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Localization and expression of Hsp27 and αB-crystallin in rat primary myocardial cells during heat stress in vitro.

Tang S, Buriro R, Liu Z, Zhang M, Ali I, Adam A, Hartung J, Bao E - PLoS ONE (2013)

Bottom Line: After exposure to heat stress at 42°C for different durations, the activities of enzymes expressed during cell damage increased in the supernatant of the heat-stressed myocardial cells from 10 min, and the pathological lesions were characterized by karyopyknosis and acute degeneration.According to the Western blotting results, during the 480 min of heat stress, no significant variation was found in Hsp27 and αB-crystallin expression; however, significant differences were found in the induction of their corresponding mRNAs.Our findings indicate that Hsp27 and αB-crystallin do play a role in the response of cardiac cells to heat stress, but the details of their function remain to be investigated.

View Article: PubMed Central - PubMed

Affiliation: College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, China.

ABSTRACT
Neonatal rat primary myocardial cells were subjected to heat stress in vitro, as a model for investigating the distribution and expression of Hsp27 and αB-crystallin. After exposure to heat stress at 42°C for different durations, the activities of enzymes expressed during cell damage increased in the supernatant of the heat-stressed myocardial cells from 10 min, and the pathological lesions were characterized by karyopyknosis and acute degeneration. Thus, cell damage was induced at the onset of heat stress. Immunofluorescence analysis showed stronger positive signals for both Hsp27 and αB-crystallin from 10 min to 240 min of exposure compared to the control cells. According to the Western blotting results, during the 480 min of heat stress, no significant variation was found in Hsp27 and αB-crystallin expression; however, significant differences were found in the induction of their corresponding mRNAs. The expression of these small heat shock proteins (sHsps) was probably delayed or overtaxed due to the rapid consumption of sHsps in myocardial cells at the onset of heat stress. Our findings indicate that Hsp27 and αB-crystallin do play a role in the response of cardiac cells to heat stress, but the details of their function remain to be investigated.

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Transcription levels of hsp27 mRNA and αB-crystallin mRNA in myocardial cells after heat stress.qPCR Ct values for hsp27 and αB-crystallin were normalized against β-actin. Expression of this housekeeping gene did not change in response to heat stress. The results showed that the transcription levels of hsp27 mRNA increased immediately and significantly (P<0.01) from 10 min after the onset of heat shock until 480 min, peaking at 360 min. The transcription levels of αB-crystallin mRNA also showed a significant (P<0.01) increase from 10 min after exposure to heat stress until 480 min, and peaked at 360 min. For αB-crystallin mRNA, aP<0.05; a*, P<0.01. For hsp27 mRNA, bP<0.01.
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pone-0069066-g005: Transcription levels of hsp27 mRNA and αB-crystallin mRNA in myocardial cells after heat stress.qPCR Ct values for hsp27 and αB-crystallin were normalized against β-actin. Expression of this housekeeping gene did not change in response to heat stress. The results showed that the transcription levels of hsp27 mRNA increased immediately and significantly (P<0.01) from 10 min after the onset of heat shock until 480 min, peaking at 360 min. The transcription levels of αB-crystallin mRNA also showed a significant (P<0.01) increase from 10 min after exposure to heat stress until 480 min, and peaked at 360 min. For αB-crystallin mRNA, aP<0.05; a*, P<0.01. For hsp27 mRNA, bP<0.01.

Mentions: The transcription levels of hsp27 mRNA and αB-crystallin mRNA, normalized to myocardial cell β-actin mRNA, are displayed in Figure 5. Ct values for hsp27 and αB-crystallin were normalized against β-actin. The expression of this housekeeping gene did not change in response to heat stress. The transcription levels of hsp27 mRNA were increased immediately and significantly (P<0.01) after exposure to heat shock. With increase in the amount of exposure time, the transcription levels of hsp27 mRNA persistently increased. The transcription level of hsp27 mRNA was eight-fold higher at 120 min than that at 60 min of heat stress, and reached peak levels at 360 min, at which point it was 18-fold higher than that at 240 min. The transcription levels of αB-crystallin mRNA in the myocardial cells showed a significant (P<0.01) increase as soon as they were exposed to high temperature (at 10 min). After 120 min of heat exposure, the transcription level of αB-crystallin mRNA in the heat-stressed myocardial cells was four-fold higher, and it reached the highest level at 360 min of exposure.


