Journal of Traditional Chinese Medicine ›› 2026, Vol. 46 ›› Issue (5): 1160-1171.DOI: 10.19852/j.cnki.jtcm.2026.05.010
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Exploring the role of Yishen Qufeng formula (益肾祛风方) in alleviating renal injury in diabetic kidney disease rats via regulating pyroptosis through the nucleotide-binding oligomerization domain-like receptor protein 3/caspase-1/gasdermin D pathway
ZHAO Xueshan1,2,3, CHEN Ansheng1,2,3, YONG Chen2,3, HUANG Guoshun1,2,3, ZHOU Enchao1,2,3(
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- 1
the First Clinical Medical College ,Nanjing University of Chinese Medicine Nanjing 210023, China
2Department of Nephrology ,Affiliated Hospital of Nanjing University of Chinese Medicine Nanjing 210029, China
3Jiangsu Key Laboratory of Traditional Chinese Medicine for Kidney Tonification and Anti-Aging Nanjing 210023, China
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Received:2025-08-20Accepted:2025-12-03Online:2026-10-15Published:2026-10-09 -
Contact:Prof. ZHOU Enchao, Nanjing University of Chinese Medicine, Nanjing 210023, China; Department of Nephrology, Affiliated Hospital of Nanjing University of Chinese Medicine, Nanjing 210029, China; Jiangsu Key Laboratory of Traditional Chinese Medicine for Kidney Tonification and Anti-Aging, Nanjing 210023, China. zhouenchao@njucm.edu.cn,Telephone: +86-13851572603 -
Supported by:National Natural Science Foundation of China: Mechanism Study of Yishen Qufeng Formula in Regulating the A-Kinase Anchoring Protein 1-mtDNA Pathway to Inhibit Z-DNA Binding Protein 1 (ZBP1)-Mediated PANoptosis of Renal Tubular Epithelial Cells and Ameliorate Diabetic Kidney Disease(82474427);Third Batch of Jiangsu Provincial Leading Talent Cultivation Program in Traditional Chinese Medicine: Evidence-Based Protocol for the Integrated Prevention and Treatment of Chronic Kidney Disease via Tonifying Kidney Primordial-Qi and Eliminating Renal Toxins and its Mechanistic Research(013091001009);Postgraduate Research & Practice Innovation Program of Jiangsu Province: Clinical Efficacy and Mechanism of Yishen Qufeng Formula for Diabetic Kidney Disease: Cyclic Guanosine Monophosphate-Adenosine Monophosphate Synthase/Stimulator of Interferon Genes Axis Induced by Coordinated Sensing of ZBP1 and mtDNA(SJCX25_0939)
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ZHAO Xueshan, CHEN Ansheng, YONG Chen, HUANG Guoshun, ZHOU Enchao. Exploring the role of Yishen Qufeng formula (益肾祛风方) in alleviating renal injury in diabetic kidney disease rats via regulating pyroptosis through the nucleotide-binding oligomerization domain-like receptor protein 3/caspase-1/gasdermin D pathway[J]. Journal of Traditional Chinese Medicine, 2026, 46(5): 1160-1171.
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Figure 1 General condition of rats A: general appearance of rats in each group; A1: control group; A2: model group; A3: IRB group; A4: YSQF group; B: changes in body weight over time in each group; C: gross renal morphology; C1: control group; C2: model group; C3: IRB group; C4: YSQF group; D: KW/BW in each group. Control: control group (sham surgery without kidney removal + intraperitoneal injections of sodium citrate buffer + sterile water); Model: model group (unilateral nephrectomy + intraperitoneal injections of STZ + sterile water); IRB: IRB group (unilateral nephrectomy + intraperitoneal injections of STZ + 13.5 mg/kg IRB); YSQF: YSQF group (unilateral nephrectomy + intraperitoneal injections of STZ + 13.86 g/kg YSQF). IRB: irbesartan; YSQF: Yishen Qufeng formula; STZ: streptozotocin; KW/BW: kidney weight/body weight ratio. Body weight data at weeks 1, 5, 9, and 17 followed a normal distribution and were analyzed using one-way analysis of variance, whereas data at week 13 were non-normally distributed and were analyzed using a non-parametric test. For visual consistency, all body weight data in the line graph are presented as mean ± standard deviation: n = 8 for the control group, n = 9 for the model group, n = 7 for the IRB group, and n = 7 for the YSQF group. aP < 0.05, bP < 0.01, cP < 0.001, compared with the control group; dP < 0.001, compared with the model group; eP < 0.05, compared with the IRB group.
