Journal of Traditional Chinese Medicine ›› 2024, Vol. 44 ›› Issue (3): 437-447.DOI: 10.19852/j.cnki.jtcm.20240423.004
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Transient receptor potential vanilloid 1 involved in the analgesic effects of total flavonoids extracted from Longxuejie (Resina Dracaenae Cochinchinensis)
MO Xiaoqiang1,2, CHEN Yating3, YIN Qian3, CHEN Haibo1, BAN Qiang1, LI Jun4, CHEN Su3(
), YAO Jinguang1()
- 1 Department of Stomatology, Affiliated Hospital of Youjiang Medical University for Nationalities, Baise 533000, China
2 College of General Practitioners, Youjiang Medical University for Nationalities, Baise 533000, China
3 College of Biomedical Engineering, South-Central Minzu University, Key Laboratory of Cognitive Science of State Ethnic Affairs Commission, Hubei Key Laboratory of Medical Information Analysis and Tumor Diagnosis and Treatment, Wuhan 430074, China
4 College of Pharmacy, South-Central Minzu University, Wuhan 430074, China
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Received:2023-02-22Accepted:2023-07-06Online:2024-06-15Published:2024-04-30 -
Contact:CHEN Su,YAO Jinguang -
About author:Prof. CHEN Su, College of Biomedical Engineering, South-Central Minzu University, Wuhan 430074, China. ecloud.7@163.com Telephone: +86-27-67841502
Prof. YAO Jinguang, Affiliated Hospital of Youjiang Medical University for Nationalities, Baise 533000, China. yao7760698@126.com;
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Supported by:High Level Talents Project of Affiliated Hospital of Youjiang Medical University for Nationalities: Study of Soft-Du'an Capsule's Mechanism and Efficacy of Regulating TRPV1 Pashways in Relieving Oral and Maxillofacial Trigeminal Neuralgia(YYFYR20213002);Innovative Group Project of Natural Science Foundation of Hubei Province: Study on the Mechanisms of Pain Signal Transduction and Drug Analgesia(2020CFA025)
Cite this article
MO Xiaoqiang, CHEN Yating, YIN Qian, CHEN Haibo, BAN Qiang, LI Jun, CHEN Su, YAO Jinguang. Transient receptor potential vanilloid 1 involved in the analgesic effects of total flavonoids extracted from Longxuejie (Resina Dracaenae Cochinchinensis)[J]. Journal of Traditional Chinese Medicine, 2024, 44(3): 437-447.
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Figure 1 Chemical structures of 1-12 and the typical HPLC chromatographic profile of TFDB A: chemical structures of 1-12 from TFDB. Twelve compounds are Kumatakemin B (1), Resveratrol (2), Loureirin D (3), 7,4'-Dihydroxydihydrohomoisoflavone (4), Loureirin C (5), 4,4'-Dihydroxy-2,6-dimethoxydihydrochalcone (6), 7,4'-Dihydroxyflavan (7), 3,4'-dihydroxy-5-methoxystilbene (8), Cochinchinenin B (9), Loureirin A (10), Loureirin B (11) and Pterostilbene (12); B: typical HPLC chromatographic profile of TFDB. The typical HPLC chromatographic profile of TFDB. HPLC peaks: 1 (11.633 min); 2 (12.093 min); 3 (14.460 min); 4 (17.073 min); 5 (19.760 min); 6 (21.413 min); 7 (24.827 min); 8 (27.147 min); 9 (28.387 min); 10 (50.587 min); 11 (52.080 min); 12 (62.320 min). HPLC: high performance liquid chromatography; TFDB: total flavonoids of dragon's blood.
