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To investigate the protective effect and underlying mechanism of kaempferol (Kae) on flap ischemia-reperfusion injury (FIRI) in rats by regulating glutathione peroxidase 4 (GPX4)-mediated ferroptosis and gasdermin D (GSDMD)-mediated pyroptosis.
McFarlane method was used to establish the ischemia-reperfusion model of rat dorsal flap. A total of 91 SPF SD rats were randomly divided into sham group, model group, experimental group, agonist group, inhibitor group, Kae+agonist group and Kae+inhibitor group, with 13 rats in each group. After successful modeling, the experimental group was given kaempferol 50 mg·kg-1 by gavage; the agonist group was intraperitoneally injected with 2 mg·kg-1 NOD like receptor thermoprotein domain associated protein 3 (NLRP3) activator once a week; the inhibitor group was intraperitoneally injected with 30 mg·kg-1 disulfiram once a week; the Kae+agonist group received intraperitoneal injection of 2 mg·kg-1 NLRP3 activator+50 mg·kg-1 kaempferol; the Kae+inhibitor group received intraperitoneal injection of 30 mg·kg-1 disulfiram+50 mg·kg-1 kaempferol for 2 weeks. The iron content in tissues was detected by iron content detection kit; the levels of serum interleukin-1 β (IL-1 β) and IL-18 were detected by enzyme linked immunosorbent assay (ELISA); the expressions of proliferating cell nuclear antigen (Ki67) and platelet endothelial cell adhesion molecule-1 (CD31) were detected by immunohistochemistry; the protein expressions of GPX4, Caspase-1 and GSDMD-NT were detected by Western blot.
The flap survival rates of the sham group, model group, experimental group, agonist group, inhibitor group, Kae+agonist group and Kae+inhibitor group were (96.73±1.10)%, (67.04±1.60)%, (85.39±1.22)%, (57.96±1.52)%, (75.80±1.72)%, (80.28±0.95)% and (90.49±0.76)%, respectively; the tissue iron content levels were (11.57±4.89), (68.04±7.64), (43.87±9.13), (85.97±8.41), (47.78±11.11), (69.68±6.17) and (29.36±7.66) μmol·L-1; the levels of IL-1 β were (43.83±13.96), (246.02±37.60), (172.19±29.46), (354.39±36.05), (153.56±26.89), (271.13±39.54) and (97.53±24.67) pg·mL-1, respectively; the relative expression levels of CD31 were 6 694.96±711.29, 2 126.32±277.21, 3 592.33±317.33, 484.36±56.24, 3 568.83±316.65, 1 901.66±383.85 and 4 804.18±360.03, respectively; the relative expression levels of Ki67 were 75.56±6.57, 24.11±4.02, 37.94±4.08, 12.17±2.28, 38.01±3.80, 24.32±2.76 and 54.65±7.01, respectively; the relative expression levels of GPX4 were 0.92±0.02, 0.30±0.07, 0.47±0.08, 0.12±0.04, 0.47±0.08, 0.27±0.07 and 0.62±0.13, respectively; the relative expression levels of Caspase-1 were 0.08±0.03, 0.40±0.07, 0.29±0.04, 0.68±0.07, 0.28±0.03, 0.40±0.06 and 0.18±0.06, respectively; the relative expression levels of GSDMD were 0.06±0.04, 0.57±0.05, 0.30±0.04, 0.93±0.09, 0.30±0.03, 0.56±0.07 and 0.16±0.02 respectively. There were statistically significant differences in the above indexes between model group and sham group, between experimental group and model group, and between Kae+agonist group, Kae+inhibitor group and experimental group (all P<0.05).
Kaempferol can alleviate flap ischemia-reperfusion injury in rats, and its protective effect may be related to upregulating GPX4 expression, inhibiting ferroptosis, and regulating the NLRP3/Caspase-1/GSDMD pyroptosis signaling pathway.
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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