Limits...
Expression of PHB2 in rat brain cortex following traumatic brain injury.

Xu T, Fan X, Tan Y, Yue Y, Chen W, Gu X - Int J Mol Sci (2014)

Bottom Line: Knocking down PHB2 by siRNA partly increased the apoptosis level of PC12 stimulated by H2O2.While the PHB2 was interrupted by siRNA, the proliferation level of primary cultured astrocytes was inhibited notably than that in the control group.Together with our data, we hypothesized that PHB2 might play an important role in CNS pathophysiology after TBI.

View Article: PubMed Central - PubMed

Affiliation: The Center Laboratory of Huai'an First People's Hospital Nanjing Medical University, Huai'an 223300, China. xuting-1988@163.com.

ABSTRACT
Prohibitin2 (PHB2) is a ubiquitous, evolutionarily strongly conserved protein. It is one of the components of the prohibitin complex, which comprises two highly homologous subunits, PHB1 and PHB2. PHB2 is present in various cellular compartments including the nucleus and mitochondria. Recent studies have identified PHB2 as a multifunctional protein that controls cell proliferation, apoptosis, cristae morphogenesis and the functional integrity of mitochondria. However its distribution and function in the central nervous system (CNS) are not well understood. In this study, we examined PHB2 expression and cellular localization in rats after acute traumatic brain injury (TBI). Western Blot analysis showed PHB2 level was significantly enhanced at five days after injury compared to control, and then declined during the following days. The protein expression of PHB2 was further analyzed by immunohistochemistry. In comparison to contralateral cerebral cortex, we observed a highly significant accumulation of PHB2 at the ipsilateral brain. Immunofluorescence double-labeling showed that PHB2 was co-expressed with NeuN, GFAP. Besides, PHB2 also colocalized with activated caspase-3 and PCNA. To further investigate the function of PHB2, primary cultured astrocytes and the neuronal cell line PC12 were employed to establish a proliferation model and an apoptosis model, respectively, to simulate the cell activity after TBI to a certain degree. Knocking down PHB2 by siRNA partly increased the apoptosis level of PC12 stimulated by H2O2. While the PHB2 was interrupted by siRNA, the proliferation level of primary cultured astrocytes was inhibited notably than that in the control group. Together with our data, we hypothesized that PHB2 might play an important role in CNS pathophysiology after TBI.

Show MeSH

Related in: MedlinePlus

The expression change of PHB2 in H2O2-induced PC12 cells apoptosis. (A) The time courses of PHB2 expressions in PC12 cells after H2O2 stimulus; β-actin was used to confirm equal amounts of protein run on gel; (B) Quantification was made at each time point, the bar chart showed the ratio of PHB2 to β-actin at each time point; these data are means ± SEM. * p < 0.05, ** p < 0.01 (n = 3).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC3958913&req=5

f7-ijms-15-03299: The expression change of PHB2 in H2O2-induced PC12 cells apoptosis. (A) The time courses of PHB2 expressions in PC12 cells after H2O2 stimulus; β-actin was used to confirm equal amounts of protein run on gel; (B) Quantification was made at each time point, the bar chart showed the ratio of PHB2 to β-actin at each time point; these data are means ± SEM. * p < 0.05, ** p < 0.01 (n = 3).

Mentions: Previous studies showed PHB2 colocalization with active caspase-3 in neurons, thus we suspected PHB2 might take part in the process of neuronal apoptosis after TBI. We explored a H2O2-induced PC12 cell apoptosis model for subsequent research. A certain stimulation concentration of H2O2 (0.25 μM) was added to the PC12 cells. Cell protein was collected at 4, 6, 8, 10, 12, and 24 h after stimulation. Western Blot assays were used to detect the expression levels of PHB2 at each time point. Results showed that PHB2 increased in PC12 stimulated by H2O2, and peaked at 8 h (Figure 7A). These results indicate that PHB2 might play a role in apoptosis of PC12 cells stimulated by H2O2. However, further study is required to determine on whether it acts as protected or not.