Figure 1 Chemical structures of 1-12 and the typical HPLC chromatographic profile of TFDB A: chemical structures of 1-12 from TFDB. Twelve compounds are Kumatakemin B (1), Resveratrol (2), Loureirin D (3), 7,4'-Dihydroxydihydrohomoisoflavone (4), Loureirin C (5), 4,4'-Dihydroxy-2,6-dimethoxydihydrochalcone (6), 7,4'-Dihydroxyflavan (7), 3,4'-dihydroxy-5-methoxystilbene (8), Cochinchinenin B (9), Loureirin A (10), Loureirin B (11) and Pterostilbene (12); B: typical HPLC chromatographic profile of TFDB. The typical HPLC chromatographic profile of TFDB. HPLC peaks: 1 (11.633 min); 2 (12.093 min); 3 (14.460 min); 4 (17.073 min); 5 (19.760 min); 6 (21.413 min); 7 (24.827 min); 8 (27.147 min); 9 (28.387 min); 10 (50.587 min); 11 (52.080 min); 12 (62.320 min). HPLC: high performance liquid chromatography; TFDB: total flavonoids of dragon's blood.
Figure 2 Effects of TFDB on capsaicin-induced TRPV1 currents in DRG neurons A: effects of capsazepine on capsaicin-induced TRPV1 currents in DRG neurons. Capsazepine (10 μmol/L) inhibited rapidly the current induced by 1 μmol/L capsaicin, consistent with activation of TRPV1 receptor. B: concentration-response curves for the inhibition on the capsaicin-induced TRPV1 currents of TFDB. Data points represented the percent change in peak amplitude in the presence of TFDB (each concentration n = 7, one-way analysis of variance with Dunnett-t test, P < 0.05). The data points are fitted with Hill's Equation. C: effect of TFDB on capsaicin-induced TRPV1 currents in DRG neurons. TFDB (1, 3, 10, 30, 100 μg/mL) inhibited rapidly and reversibly the current induced by 1 μmol/L capsaicin. CAP: capsaicin; CPZ: capsazepine; TFDB: total flavonoids of dragon's blood; TRPV1: transient receptor potential vanilloid 1; DRG: dorsal root ganglion.
Figure 2 Effects of TFDB on capsaicin-induced TRPV1 currents in DRG neurons A: effects of capsazepine on capsaicin-induced TRPV1 currents in DRG neurons. Capsazepine (10 μmol/L) inhibited rapidly the current induced by 1 μmol/L capsaicin, consistent with activation of TRPV1 receptor. B: concentration-response curves for the inhibition on the capsaicin-induced TRPV1 currents of TFDB. Data points represented the percent change in peak amplitude in the presence of TFDB (each concentration n = 7, one-way analysis of variance with Dunnett-t test, P < 0.05). The data points are fitted with Hill's Equation. C: effect of TFDB on capsaicin-induced TRPV1 currents in DRG neurons. TFDB (1, 3, 10, 30, 100 μg/mL) inhibited rapidly and reversibly the current induced by 1 μmol/L capsaicin. CAP: capsaicin; CPZ: capsazepine; TFDB: total flavonoids of dragon's blood; TRPV1: transient receptor potential vanilloid 1; DRG: dorsal root ganglion.
Figure 3 Effects of TFDB in different pain models (formalin test, capsaicin test and CFA-induced inflammation pain rats) A: effect of TFDB against formalin-induced licking in the first phase in mice; B: effect of TFDB against formalin-induced licking in the second phase in mice; C: effect of TFDB against capsaicin-induced licking in mice. D: short term analgesic effect of TFDB on CFA-induced inflammatory pain rats. Paw-withdrawal latencies before and after 48 h CFA administrated. After CFA-induced inflammatory pain model rats were successfully established, paw-withdrawal latencies after 30, 60 and 90 min drug administrated; E: Long term analgesic effect of TFDB on CFA-induced inflammatory pain rats. Paw-withdrawal latencies before and after 48 h CFA administrated. After CFA-induced inflammatory pain model rats were successfully established, paw-withdrawal latencies after 1, 2 and 3 d drug administrated. TFDB: total flavonoids of dragon's blood; CFA: complete freunds adjuvant; SEM: standard error of the mean; ANOVA: analysis of variance. Each column or point represented the mean of the values obtained in 10 mice and the error bars indicate the SEM. Statistical analysis was performed using one-way ANOVA with Dunnett-t test. aP < 0.01 vs control, bP < 0.05 vs control.
