Journal of Traditional Chinese Medicine ›› 2026, Vol. 46 ›› Issue (2): 427-438.DOI: 10.19852/j.cnki.jtcm.2026.02.015
• Original Articles • Previous Articles Next Articles
Effect of Huiyang Shengji unguent (回阳生肌膏) on the NOD-like receptor family pyrin domain containing protein 3/caspase1/ gasdermin D pathway and lymphatic angiogenesis in patients with diabetic foot ulcer
YU Fangning1, LIN Li1, TANG Xiao1, HE Xiujuan3, ZHANG Bo1, CHEN Yukun2, MA Yizhao2, LIU Zeyu1, YE Jinsheng4, XU Xuying1(
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Department of Vascular Surgery ,Beijing Traditional Chinese Medicine Hospital, Capital Medical University Beijing 100010, China
2Clinical Medical College ,Beijing University of Chinese Medicine Beijing 100020, China
3Research Laboratory for the Theory and Application of Wound Healing ,Beijing Institute of Traditional Chinese Medicine Beijing 100010, China
4Department of General Surgery of Beijing Traditional Chinese Medicine Hospital ,Capital Medical University Beijing 100010, China
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Received:2025-01-11Accepted:2025-06-05Online:2026-04-15Published:2026-04-04 -
Contact:XU Xuying, Department of Vascular Surgery, Beijing Traditional Chinese Medicine Hospital, Capital Medical University, Beijing 100010, China. xxy7341@163.com; Telephone: +86-10-87906642 -
About author:YU Fangning and LIN Li are co-first authors and contributed equally to this work
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Supported by:National Natural Science Foundation of China: Mechanism Study of Huiyang Shengji Ointment in Inducing Acute Inflammation in Chronic Diabetic Foot Ulcers and Promoting Wound Healing Based on the Signal Transducer and Activator of Transcription 3/Forkhead Box Protein M1 Pathway(Grant numbers 82174388)
Cite this article
YU Fangning, LIN Li, TANG Xiao, HE Xiujuan, ZHANG Bo, CHEN Yukun, MA Yizhao, LIU Zeyu, YE Jinsheng, XU Xuying. Effect of Huiyang Shengji unguent (回阳生肌膏) on the NOD-like receptor family pyrin domain containing protein 3/caspase1/ gasdermin D pathway and lymphatic angiogenesis in patients with diabetic foot ulcer[J]. Journal of Traditional Chinese Medicine, 2026, 46(2): 427-438.
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Table 1 Comparison of baseline data in the two patient groups ($\stackrel{-}{x}\pm s$)
| Item | HYSJ group (n = 10) | Control group (n = 10) | P value | |
|---|---|---|---|---|
| Gender (n) | Male | 7 | 8 | 0.6 |
| Female | 3 | 2 | ||
| Age (years) | 67.0±10.5 | 70.4±10.7 | 0.5 | |
| Fasting blood glucose (mmol/L) | 6.2±0.8 | 6.3±1.1 | 0.9 | |
| Glycated hemoglobin (%) | 7.1±1.5 | 7.5±1.9 | 0.6 | |
| TP (g/L) | 65.4±9.3 | 68.7±7.0 | 0.4 | |
| ALB (g/L) | 35.8±7.1 | 35.4±4.2 | 0.9 | |
| ALT (U/L) | 18.5±12.4 | 28.4±41.3 | 0.5 | |
| UA (mmol/L) | 7.4±2.5 | 8.2±3.4 | 0.6 | |
| Scr (umol/L) | 67.1±27.1 | 84.8±30.2 | 0.2 | |
| Hgb (g/L) | 109.7±18.0 | 110.3±22.2 | 0.9 | |
Table 1 Comparison of baseline data in the two patient groups ($\stackrel{-}{x}\pm s$)
| Item | HYSJ group (n = 10) | Control group (n = 10) | P value | |
|---|---|---|---|---|
| Gender (n) | Male | 7 | 8 | 0.6 |
| Female | 3 | 2 | ||
| Age (years) | 67.0±10.5 | 70.4±10.7 | 0.5 | |
| Fasting blood glucose (mmol/L) | 6.2±0.8 | 6.3±1.1 | 0.9 | |
| Glycated hemoglobin (%) | 7.1±1.5 | 7.5±1.9 | 0.6 | |
| TP (g/L) | 65.4±9.3 | 68.7±7.0 | 0.4 | |
| ALB (g/L) | 35.8±7.1 | 35.4±4.2 | 0.9 | |
| ALT (U/L) | 18.5±12.4 | 28.4±41.3 | 0.5 | |
| UA (mmol/L) | 7.4±2.5 | 8.2±3.4 | 0.6 | |
| Scr (umol/L) | 67.1±27.1 | 84.8±30.2 | 0.2 | |
