Cytochrome C (from pig)

Cell experiment [1]:

Cell lines

THP-1 cells and HEK 293T cell

Preparation method

Cell lysates were prepared from LPS stimulated differentiated THP-1 cells and HEK 293T cell expressing HA-NEK7 or not. The cell lysates were mixed and incubated with or without cytochrome c (0.25 mg/ml) for 30 minutes at 30°C. The status of full length and cleaved caspase-9 was assessed by immunoblotting.

Reaction Conditions

0.25 mg/ml; 30 minutes at 30°C

Applications

In the absence of cytochrome c an increase in cleaved caspase-1 in NEK7 overexpressed cells, while co-expression of cytochrome c inhibited the increase.

Animal experiment [2]:

Animal models

Sprague-Dawley rats

Preparation method

Thirty Sprague-Dawley rats were randomly divided into the vehicle control group, PMB group, PMB + cyto c (Cytochrome c) 50, 100, or 200 mg/kg group, respectively, and were treated with intravenous cyto c 30 min before the administration of PMB 4.0 mg/kg once a day for consecutive 5 days. On the 4th day after administration, 24 h urine was collected to determine N-acetyl-β-D-glucosaminidase excretion. Six hours after the last injection on the 5th day, kidneys were harvested to assay PMB concentration and observe pathological alterations, and blood samples were collected to assay serum creatinine (SCr), blood urea nitrogen (BUN), and blood β2-microglobulin (β2-MG) levels.

Dosage form

50, 100, or 200 mg/kg; i.v.

Applications

Cyto c 50, 100, and 200 mg/kg decreased the accumulation of PMB in the kidney by 18.5%, 39.1%, and 36.8%, respectively, and reduced 24 h N-acetyl-β-D- glucosaminidase excretion by 22.5% , 40.4% , and 40.4%, respectively. Kidney pathological damage induced by PMB was markedly reduced by cyto c 100 mg/kg and 200 mg/kg.

References:

[1] Shi CS, Kehrl JH. Cytochrome c Negatively Regulates NLRP3 Inflammasomes. PLoS One. 2016 Dec 28;11(12):e0167636.

[2] Li ZD, et al. Cytochrome C suppresses renal accumulation and nephrotoxicity of polymyxin B. Hum Exp Toxicol. 2019 Feb;38(2):193-200.