Figure 3 Effects of TFDB in different pain models (formalin test, capsaicin test and CFA-induced inflammation pain rats) A: effect of TFDB against formalin-induced licking in the first phase in mice; B: effect of TFDB against formalin-induced licking in the second phase in mice; C: effect of TFDB against capsaicin-induced licking in mice. D: short term analgesic effect of TFDB on CFA-induced inflammatory pain rats. Paw-withdrawal latencies before and after 48 h CFA administrated. After CFA-induced inflammatory pain model rats were successfully established, paw-withdrawal latencies after 30, 60 and 90 min drug administrated; E: Long term analgesic effect of TFDB on CFA-induced inflammatory pain rats. Paw-withdrawal latencies before and after 48 h CFA administrated. After CFA-induced inflammatory pain model rats were successfully established, paw-withdrawal latencies after 1, 2 and 3 d drug administrated. TFDB: total flavonoids of dragon's blood; CFA: complete freunds adjuvant; SEM: standard error of the mean; ANOVA: analysis of variance. Each column or point represented the mean of the values obtained in 10 mice and the error bars indicate the SEM. Statistical analysis was performed using one-way ANOVA with Dunnett-t test. aP < 0.01 vs control, bP < 0.05 vs control.
Figure 4 Effects of TFDB on TRPV1 expression in DRG neurons of CFA-induced inflammation pain rats A: an example of Western blot images for TRPV1 and GAPDH of all drug administration groups; B: standardization ratio of TRPV1 to GAPDH band densities in DRG neurons of various drugs-treated CFA-induced inflammation pain rats (Six batches for all the groups). C: fluorescence optical intensity of TRPV1 protein (Five random visual field images at × 400 magnification for each sample); D: Immunofluorescence showed the expression of TRPV1 receptor protein in DRG neurons. Scale bar = 50 μm; D1: DRG neurons immunofluorescence image of normal saline group; D2: DRG neurons immunofluorescence image of aspirin group; D3: DRG neurons immunofluorescence image of TFDB group. DRG neurons were derived from three groups of rats. Three groups of 10 rats were respectively treated with the same volume of normal saline (10 mL/kg), aspirin (20 mg/kg) and TFDB (20 mg/kg). TFDB: total flavonoids of dragon's blood; TRPV1: transient receptor potential vanilloid 1; DRG: dorsal root ganglion; CFA: complete freunds adjuvant; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; ANOVA: analysis of variance; SEM: standard error of the mean. Data were expressed as the mean ± SEM. Statistical analysis was performed using one-way ANOVA with Dunnett-t test. aP < 0.001 vs control.
Figure 4 Effects of TFDB on TRPV1 expression in DRG neurons of CFA-induced inflammation pain rats A: an example of Western blot images for TRPV1 and GAPDH of all drug administration groups; B: standardization ratio of TRPV1 to GAPDH band densities in DRG neurons of various drugs-treated CFA-induced inflammation pain rats (Six batches for all the groups). C: fluorescence optical intensity of TRPV1 protein (Five random visual field images at × 400 magnification for each sample); D: Immunofluorescence showed the expression of TRPV1 receptor protein in DRG neurons. Scale bar = 50 μm; D1: DRG neurons immunofluorescence image of normal saline group; D2: DRG neurons immunofluorescence image of aspirin group; D3: DRG neurons immunofluorescence image of TFDB group. DRG neurons were derived from three groups of rats. Three groups of 10 rats were respectively treated with the same volume of normal saline (10 mL/kg), aspirin (20 mg/kg) and TFDB (20 mg/kg). TFDB: total flavonoids of dragon's blood; TRPV1: transient receptor potential vanilloid 1; DRG: dorsal root ganglion; CFA: complete freunds adjuvant; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; ANOVA: analysis of variance; SEM: standard error of the mean. Data were expressed as the mean ± SEM. Statistical analysis was performed using one-way ANOVA with Dunnett-t test. aP < 0.001 vs control.
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