Localization and expression of Hsp27 and αB-crystallin in rat primary myocardial cells during heat stress in vitro.

Tang S, Buriro R, Liu Z, Zhang M, Ali I, Adam A, Hartung J, Bao E - PLoS ONE (2013)

Transcription levels of hsp27 mRNA and αB-crystallin mRNA in myocardial cells after heat stress.qPCR Ct values for hsp27 and αB-crystallin were normalized against β-actin. Expression of this housekeeping gene did not change in response to heat stress. The results showed that the transcription levels of hsp27 mRNA increased immediately and significantly (P<0.01) from 10 min after the onset of heat shock until 480 min, peaking at 360 min. The transcription levels of αB-crystallin mRNA also showed a significant (P<0.01) increase from 10 min after exposure to heat stress until 480 min, and peaked at 360 min. For αB-crystallin mRNA, aP<0.05; a*, P<0.01. For hsp27 mRNA, bP<0.01.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC3716771&req=5

pone-0069066-g005: Transcription levels of hsp27 mRNA and αB-crystallin mRNA in myocardial cells after heat stress.qPCR Ct values for hsp27 and αB-crystallin were normalized against β-actin. Expression of this housekeeping gene did not change in response to heat stress. The results showed that the transcription levels of hsp27 mRNA increased immediately and significantly (P<0.01) from 10 min after the onset of heat shock until 480 min, peaking at 360 min. The transcription levels of αB-crystallin mRNA also showed a significant (P<0.01) increase from 10 min after exposure to heat stress until 480 min, and peaked at 360 min. For αB-crystallin mRNA, aP<0.05; a*, P<0.01. For hsp27 mRNA, bP<0.01.
Mentions: The transcription levels of hsp27 mRNA and αB-crystallin mRNA, normalized to myocardial cell β-actin mRNA, are displayed in Figure 5. Ct values for hsp27 and αB-crystallin were normalized against β-actin. The expression of this housekeeping gene did not change in response to heat stress. The transcription levels of hsp27 mRNA were increased immediately and significantly (P<0.01) after exposure to heat shock. With increase in the amount of exposure time, the transcription levels of hsp27 mRNA persistently increased. The transcription level of hsp27 mRNA was eight-fold higher at 120 min than that at 60 min of heat stress, and reached peak levels at 360 min, at which point it was 18-fold higher than that at 240 min. The transcription levels of αB-crystallin mRNA in the myocardial cells showed a significant (P<0.01) increase as soon as they were exposed to high temperature (at 10 min). After 120 min of heat exposure, the transcription level of αB-crystallin mRNA in the heat-stressed myocardial cells was four-fold higher, and it reached the highest level at 360 min of exposure.

Bottom Line: After exposure to heat stress at 42°C for different durations, the activities of enzymes expressed during cell damage increased in the supernatant of the heat-stressed myocardial cells from 10 min, and the pathological lesions were characterized by karyopyknosis and acute degeneration.According to the Western blotting results, during the 480 min of heat stress, no significant variation was found in Hsp27 and αB-crystallin expression; however, significant differences were found in the induction of their corresponding mRNAs.Our findings indicate that Hsp27 and αB-crystallin do play a role in the response of cardiac cells to heat stress, but the details of their function remain to be investigated.

View Article: PubMed Central - PubMed

Affiliation: College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, China.

ABSTRACT
Neonatal rat primary myocardial cells were subjected to heat stress in vitro, as a model for investigating the distribution and expression of Hsp27 and αB-crystallin. After exposure to heat stress at 42°C for different durations, the activities of enzymes expressed during cell damage increased in the supernatant of the heat-stressed myocardial cells from 10 min, and the pathological lesions were characterized by karyopyknosis and acute degeneration. Thus, cell damage was induced at the onset of heat stress. Immunofluorescence analysis showed stronger positive signals for both Hsp27 and αB-crystallin from 10 min to 240 min of exposure compared to the control cells. According to the Western blotting results, during the 480 min of heat stress, no significant variation was found in Hsp27 and αB-crystallin expression; however, significant differences were found in the induction of their corresponding mRNAs. The expression of these small heat shock proteins (sHsps) was probably delayed or overtaxed due to the rapid consumption of sHsps in myocardial cells at the onset of heat stress. Our findings indicate that Hsp27 and αB-crystallin do play a role in the response of cardiac cells to heat stress, but the details of their function remain to be investigated.

Show MeSH
Related in: MedlinePlus