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To investigate the effect of inhibition of mitochondrial pyruvate carrier on Okada-induced cuprotosis in SH-SY5Y cells.
SH-SY5Y cells were induced by okada acid (OA) to establish a model of Alzheimer’s disease (AD). The cells were divided into control group (complete medium), model group (20 nmol·L-1 OA) and experimental group [20 nmol·L-1 OA+1 μmol·L-1 2-cyano-3 -(1-phenyl-1H-indole-3-yl) -2-acrylic acid (UK-5099)]. Copper ion content was detected by cell cupric colorimetric kit. Pyruvate content was detect by pyruvate detection kit, and the expressions of phosphorylated Tau (AT8) and ferredoxin 1 (FDX1) in each group were detected by immunofluorescence staining. The expression levels of mitochondrial pyruvate carrier (MPC1/MPC2) and dihydrolipoyl transacetylase (DLAT) were detected by Western blot.
The contents of copper ions in experimental group, model group and control group were (0.36±0.01), (0.90±0.05) and (0.23±0.04) μmol·g prot-1; the pyruvate content were (0.55±0.04), (0.33±0.03) and (0.44±0.04) μmol·108 cell-1; the AT8 expression levels were 91.46±14.53, 125.02±14.43 and 72.57±19.81; the FDX1 expression levels were 98.93±7.40, 147.83±18.63 and 79.18±8.06; the relative expression levels of MPC1 protein were 0.59±0.11, 1.01±0.14 and 0.55±0.13; the relative expression levels of MPC2 protein were 0.54±0.05, 1.31±0.07 and 0.43±0.07; the relative expression levels of DLAT protein were 0.72±0.04, 0.92±0.07 and 0.53±0.14, respectively. The above indexes in the experimental group were statistically significant compared with those in the model group (P<0.05, P<0.01, P<0.001).
Inhibition of mitochondrial pyruvate carrier can alleviate Okada-induced cuprotosis of SH-SY5Y cells.
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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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