SRS16-86

Cell lines

HT-1080 and NIH 3T3 cells

Preparation Method

Human HT-1080 fibrosarcoma cells were cultured in DMEM supplemented with MEM NEAA, 10% (vol/vol) FCS, 100U/mL penicillin, and 100μg/mL streptomycin. Murine NIH 3T3 cells were cultured in DMEM supplemented with 10% (vol/vol) FCS, 100U/mL penicillin, and 100μg/mL streptomycin. All cell lines were cultured in a humidified 5% CO₂ atmosphere. For induction of ferroptosis, NIH 3T3 and HT-1080 cells were each stimulated for 24h at 37°C with 50μM erastin in the presence or absence of 1μM SRS16-86 (vehicle-treated cells served as control). Then cells were collected for Western Blot analysis. Phosphatidylserine exposure to the outer cell membrane of apoptotic cells or at the inner plasma membrane of necrotic cells and incorporation of 7-AAD into necrotic cells were quantified by fluorescence-activated cell sorting (FACS) analysis.

Reaction Conditions

1μM; 24h

Applications

SRS16-86 completely prevented erastin-induced ferroptosis in HT-1080 and NIH 3T3 cells, abolished 7-AAD/Annexin V positivity cells without activating caspase-3.

Animal models

Sprague–Dawley rats

Preparation Method

Sprague–Dawley rats were intraperitoneally injected with 60mg/kg freshly prepared streptozotocin (STZ) once to cause diabetes, and normal control animals were intraperitoneally injected with the same amount of citric acid buffer. Tail vein blood glucose levels were measured to confirm DM; a random blood glucose level of≥16.7mM was used as the standard for the DM model. The dose of SRS16-86 was 15mg/kg/day from week 1 to week 8 after STZ treatment, because this dose of SRS16-86 was safe and effective for repairing spinal cord injury in the animal model. Six animals from each group at each time point were selected six animals from each group at each time point to ensure the accuracy of the data. Animals were randomly assigned to the following groups: control group, Sprague–Dawley rats+citrate buffer; Diabetic Nephropathy (DN) group, Sprague–Dawley rats+streptozotocin(STZ); and DN-SRS group, Sprague–Dawley rats+STZ+SRS16-86. Twenty-four hour urine samples were collected from Sprague–Dawley rats in each group using metabolic cages at 8, 12 and 16 weeks. Urinary biochemistry was detected by a Urinary Microalbumin Test. After experiment, kidneys were collected for Western blot and Hematoxylin and eosin (HE) staining.

Dosage form

15mg/kg/day; i.p.; 8 week

Applications

SRS16-86 reversed streptozotocin-induced diabetic nephropathy in male Sprague–Dawley rats, reduced 24h urinary total protein and serum creatinine, and decreased renal 4-HNE, MDA, IL-1β and TNF-α while increased GPX4, xCT and GSH levels.

References:
[1] Linkermann A, Skouta R, Himmerkus N, et al. Synchronized renal tubular cell death involves ferroptosis. Proc Natl Acad Sci U S A. 2014;111(47):16836-16841.
[2] Qiao Y, Sun C, Kan S, et al. SRS 16-86 promotes diabetic nephropathy recovery by regulating ferroptosis. Exp Physiol. 2024;109(7):1199-1210.