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Hsp20 Protects against Oxygen-Glucose Deprivation/Reperfusion-Induced Golgi Fragmentation and Apoptosis through Fas/FasL Pathway.

Zhong B, Hu Z, Tan J, Lu T, Lei Q, Chen C, Zeng L - Oxid Med Cell Longev (2015)

Bottom Line: Finding effective neuroprotective agents has become a priority in the treatment of ischemic stroke.However, transfection with Hsp20 significantly attenuates OGDR-induced Golgi fragmentation and apoptosis.Hsp20 interacts with Bax, decreases FasL and Bax expression, and inhibits caspases 3 and p115 cleavage in N2a cells exposed to OGDR.

View Article: PubMed Central - PubMed

Affiliation: Department of Neurology, Second Xiangya Hospital, Central South University, Changsha, Hunan 410011, China ; Department of Traditional Chinese Medicine, Second Xiangya Hospital, Central South University, Changsha, Hunan 410011, China.

ABSTRACT
Cerebral ischemia-reperfusion injury plays an important role in the development of tissue injury after acute ischemic stroke. Finding effective neuroprotective agents has become a priority in the treatment of ischemic stroke. The Golgi apparatus (GA) is a pivotal organelle and its protection is an attractive target in the treatment of cerebral ischemia-reperfusion injury. Protective effects of Hsp20, a potential cytoprotective agent due to its chaperone-like activity and involvement in regulation of many vital processes, on GA were assessed in an ischemia-reperfusion injury model. Mouse neuroblastoma Neuro2a (N2a) cells were subjected to oxygen-glucose deprivation/reperfusion (OGDR) insult. OGDR induces Golgi fragmentation, apoptosis, and p115 cleavage in N2a cells. However, transfection with Hsp20 significantly attenuates OGDR-induced Golgi fragmentation and apoptosis. Hsp20 interacts with Bax, decreases FasL and Bax expression, and inhibits caspases 3 and p115 cleavage in N2a cells exposed to OGDR. Our data demonstrate that increased Hsp20 expression protects against OGDR-induced Golgi fragmentation and apoptosis, likely through interaction with Bax and subsequent amelioration of the OGDR-induced elevation in p115 cleavage via the Fas/FasL signaling pathway. This neuroprotective potential of Hsp20 against OGDR insult and the underlying mechanism will pave the way for its potential clinical application for cerebral ischemia-reperfusion related disorders.

No MeSH data available.


Related in: MedlinePlus

Effects of Hsp20 on GA morphology and apoptosis in N2a cells exposed to OGDR. N2a cells were transfected with different plasmids (pEGFP-N1 and pEGFP-Hsp20). After transfection for 36 h, cells were treated with OGD 4 h plus 12 h reperfusion. The experiment was repeated independently for at least three times. (a) Digital photomicrograph under fluorescent illumination showing the morphology of GA was detected using Golgi58 staining. GA displayed typical ribbon-like structures adjacent to the nuclei in N2a cell transfected with pEGFP-N1 without OGDR exposure. Fragmented GA was evident in N2a cells transfected with pEGFP-N1 exposed to 4 h OGD plus 12 h reperfusion insult. Transfection with Hsp20 significantly attenuated OGDR-induced fragmentation of GA. (b) Quantitation (mean ± SEM) of (a) from three independent experiments. (c) Overexpression of Hsp20 in vitro reduced OGDR-induced apoptosis. Values are expressed as mean ± SEM. ∗P < 0.05 compared to control. ∗∗P < 0.01 compared to control (Bar = 10 μm).
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fig3: Effects of Hsp20 on GA morphology and apoptosis in N2a cells exposed to OGDR. N2a cells were transfected with different plasmids (pEGFP-N1 and pEGFP-Hsp20). After transfection for 36 h, cells were treated with OGD 4 h plus 12 h reperfusion. The experiment was repeated independently for at least three times. (a) Digital photomicrograph under fluorescent illumination showing the morphology of GA was detected using Golgi58 staining. GA displayed typical ribbon-like structures adjacent to the nuclei in N2a cell transfected with pEGFP-N1 without OGDR exposure. Fragmented GA was evident in N2a cells transfected with pEGFP-N1 exposed to 4 h OGD plus 12 h reperfusion insult. Transfection with Hsp20 significantly attenuated OGDR-induced fragmentation of GA. (b) Quantitation (mean ± SEM) of (a) from three independent experiments. (c) Overexpression of Hsp20 in vitro reduced OGDR-induced apoptosis. Values are expressed as mean ± SEM. ∗P < 0.05 compared to control. ∗∗P < 0.01 compared to control (Bar = 10 μm).

Mentions: To further analyze the effects of Hsp20 on OGDR-induced Golgi fragmentation, N2a cells were transfected with different plasmids (pEGFP-N1 and pEGFP-Hsp20). After transfection for 36 h, cells were treated with OGD 4 h plus 12 h reperfusion. Results showed that, in N2a cell transfected with empty vector (pEGFP-N1) without OGDR exposure, GA displayed typical ribbon-like structures adjacent to the nuclei (Figure 3(a)). When N2a cell transfected with pEGFP-N1 were exposed to OGD 4 h plus 12 h reperfusion, lots of GA changed to debris-like structures scattered in the cytoplasm. Meanwhile, transfection with Hsp20 significantly attenuated OGDR-induced Golgi fragmentation (Figures 3(a) and 3(b)).