| Hgb (g/L) | 109.7±18.0 | 110.3±22.2 | 0.9 | |
Figure 1 Photographs of the wounds in each group and the changes in wound area 14 d after the intervention A: photographs of typical wounds in the HYSJ group; B: photographs of typical wounds in the control group; C: the wound area before intervention and on the 14th day of intervention; A1, B1: the wound morphology before intervention; A2, B2: the wound morphology on the 7th day of intervention; A3, B3: the wound morphology on the 14th day of intervention; A4, B4: the local wound morphology before intervention; A5, B5: the local wound morphology on the 7th day of intervention; A6, B6: the local wound morphology on the 14th day of intervention. HYSJ group: treated with Huiyang Shengji unguent; control group: treated with bFGF. HYSJ: Huiyang Shengji unguent; bFGF: basic fibroblast growth factor. The independent sample t-test was used for between-group comparisons. Data were expressed as mean ± standard deviation (n = 10). Compared with control group, aP < 0.001.
Figure 1 Photographs of the wounds in each group and the changes in wound area 14 d after the intervention A: photographs of typical wounds in the HYSJ group; B: photographs of typical wounds in the control group; C: the wound area before intervention and on the 14th day of intervention; A1, B1: the wound morphology before intervention; A2, B2: the wound morphology on the 7th day of intervention; A3, B3: the wound morphology on the 14th day of intervention; A4, B4: the local wound morphology before intervention; A5, B5: the local wound morphology on the 7th day of intervention; A6, B6: the local wound morphology on the 14th day of intervention. HYSJ group: treated with Huiyang Shengji unguent; control group: treated with bFGF. HYSJ: Huiyang Shengji unguent; bFGF: basic fibroblast growth factor. The independent sample t-test was used for between-group comparisons. Data were expressed as mean ± standard deviation (n = 10). Compared with control group, aP < 0.001.
Figure 2 HE staining images of the wound tissues from each group at different intervention time A: HE staining of wound tissue in the control group on days 0, 7, and 14 of treatment (× 40); B: HE staining of wound tissue in the HYSJ group on days 0, 7, and 14 of treatment (× 40); A1, B1: image before treatment; A2, B2: image on day 7 of treatment (× 40); A3, B3: image on day 14 of treatment (× 40); A4, B4: detail image before treatment; A5, B5: detail image on day 7 of treatment; A6, B6: detail image on day 14 of treatment. The arrows indicate infiltrated neutrophils. Scale bar: 80 μm. HYSJ group: treated with Huiyang Shengji unguent; control group: treated with bFGF. HE: hematoxylin-eosin; HYSJ: Huiyang Shengji unguent; bFGF: basic fibroblast growth factor.
Figure 2 HE staining images of the wound tissues from each group at different intervention time A: HE staining of wound tissue in the control group on days 0, 7, and 14 of treatment (× 40); B: HE staining of wound tissue in the HYSJ group on days 0, 7, and 14 of treatment (× 40); A1, B1: image before treatment; A2, B2: image on day 7 of treatment (× 40); A3, B3: image on day 14 of treatment (× 40); A4, B4: detail image before treatment; A5, B5: detail image on day 7 of treatment; A6, B6: detail image on day 14 of treatment. The arrows indicate infiltrated neutrophils. Scale bar: 80 μm. HYSJ group: treated with Huiyang Shengji unguent; control group: treated with bFGF. HE: hematoxylin-eosin; HYSJ: Huiyang Shengji unguent; bFGF: basic fibroblast growth factor.