Figure 1 General condition of rats A: general appearance of rats in each group; A1: control group; A2: model group; A3: IRB group; A4: YSQF group; B: changes in body weight over time in each group; C: gross renal morphology; C1: control group; C2: model group; C3: IRB group; C4: YSQF group; D: KW/BW in each group. Control: control group (sham surgery without kidney removal + intraperitoneal injections of sodium citrate buffer + sterile water); Model: model group (unilateral nephrectomy + intraperitoneal injections of STZ + sterile water); IRB: IRB group (unilateral nephrectomy + intraperitoneal injections of STZ + 13.5 mg/kg IRB); YSQF: YSQF group (unilateral nephrectomy + intraperitoneal injections of STZ + 13.86 g/kg YSQF). IRB: irbesartan; YSQF: Yishen Qufeng formula; STZ: streptozotocin; KW/BW: kidney weight/body weight ratio. Body weight data at weeks 1, 5, 9, and 17 followed a normal distribution and were analyzed using one-way analysis of variance, whereas data at week 13 were non-normally distributed and were analyzed using a non-parametric test. For visual consistency, all body weight data in the line graph are presented as mean ± standard deviation: n = 8 for the control group, n = 9 for the model group, n = 7 for the IRB group, and n = 7 for the YSQF group. aP < 0.05, bP < 0.01, cP < 0.001, compared with the control group; dP < 0.001, compared with the model group; eP < 0.05, compared with the IRB group.
Table 1 Biochemical parameters of blood and urine in rats
| Group | n | Scr (µmol/L) | BUN (mmol/L) | K (mmol/L) | ALT (U/L) | AST (U/L) | 24-h UTP (mg/24 h) | ACR (mg/g) | |||
|---|---|---|---|---|---|---|---|---|---|---|---|
| Control | 8 | 27.9±2.2 | 3.6±0.7 | 7.5 (6.1, 7.9) | 51.0 (44.5, 53.5) | 99.5 (84.8, 108.0) | 10.4 (5.8, 17.6) | 740.0 (370.0, 1903.0) | |||
| Model | 9 | 32.8±13.1 | 9.7±1.8b | 8.3 (7.8, 9.0)a | 82.0 (65.0, 613.5)a | 122.0 (101.5, 770.5) | 44.9 (38.3, 60.7)b | 7211.0 (3086.5, 13433.5)b | |||
| IRB | 7 | 23.2±2.7a | 8.9±1.2b | 10.6 (10.1, 12.9)bc | 106.0 (69.0, 160.0)a | 195.0 (77.0, 413.0) | 27.4 (11.5, 37.2)ae | 3697.0 (3084.0, 4845.0)b | |||
| YSQF | 7 | 23.6±6.4 | 8.9±1.1b | 10.5 (9.4, 11.0)bd | 80.0 (65.0, 82.0) | 134.0 (106.0, 184.0) | 18.9 (14.5, 40.9)ad | 3862.0 (2773.0, 5066.0)b | |||
Table 1 Biochemical parameters of blood and urine in rats
| Group | n | Scr (µmol/L) | BUN (mmol/L) | K (mmol/L) | ALT (U/L) | AST (U/L) | 24-h UTP (mg/24 h) | ACR (mg/g) | |||
|---|---|---|---|---|---|---|---|---|---|---|---|
| Control | 8 | 27.9±2.2 | 3.6±0.7 | 7.5 (6.1, 7.9) | 51.0 (44.5, 53.5) | 99.5 (84.8, 108.0) | 10.4 (5.8, 17.6) | 740.0 (370.0, 1903.0) | |||
| Model | 9 | 32.8±13.1 | 9.7±1.8b | 8.3 (7.8, 9.0)a | 82.0 (65.0, 613.5)a | 122.0 (101.5, 770.5) | 44.9 (38.3, 60.7)b | 7211.0 (3086.5, 13433.5)b | |||
| IRB | 7 | 23.2±2.7a | 8.9±1.2b | 10.6 (10.1, 12.9)bc | 106.0 (69.0, 160.0)a | 195.0 (77.0, 413.0) | 27.4 (11.5, 37.2)ae | 3697.0 (3084.0, 4845.0)b | |||
| YSQF | 7 | 23.6±6.4 | 8.9±1.1b | 10.5 (9.4, 11.0)bd | 80.0 (65.0, 82.0) | 134.0 (106.0, 184.0) | 18.9 (14.5, 40.9)ad | 3862.0 (2773.0, 5066.0)b | |||
Table 2 Serum inflammatory cytokine levels in rats (ng/L, $\bar{x}$ ± s)
| Group | n | IL-1β | IL-18 |
|---|---|---|---|
| Control | 8 | 35.8±2.1 | 168.4±10.1 |
| Model | 9 | 45.7±1.6a | 210.5±7.2a |
| IRB | 7 | 37.5±1.5b | 186.6±4.8db |
| YSQF | 7 | 41.4±1.9abc | 196.0±4.7aef |
Table 2 Serum inflammatory cytokine levels in rats (ng/L, $\bar{x}$ ± s)
| Group | n | IL-1β | IL-18 |
|---|---|---|---|
| Control | 8 | 35.8±2.1 | 168.4±10.1 |
| Model | 9 | 45.7±1.6a | 210.5±7.2a |
| IRB | 7 | 37.5±1.5b | 186.6±4.8db |
| YSQF | 7 | 41.4±1.9abc | 196.0±4.7aef |
