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Outline
To explore the potential role and underlying mechanism of the functionally uncharacterized gene Gm49394 on regulating β-cell apoptosis under diabetic conditions.
The expression and translational activity of Gm49394 in pancreatic β-cell lines and non-β-cell lines were validated using RNA fluorescence in situ hybridization (RNA-FISH), quantitative real-time PCR (qPCR), Western blotting, and immunofluorescence (IF) assay. The β-cell lines (NIT-1/Min6) with Gm49394 overexpression or knockdown were constructed. The proliferation, apoptosis, mitochondrial function, as well as oxidative stress and endoplasmic reticulum stress markers in these β-cell lines under physiological homeostasis or pathological stress conditions, such as high glucose (30 mmol/L), inflammation (10 ng/mL IFN-γ alone or combined with 10 ng/mL IL-6), and hydrogen peroxide (100 μmol/L H2O2) were detected by flow cytometry and Western blotting.
RNA-FISH and qPCR indicated that Gm49394 was specifically expressed in pancreatic β-cell lines and up-regulated under high glucose or inflammatory stimulation. IF assay and Western blotting showed that Gm49394 had protein-coding activity. Flow cytometry and Western blotting identified that Gm49394 overexpression did not affect β-cell proliferation, but promoted β-cell apoptosis and increased reactive oxygen species (ROS) and mitochondrial superoxide (MitoSOX) levels in β cells under physiological homeostasis or pathological stress conditions (P<0.05). Under physiological conditions, Gm49394 knockdown failed to induce significant alterations on β-cell apoptosis, ROS, or MitoSOX levels. Under pathological stress conditions, Gm49394 knockdown significantly suppressed β-cell proliferation, apoptosis, as well as oxidative and endoplasmic reticulum stress (P<0.05).
Gm49394 may promote β-cell apoptosis via oxidative stress and endoplasmic reticulum stress.
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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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