Transcription pattern of key molecular chaperones in heat shocked caprine cardiac fibroblasts.

The present study was attempted to unveil the impact of heat stress on transcription pattern of major heat shock response genes in caprine cardiac fibroblasts. Cardiac tissues (n = 6) were collected and primary cardiac cell culture was done. Cultured cardiac fibroblasts were kept in an atmosphere of 5% CO2 and 95% air at 38.5 °C. Cardiac cells achieved 70-75% confluence after 72 hours of incubation. Heat stress was induced on confluent cardiac fibroblasts at 42 °C for 0 (control), 20, 60, 100 and 200 min. Quantitative RT-PCR for ß2m (internal control), HSP60, HSP70, HSP90, and HSP110 was done and their transcription pattern was assessed by Pfaffl method. HSP60, HSP90, and HSP110 transcription did not differ at 20 min, up-regulated (p < 0.05) from 60 to 200 min and registered highest at 200 min of heat exposure. HSP70 transcription was gradually escalated (p < 0.05) time dependently from 20 to 200 min and reached zenith at 200 min of heat exposure. Differential induction in transcription of key molecular chaperones at various durations of heat exposure might reduce cardiac fibroblasts apoptosis and thus could maintain cardiac tissue function during heat stress.

Hyper-transcription of heat shock factors and heat shock proteins safeguard caprine cardiac cells against heat stress.

The present study was undertaken to document the transcriptional abundance of heat shock factors and heat shock proteins and their role in survivability of caprine cardiac cells during heat stress. Cardiac tissues were collected from different goats (n = 6) and primary cardiac cell culture was done in an atmosphere of 5% CO2 and 95% air at 38.5 °C. Cardiac cells accomplished 70-75% confluence after 72 h of incubation. Confluent cardiac cells were exposed to heat stress at 42 °C for 0 (control), 20, 60, 100 and 200 min. Quantitative RT-PCR for ß2m (internal control), heat shock factors (HSF1, HSF2, HSF4, HSF5), heat shock proteins (HSP10, HSP40), and Caspase-3 was done and their transcriptional abundance was assessed by Pfaffl method. Transcriptional abundance of HSF1, HSF2, and HSF4 did not change at 20 min, increased (P < 0.05) from 60 to 200 min and reached zenith at 200 min of heat exposure. However, transcriptional abundance of HSF5 was gradually escalated (P < 0.05) from 20 to 200 min and registered highest at 200 min of heat exposure. Transcriptional abundance of HSP10 and HSP40 followed an similar pattern like that of HSF5. Transcriptional abundance of Caspase-3 was significantly down-regulated at 200 min of heat exposure. It could be speculated that over-expression of HSFs and HSPs might have reduced Caspase-3 expression at 200 min of heat exposure suggesting their involvement in cardiac cells survival under heat stress. Moreover, hyper-expression of HSFs and HSPs could maintain the integrity and endurance of cardiac tissues of goats under heat stress.