Table 2 Changes of IL-1β and IL-18 contents in wound secretion of two groups of patients [pg/mL, M (Q1, Q3)]
| Item | Control group (n = 10) | HYSJ group (n = 10) | |
|---|---|---|---|
| IL-1β | Day 0 | 2472.35 (379.63, 82825.95) | 2366.69 (1011.29, 15433.128) |
| Day 7 | 1864.33 (994.49, 2583.92) | 604.83 (367.05, 1169.90)a | |
| Day 14 | 2035.07 (875.39, 2933.65) | 548.94 (115.37, 692.52)ab | |
| IL-18 | Day 0 | 1073.13 (412.47, 4554.46) | 2036.31 (334.36, 8956.71) |
| Day 7 | 2400.49 (1384.87, 5397.56) | 388.13 (118.52, 883.67)a | |
| Day 14 | 2191.53 (375.93, 6433.65) | 143.71 (113.41, 690.34)ab | |
Table 2 Changes of IL-1β and IL-18 contents in wound secretion of two groups of patients [pg/mL, M (Q1, Q3)]
| Item | Control group (n = 10) | HYSJ group (n = 10) | |
|---|---|---|---|
| IL-1β | Day 0 | 2472.35 (379.63, 82825.95) | 2366.69 (1011.29, 15433.128) |
| Day 7 | 1864.33 (994.49, 2583.92) | 604.83 (367.05, 1169.90)a | |
| Day 14 | 2035.07 (875.39, 2933.65) | 548.94 (115.37, 692.52)ab | |
| IL-18 | Day 0 | 1073.13 (412.47, 4554.46) | 2036.31 (334.36, 8956.71) |
| Day 7 | 2400.49 (1384.87, 5397.56) | 388.13 (118.52, 883.67)a | |
| Day 14 | 2191.53 (375.93, 6433.65) | 143.71 (113.41, 690.34)ab | |
Table 3 Changes of VEGF-C content in wound exudates of two groups ($ \bar{x} \pm s$, ng/mL)
| Treatment time | Control group (n = 10) | HYSJ group (n = 10) |
|---|---|---|
| Day 0 | 0.39±0.17 | 0.21±0.31 |
| Day 7 | 0.15±0.10 | 0.45±0.18ab |
| Day 14 | 0.26±0.11 | 0.42±0.19b |
Table 3 Changes of VEGF-C content in wound exudates of two groups ($ \bar{x} \pm s$, ng/mL)
| Treatment time | Control group (n = 10) | HYSJ group (n = 10) |
|---|---|---|
| Day 0 | 0.39±0.17 | 0.21±0.31 |
| Day 7 | 0.15±0.10 | 0.45±0.18ab |
| Day 14 | 0.26±0.11 | 0.42±0.19b |
Figure 3 Analysis of IHC staining of lymphatic vessel formation-related proteins in wound tissues in each group at different intervention time A: IHC staining images of LYVE-1, PROX-1, and VEGFR-3 proteins in the wound tissue of the control group on days 0, 7, and 14 of treatment; B: IHC staining images of LYVE-1, PROX-1, and VEGFR-3 proteins in the wound tissue of the HYSJ group on days 0, 7, and 14 of treatment; C: quantitative IHC analysis of LYVE -1of wound tissue in the two groups; D: quantitative IHC analysis of PROX-1 of wound tissue in the two groups; E: quantitative IHC analysis of VEGFR-3 of wound tissue in the two groups; A1, B1: LYVE-1 protein before intervention (× 40); A2, B2: PROX-1 protein before intervention (× 40); A3, B3: VEGFR-3 protein before intervention (× 40); A4, B4: LYVE-1 on the 7th day of intervention (× 40); A5, B5: PROX-1 on the 7th day of intervention (× 40); A6, B6: VEGFR-3 on the 7th day of intervention (× 40); A7, B7: LYVE-1 on the 14th day of intervention (× 40); A8, B8: PROX-1 on the 14th day of intervention (× 40); A9, B9: VEGFR-3 on the 14th day of intervention (× 40). The arrows indicate positive staining points. Scale bar: 80 μm. HYSJ group: treated with Huiyang Shengji unguent; control group: treated with bFGF. IHC: immunohistochemical; LYVE-1: lymphatic vessel endothelial hyaluronan receptor 1; PROX-1: prospero homeobox protein 1; VEGFR-3: vascular endothelial growth factor receptor 3; HYSJ: Huiyang Shengji unguent; bFGF: basic fibroblast growth factor. Data were expressed as mean ± standard deviation (n = 6). The independent sample t-test was used for between-group comparisons. Compared with control group, aP < 0.001; bP < 0.01; cP < 0.05.
