Satapathy P P, Mishra S R, Jena G R, Kundu A K
Department of Veterinary Physiology, C.V.Sc & A.H., O.U.A.T, Bhubaneswar, 751003, India.
Department of Veterinary Physiology, C.V.Sc & A.H., O.U.A.T, Bhubaneswar, 751003, India.
J Therm Biol. 2023 Jan;111:103393. doi: 10.1016/j.jtherbio.2022.103393. Epub 2022 Nov 30.
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% CO 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 β2m (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.
本研究旨在记录热休克因子和热休克蛋白的转录丰度及其在热应激期间山羊心脏细胞存活能力中的作用。从不同的山羊(n = 6)收集心脏组织,并在38.5°C、5% CO和95%空气的环境中进行原代心脏细胞培养。培养72小时后,心脏细胞达到70 - 75%汇合度。将汇合的心脏细胞在42°C下暴露于热应激0(对照)、20、60、100和200分钟。对β2m(内参)、热休克因子(HSF1、HSF2、HSF4、HSF5)、热休克蛋白(HSP10、HSP40)和Caspase-3进行定量RT-PCR,并通过Pfaffl方法评估它们的转录丰度。HSF1、HSF2和HSF4的转录丰度在20分钟时没有变化,从60分钟到200分钟增加(P < 0.05),并在热暴露200分钟时达到峰值。然而,HSF5的转录丰度从20分钟到200分钟逐渐升高(P < 0.05),并在热暴露200分钟时达到最高。HSP10和HSP40的转录丰度遵循与HSF5相似的模式。Caspase-3的转录丰度在热暴露200分钟时显著下调。可以推测,热暴露200分钟时热休克因子和热休克蛋白的过表达可能降低了Caspase-3的表达,表明它们参与了热应激下心脏细胞的存活。此外,热休克因子和热休克蛋白的过度表达可以维持热应激下山羊心脏组织的完整性和耐受性。