHANG ; Taiyang LIAO ; Lishi JIE ; Zhenyuan MA ; Peng WU ; Zhengquan HUANG ; Li ZHANG ; Liang DING ; Wei MEI ; Runlin XING ; Songjiang YIN ; Xiaochen LI ; Nongshan ZHANG ; Jun MAO ; Pei-min WANG
Journal of Nanjing University of Traditional Chinese Medicine 2025;41(2):195-202
OBJECTIVE To explore the intervention mechanism of Shangke Lengtongtie on cold hyperalgesia in KOA mice based on the HMGB1/CXCL12/CXCR4 signaling axis.METHODS Monosodium iodoacetate(MIA)was used for the intra-articular injec-tion into the knee joint to establish mice model of knee osteoarthritis(KOA).Peripheral blood monocytes were extracted from mice,cultured,and then reinfused into the tail vein of the mice.Subsequently,in vivo animal imaging was used to observe the recruitment sites of these monocytes.The cold hyperalgesia threshold was measured at various time points in each group of mice.Hematoxylin and eosin(HE)staining was used to evaluate the level of synovial pathological changes.ELISA was employed to detect the expression of in-flammatory factors IL-1β,TNF-α,and pain mediators CGRP and Substance P in mouse serum.Western blot and qPCR methods were used to detect the protein and gene expression of cold hyperalgesia-related indicators such as TRPA1,TRPM8,HMGB1,CXCL12,CXCR4,Collagen Ⅰ,and Netrin-1 in synovial tissue,as well as DCC in dorsal root ganglia(DRG)tissue.RESULTS In vivo ima-ging showed that after the monocytes were reinfused into KOA mice,they were recruited to the knee joint area,with the HMGB1 group exhibiting a greater recruitment of circulating monocytes at the knee joint.Additionally,compared to the control group,the KOA group and HMGB1 group showed inflammatory pathological changes in the synovium,increased expression of serum inflammatory factors and pain mediators,reduced cold hyperalgesia threshold,and upregulated protein and gene expression of cold hyperalgesia-related indica-tors in synovial and DRG tissues.The changes were more significant in the HMGB1 group compared to the KOA group(P<0.05).Af-ter treatment with Shangke Lengtongtie or GL intervention,synovial inflammation was alleviated,serum inflammatory factors and pain mediators decreased,cold hyperalgesia threshold increased,and the upregulation of cold hyperalgesia-related indicator protein and gene expression levels was significantly reversed(P<0.05).CONCLUSION Shangke Lengtongtie exerts a beneficial effect on the mitigation of synovitis and cold hyperalgesia in KOA mice,a therapeutic mechanism that possibly mediated through the inhibition of the HMGB1/CXCL12/CXCR4 signaling axis.
3.Determination of Feruloyltyramine in Pothi Chinensis Herba by HPLC
Wenhui LUO ; Buming LIU ; Xiaoli BI ; Lishi PENG ; Hongcong QIU
China Pharmacist 2015;18(10):1790-1792
Objective: To develop an HPLC method for determining feruloyltyramine in Pothi chinensis herba. Methods: The HPLC determination was performed on a Thermo ODS-2 Hypersil column(250 mm × 4. 6 mm, 5 μm) with isocratic elution of metha-nol-0. 4% phosphoric (35:65) as the mobile phase. The flow rate was 1. 0 ml·min-1 . The column temperature was at 25℃, and the detection wavelength was set at 318 nm. Results: Feruloyltyramine showed good linearity,and the recovery was 99. 40% with RSD of 1. 44% (n =9). Conclusion: The method is quick, simple and reproducible, which can be used to control the quality of Pothi chinensis herba.

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