Hsp20 Protects against Oxygen-Glucose Deprivation/Reperfusion-Induced Golgi Fragmentation and Apoptosis through Fas/FasL Pathway.

Zhong B, Hu Z, Tan J, Lu T, Lei Q, Chen C, Zeng L - Oxid Med Cell Longev (2015)

Effects of Hsp20 on GA morphology and apoptosis in N2a cells exposed to OGDR. N2a cells were transfected with different plasmids (pEGFP-N1 and pEGFP-Hsp20). After transfection for 36 h, cells were treated with OGD 4 h plus 12 h reperfusion. The experiment was repeated independently for at least three times. (a) Digital photomicrograph under fluorescent illumination showing the morphology of GA was detected using Golgi58 staining. GA displayed typical ribbon-like structures adjacent to the nuclei in N2a cell transfected with pEGFP-N1 without OGDR exposure. Fragmented GA was evident in N2a cells transfected with pEGFP-N1 exposed to 4 h OGD plus 12 h reperfusion insult. Transfection with Hsp20 significantly attenuated OGDR-induced fragmentation of GA. (b) Quantitation (mean ± SEM) of (a) from three independent experiments. (c) Overexpression of Hsp20 in vitro reduced OGDR-induced apoptosis. Values are expressed as mean ± SEM. ∗P < 0.05 compared to control. ∗∗P < 0.01 compared to control (Bar = 10 μm).
© Copyright Policy - open-access
Related In: Results  -  Collection

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fig3: Effects of Hsp20 on GA morphology and apoptosis in N2a cells exposed to OGDR. N2a cells were transfected with different plasmids (pEGFP-N1 and pEGFP-Hsp20). After transfection for 36 h, cells were treated with OGD 4 h plus 12 h reperfusion. The experiment was repeated independently for at least three times. (a) Digital photomicrograph under fluorescent illumination showing the morphology of GA was detected using Golgi58 staining. GA displayed typical ribbon-like structures adjacent to the nuclei in N2a cell transfected with pEGFP-N1 without OGDR exposure. Fragmented GA was evident in N2a cells transfected with pEGFP-N1 exposed to 4 h OGD plus 12 h reperfusion insult. Transfection with Hsp20 significantly attenuated OGDR-induced fragmentation of GA. (b) Quantitation (mean ± SEM) of (a) from three independent experiments. (c) Overexpression of Hsp20 in vitro reduced OGDR-induced apoptosis. Values are expressed as mean ± SEM. ∗P < 0.05 compared to control. ∗∗P < 0.01 compared to control (Bar = 10 μm).
Mentions: To further analyze the effects of Hsp20 on OGDR-induced Golgi fragmentation, N2a cells were transfected with different plasmids (pEGFP-N1 and pEGFP-Hsp20). After transfection for 36 h, cells were treated with OGD 4 h plus 12 h reperfusion. Results showed that, in N2a cell transfected with empty vector (pEGFP-N1) without OGDR exposure, GA displayed typical ribbon-like structures adjacent to the nuclei (Figure 3(a)). When N2a cell transfected with pEGFP-N1 were exposed to OGD 4 h plus 12 h reperfusion, lots of GA changed to debris-like structures scattered in the cytoplasm. Meanwhile, transfection with Hsp20 significantly attenuated OGDR-induced Golgi fragmentation (Figures 3(a) and 3(b)).

Bottom Line: Finding effective neuroprotective agents has become a priority in the treatment of ischemic stroke.However, transfection with Hsp20 significantly attenuates OGDR-induced Golgi fragmentation and apoptosis.Hsp20 interacts with Bax, decreases FasL and Bax expression, and inhibits caspases 3 and p115 cleavage in N2a cells exposed to OGDR.

View Article: PubMed Central - PubMed

Affiliation: Department of Neurology, Second Xiangya Hospital, Central South University, Changsha, Hunan 410011, China ; Department of Traditional Chinese Medicine, Second Xiangya Hospital, Central South University, Changsha, Hunan 410011, China.

ABSTRACT
Cerebral ischemia-reperfusion injury plays an important role in the development of tissue injury after acute ischemic stroke. Finding effective neuroprotective agents has become a priority in the treatment of ischemic stroke. The Golgi apparatus (GA) is a pivotal organelle and its protection is an attractive target in the treatment of cerebral ischemia-reperfusion injury. Protective effects of Hsp20, a potential cytoprotective agent due to its chaperone-like activity and involvement in regulation of many vital processes, on GA were assessed in an ischemia-reperfusion injury model. Mouse neuroblastoma Neuro2a (N2a) cells were subjected to oxygen-glucose deprivation/reperfusion (OGDR) insult. OGDR induces Golgi fragmentation, apoptosis, and p115 cleavage in N2a cells. However, transfection with Hsp20 significantly attenuates OGDR-induced Golgi fragmentation and apoptosis. Hsp20 interacts with Bax, decreases FasL and Bax expression, and inhibits caspases 3 and p115 cleavage in N2a cells exposed to OGDR. Our data demonstrate that increased Hsp20 expression protects against OGDR-induced Golgi fragmentation and apoptosis, likely through interaction with Bax and subsequent amelioration of the OGDR-induced elevation in p115 cleavage via the Fas/FasL signaling pathway. This neuroprotective potential of Hsp20 against OGDR insult and the underlying mechanism will pave the way for its potential clinical application for cerebral ischemia-reperfusion related disorders.

No MeSH data available.


Related in: MedlinePlus