Expression of PHB2 in rat brain cortex following traumatic brain injury.

Xu T, Fan X, Tan Y, Yue Y, Chen W, Gu X - Int J Mol Sci (2014)

The expression change of PHB2 in H2O2-induced PC12 cells apoptosis. (A) The time courses of PHB2 expressions in PC12 cells after H2O2 stimulus; β-actin was used to confirm equal amounts of protein run on gel; (B) Quantification was made at each time point, the bar chart showed the ratio of PHB2 to β-actin at each time point; these data are means ± SEM. * p < 0.05, ** p < 0.01 (n = 3).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3958913&req=5

f7-ijms-15-03299: The expression change of PHB2 in H2O2-induced PC12 cells apoptosis. (A) The time courses of PHB2 expressions in PC12 cells after H2O2 stimulus; β-actin was used to confirm equal amounts of protein run on gel; (B) Quantification was made at each time point, the bar chart showed the ratio of PHB2 to β-actin at each time point; these data are means ± SEM. * p < 0.05, ** p < 0.01 (n = 3).
Mentions: Previous studies showed PHB2 colocalization with active caspase-3 in neurons, thus we suspected PHB2 might take part in the process of neuronal apoptosis after TBI. We explored a H2O2-induced PC12 cell apoptosis model for subsequent research. A certain stimulation concentration of H2O2 (0.25 μM) was added to the PC12 cells. Cell protein was collected at 4, 6, 8, 10, 12, and 24 h after stimulation. Western Blot assays were used to detect the expression levels of PHB2 at each time point. Results showed that PHB2 increased in PC12 stimulated by H2O2, and peaked at 8 h (Figure 7A). These results indicate that PHB2 might play a role in apoptosis of PC12 cells stimulated by H2O2. However, further study is required to determine on whether it acts as protected or not.

Bottom Line: Knocking down PHB2 by siRNA partly increased the apoptosis level of PC12 stimulated by H2O2.While the PHB2 was interrupted by siRNA, the proliferation level of primary cultured astrocytes was inhibited notably than that in the control group.Together with our data, we hypothesized that PHB2 might play an important role in CNS pathophysiology after TBI.

View Article: PubMed Central - PubMed

Affiliation: The Center Laboratory of Huai'an First People's Hospital Nanjing Medical University, Huai'an 223300, China. xuting-1988@163.com.

ABSTRACT
Prohibitin2 (PHB2) is a ubiquitous, evolutionarily strongly conserved protein. It is one of the components of the prohibitin complex, which comprises two highly homologous subunits, PHB1 and PHB2. PHB2 is present in various cellular compartments including the nucleus and mitochondria. Recent studies have identified PHB2 as a multifunctional protein that controls cell proliferation, apoptosis, cristae morphogenesis and the functional integrity of mitochondria. However its distribution and function in the central nervous system (CNS) are not well understood. In this study, we examined PHB2 expression and cellular localization in rats after acute traumatic brain injury (TBI). Western Blot analysis showed PHB2 level was significantly enhanced at five days after injury compared to control, and then declined during the following days. The protein expression of PHB2 was further analyzed by immunohistochemistry. In comparison to contralateral cerebral cortex, we observed a highly significant accumulation of PHB2 at the ipsilateral brain. Immunofluorescence double-labeling showed that PHB2 was co-expressed with NeuN, GFAP. Besides, PHB2 also colocalized with activated caspase-3 and PCNA. To further investigate the function of PHB2, primary cultured astrocytes and the neuronal cell line PC12 were employed to establish a proliferation model and an apoptosis model, respectively, to simulate the cell activity after TBI to a certain degree. Knocking down PHB2 by siRNA partly increased the apoptosis level of PC12 stimulated by H2O2. While the PHB2 was interrupted by siRNA, the proliferation level of primary cultured astrocytes was inhibited notably than that in the control group. Together with our data, we hypothesized that PHB2 might play an important role in CNS pathophysiology after TBI.

Show MeSH
Related in: MedlinePlus