Figure 2 Histopathological changes of renal tissues of rats A: HE staining (× 20, scale bar = 50 μm); B: Masson’s staining (× 20, scale bar = 50 μm); C: TUNEL staining (× 20, scale bar = 50 μm); A1, B1, C1: control group glomeruli; A2, B2, C2: model group glomeruli; A3, B3, C3: IRB group glomeruli; A4, B4, C4: YSQF group glomeruli; A5, B5, C5: control group renal tubules; A6, B6, C6: model group renal tubules; A7, B7, C7: IRB group renal tubules; A8, B8, C8: YSQF group renal tubules. D: area percentage of renal collagen content; E: apoptotic positive area percentage in renal tissue. Control: control group (sham surgery without kidney removal + intraperitoneal injections of sodium citrate buffer + sterile water); Model: model group (unilateral nephrectomy + intraperitoneal injections of STZ + sterile water); IRB: IRB group (unilateral nephrectomy + intraperitoneal injections of STZ + 13.5 mg/kg IRB); YSQF: YSQF group (unilateral nephrectomy + intraperitoneal injections of STZ + 13.86 g/kg YSQF). IRB: irbesartan; YSQF: Yishen Qufeng formula; STZ: streptozotocin; HE: hematoxylin and eosin staining; Masson’s staining: Masson's trichrome staining; TUNEL staining: terminal deoxynucleotidyl transferase dUTP nick end labeling staining. Normally distributed data were expressed as mean ± standard deviation and compared using one-way analysis of variance (n = 3). aP < 0.01, dP < 0.001, eP < 0.05, compared with the control group; bP < 0.05, cP < 0.01, compared with the model group.
Figure 2 Histopathological changes of renal tissues of rats A: HE staining (× 20, scale bar = 50 μm); B: Masson’s staining (× 20, scale bar = 50 μm); C: TUNEL staining (× 20, scale bar = 50 μm); A1, B1, C1: control group glomeruli; A2, B2, C2: model group glomeruli; A3, B3, C3: IRB group glomeruli; A4, B4, C4: YSQF group glomeruli; A5, B5, C5: control group renal tubules; A6, B6, C6: model group renal tubules; A7, B7, C7: IRB group renal tubules; A8, B8, C8: YSQF group renal tubules. D: area percentage of renal collagen content; E: apoptotic positive area percentage in renal tissue. Control: control group (sham surgery without kidney removal + intraperitoneal injections of sodium citrate buffer + sterile water); Model: model group (unilateral nephrectomy + intraperitoneal injections of STZ + sterile water); IRB: IRB group (unilateral nephrectomy + intraperitoneal injections of STZ + 13.5 mg/kg IRB); YSQF: YSQF group (unilateral nephrectomy + intraperitoneal injections of STZ + 13.86 g/kg YSQF). IRB: irbesartan; YSQF: Yishen Qufeng formula; STZ: streptozotocin; HE: hematoxylin and eosin staining; Masson’s staining: Masson's trichrome staining; TUNEL staining: terminal deoxynucleotidyl transferase dUTP nick end labeling staining. Normally distributed data were expressed as mean ± standard deviation and compared using one-way analysis of variance (n = 3). aP < 0.01, dP < 0.001, eP < 0.05, compared with the control group; bP < 0.05, cP < 0.01, compared with the model group.
Figure 3 Changes in pyroptosis-related proteins in renal tissue of rats A: relative mRNA expression of NLRP3; B: relative mRNA expression of ASC; C: electrophoresis band pattern; D: relative protein expression levels of NLRP3; E: relative protein expression levels of ASC; F: relative protein expression levels of cleaved-caspase-1; G: relative protein expression levels of GSDMD-N. Control: control group (sham surgery without kidney removal + intraperitoneal injections of sodium citrate buffer + sterile water); Model: model group (unilateral nephrectomy + intraperitoneal injections of STZ + sterile water); IRB: IRB group (unilateral nephrectomy + intraperitoneal injections of STZ + 13.5 mg/kg IRB); YSQF: YSQF group (unilateral nephrectomy + intraperitoneal injections of STZ + 13.86 g/kg YSQF). IRB: irbesartan; YSQF: Yishen Qufeng formula; STZ: streptozotocin; NLRP3: nucleotide-binding oligomerization domain-like receptor protein 3; ASC: apoptosis-associated speck-like protein containing a caspase recruitment domain; cleaved-caspase-1: cleaved cysteine-aspartic acid protease 1; GSDMD-N: N-terminal fragment of gasdermin D. Normally distributed data were expressed as mean ± standard deviation and compared using one-way analysis of variance (n = 3). aP < 0.001, bP < 0.01, eP < 0.05, compared with the control group; cP < 0.001, gP < 0.05, compared with the model group, dP < 0.01, fP < 0.05, compared with the control group.