Figure 3 Analysis of IHC staining of lymphatic vessel formation-related proteins in wound tissues in each group at different intervention time A: IHC staining images of LYVE-1, PROX-1, and VEGFR-3 proteins in the wound tissue of the control group on days 0, 7, and 14 of treatment; B: IHC staining images of LYVE-1, PROX-1, and VEGFR-3 proteins in the wound tissue of the HYSJ group on days 0, 7, and 14 of treatment; C: quantitative IHC analysis of LYVE -1of wound tissue in the two groups; D: quantitative IHC analysis of PROX-1 of wound tissue in the two groups; E: quantitative IHC analysis of VEGFR-3 of wound tissue in the two groups; A1, B1: LYVE-1 protein before intervention (× 40); A2, B2: PROX-1 protein before intervention (× 40); A3, B3: VEGFR-3 protein before intervention (× 40); A4, B4: LYVE-1 on the 7th day of intervention (× 40); A5, B5: PROX-1 on the 7th day of intervention (× 40); A6, B6: VEGFR-3 on the 7th day of intervention (× 40); A7, B7: LYVE-1 on the 14th day of intervention (× 40); A8, B8: PROX-1 on the 14th day of intervention (× 40); A9, B9: VEGFR-3 on the 14th day of intervention (× 40). The arrows indicate positive staining points. Scale bar: 80 μm. HYSJ group: treated with Huiyang Shengji unguent; control group: treated with bFGF. IHC: immunohistochemical; LYVE-1: lymphatic vessel endothelial hyaluronan receptor 1; PROX-1: prospero homeobox protein 1; VEGFR-3: vascular endothelial growth factor receptor 3; HYSJ: Huiyang Shengji unguent; bFGF: basic fibroblast growth factor. Data were expressed as mean ± standard deviation (n = 6). The independent sample t-test was used for between-group comparisons. Compared with control group, aP < 0.001; bP < 0.01; cP < 0.05.
Figure 4 Western blot analysis of inflammatory-related proteins in the wound tissues in each group at different intervention time A: relative expression levels of the various proteins in wound tissues in the two patient groups before treatment; B: relative expression levels of the various proteins in wound tissues in the two patient groups of treatment for 7 d; C: relative expression levels of the various proteins in wound tissues in the two patient groups of treatment for 14 d; A1, B1, C1: the protein electrophoresis bands; A2, B2, C2: expression levels of Gasdermin D protein; A3, B3, C3: expression levels of N-Gasdermin D protein; A4, B4, C4: expression levels of caspase-1 protein; A5, B5, C5: expression levels of NLRP3 protein. HYSJ group: treated with Huiyang Shengji unguent; control group: treated with bFGF. NLRP3: NOD-like receptor family, pyrin domain containing protein 3; HYSJ: Huiyang Shengji unguent; bFGF: basic fibroblast growth factor. Data were expressed as mean ± standard deviation (n = 3). The independent sample t-test was used for between-group comparisons. Compared with control group, aP < 0.05, bP < 0.01.
Figure 4 Western blot analysis of inflammatory-related proteins in the wound tissues in each group at different intervention time A: relative expression levels of the various proteins in wound tissues in the two patient groups before treatment; B: relative expression levels of the various proteins in wound tissues in the two patient groups of treatment for 7 d; C: relative expression levels of the various proteins in wound tissues in the two patient groups of treatment for 14 d; A1, B1, C1: the protein electrophoresis bands; A2, B2, C2: expression levels of Gasdermin D protein; A3, B3, C3: expression levels of N-Gasdermin D protein; A4, B4, C4: expression levels of caspase-1 protein; A5, B5, C5: expression levels of NLRP3 protein. HYSJ group: treated with Huiyang Shengji unguent; control group: treated with bFGF. NLRP3: NOD-like receptor family, pyrin domain containing protein 3; HYSJ: Huiyang Shengji unguent; bFGF: basic fibroblast growth factor. Data were expressed as mean ± standard deviation (n = 3). The independent sample t-test was used for between-group comparisons. Compared with control group, aP < 0.05, bP < 0.01.
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