Figure 3 Changes in pyroptosis-related proteins in renal tissue of rats A: relative mRNA expression of NLRP3; B: relative mRNA expression of ASC; C: electrophoresis band pattern; D: relative protein expression levels of NLRP3; E: relative protein expression levels of ASC; F: relative protein expression levels of cleaved-caspase-1; G: relative protein expression levels of GSDMD-N. Control: control group (sham surgery without kidney removal + intraperitoneal injections of sodium citrate buffer + sterile water); Model: model group (unilateral nephrectomy + intraperitoneal injections of STZ + sterile water); IRB: IRB group (unilateral nephrectomy + intraperitoneal injections of STZ + 13.5 mg/kg IRB); YSQF: YSQF group (unilateral nephrectomy + intraperitoneal injections of STZ + 13.86 g/kg YSQF). IRB: irbesartan; YSQF: Yishen Qufeng formula; STZ: streptozotocin; NLRP3: nucleotide-binding oligomerization domain-like receptor protein 3; ASC: apoptosis-associated speck-like protein containing a caspase recruitment domain; cleaved-caspase-1: cleaved cysteine-aspartic acid protease 1; GSDMD-N: N-terminal fragment of gasdermin D. Normally distributed data were expressed as mean ± standard deviation and compared using one-way analysis of variance (n = 3). aP < 0.001, bP < 0.01, eP < 0.05, compared with the control group; cP < 0.001, gP < 0.05, compared with the model group, dP < 0.01, fP < 0.05, compared with the control group.
Figure 4 NLRP3 immunohistochemical expression in renal tissue of rats. A: immunohistochemical staining (× 20, scale bar = 50 μm); A1: control group glomeruli; A2: model group glomeruli; A3: IRB group glomeruli; A4: YSQF group glomeruli; A5: control group renal tubules; A6: model group renal tubules; A7: IRB group renal tubules; A8: YSQF group renal tubules; B: positive area percentage of NLRP3 staining. Control: control group (sham surgery without kidney removal + intraperitoneal injections of sodium citrate buffer + sterile water); Model: model group (unilateral nephrectomy + intraperitoneal injections of STZ + sterile water); IRB: IRB group (unilateral nephrectomy + intraperitoneal injections of STZ + 13.5 mg/kg IRB); YSQF: YSQF group (unilateral nephrectomy + intraperitoneal injections of STZ + 13.86 g/kg YSQF). IRB: irbesartan; YSQF: Yishen Qufeng formula; STZ: streptozotocin; NLRP3: nucleotide-binding oligomerization domain-like receptor protein 3. Normally distributed data were expressed as mean ± standard deviation and compared using one-way analysis of variance (n = 3). aP < 0.001, compared with the control group; bP < 0.01, compared with the model group.
Figure 4 NLRP3 immunohistochemical expression in renal tissue of rats. A: immunohistochemical staining (× 20, scale bar = 50 μm); A1: control group glomeruli; A2: model group glomeruli; A3: IRB group glomeruli; A4: YSQF group glomeruli; A5: control group renal tubules; A6: model group renal tubules; A7: IRB group renal tubules; A8: YSQF group renal tubules; B: positive area percentage of NLRP3 staining. Control: control group (sham surgery without kidney removal + intraperitoneal injections of sodium citrate buffer + sterile water); Model: model group (unilateral nephrectomy + intraperitoneal injections of STZ + sterile water); IRB: IRB group (unilateral nephrectomy + intraperitoneal injections of STZ + 13.5 mg/kg IRB); YSQF: YSQF group (unilateral nephrectomy + intraperitoneal injections of STZ + 13.86 g/kg YSQF). IRB: irbesartan; YSQF: Yishen Qufeng formula; STZ: streptozotocin; NLRP3: nucleotide-binding oligomerization domain-like receptor protein 3. Normally distributed data were expressed as mean ± standard deviation and compared using one-way analysis of variance (n = 3). aP < 0.001, compared with the control group; bP < 0.01, compared with the model group.
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