急性热应激对山羊血液生化指标及血淋巴细胞热休克蛋白70家族基因表达的影响

doi:10.11843/j.issn.0366-6964.2019.06.012

摘要/Abstract

摘要： 旨在研究急性热应激对山羊抗氧化能力、免疫功能和血淋巴细胞热休克蛋白70（HSP70）家族基因表达的影响。本试验选取5只健康、体况接近的（12±0.5）月龄波尔山羊×关中奶山羊杂交F₁母羊，饲养于环控舱内（温度维持20℃，相对湿度60%），适应5 d。第6天利用环控舱对5只试验羊进行38℃急性热应激处理12 h，采集热应激前（0 h，20℃）和热应激后2、4、8和12 h试验羊血样。分别利用比色法测定血清抗氧化指标（总抗氧化能力、超氧化物歧化酶、谷胱甘肽过氧化物酶活性和丙二醛含量），ELISA法测定血清免疫指标（免疫球蛋白和细胞因子含量），实时荧光定量PCR法测定血淋巴细胞HSP70家族基因（HSPA1A、HSPA6和HSPA8） mRNA的表达量。结果显示：1）热应激时间对血清抗氧化指标有显著影响。与热应激前0 h相比，血清T-AOC（P<0.05）、SOD（P<0.05）和GSH-Px（P<0.01）活性均在热应激8 h后显著下降，MDA含量在热应激4 h后显著增加（P<0.05）。2）热应激时间对血清免疫指标有显著影响。与热应激前0 h相比，血清TNF-α、IL-1β、IFN-γ和IL-2含量分别在热应激4、8、8和4 h后显著增加（P<0.05）；IL-4（P<0.01）、IgG（P<0.01）、IgM（P<0.01）和IgA（P<0.05）含量分别在热应激12、4、4和4 h后显著下降。3）热应激时间显著提高血淋巴细胞中HSP70家族基因（HSPA1A、HSPA6和HSPA8）的表达量。HSPA1A mRNA表达量呈先升高后下降的趋势，在热应激4 h时达到峰值，各检测时间点均显著高于应激前水平（P<0.01）；HSPA6 mRNA表达量在热应激2 h时显著升高（P<0.01），4 h后恢复到应激前水平（P>0.05）；HSPA8 mRNA表达量在热应激4（P<0.05）、8（P<0.01）、12 h（P<0.01）时显著高于应激前水平。在本试验条件下，38℃急性热应激能够抑制山羊的免疫和抗氧化功能；提高血淋巴细胞HSPA1A、HSPA6和HSPA8基因的表达量，其中HSPA1A对热应激温度和时间更敏感，可作为山羊热应激早期的分子标志物。

Abstract: This experiment was conducted to explore the effects of acute heat stress on antioxidant capacity, immune function and expression of HSP70 family genes in blood lymphocytes in goats. Five (12±0.5)-month-old healthy female goats (F₁ of Boer goat×Guanzhong dairy goat) were selected and raised separately in cage in an environment controlled chamber (temperature:20℃, relative humidity:60%), adaptation period lasted for 5 days. At the 6th day, five goats were treated with acute heat stress at 38℃ for 12 h. Blood samples were collected before heat stress (0 h, 20℃) and after 2, 4, 8 and 12 h of heat stress. Serum antioxidant indexes (total antioxidant capacity, superoxide dismutase, glutathione peroxidase activity and malondialdehyde content), serum immune indexes (serum immunoglobulin and cytokine contents) and the expression of HSP70 family member genes (HSPA1A, HSPA6 and HSPA8) in blood lymphocytes were measured by colorimetric method, ELISA and RT-PCR, respectively. The results showed as follows:1) Heat stress time had significant effects on serum antioxidant indexes. Compared with 0 h before heat stress, serum T-AOC (P<0.05), SOD (P<0.05) and GSH-Px (P<0.01) activities decreased significantly after 8 h of heat stress, and MDA content increased significantly after 4 h of heat stress (P<0.05). 2) Heat stress time had also significant influence on serum immune indexes. The contents of serum TNF-α and IL-2 increased significantly after 4 h of heat stress (P<0.05), while the contents of IL-1β and IFN-γ increased significantly after 8 h of heat stress (P<0.05). The contents of IgG (P<0.01), IgM (P<0.01) and IgA (P<0.05) decreased significantly after 4 h of heat stress, while the content of IL-4 decreased significantly after 12 h of heat stress(P<0.01). 3) Heat stress time significantly increased the expression levels of HSP70 family genes (HSPA1A, HSPA6 and HSPA8) in blood lymphocytes. The mRNA expression of HSPA1A rose firstly, then declined, and then it reached the peak at 4 h after heat stress. In addition, the mRNA expression of HSPA1A was significantly higher at 2, 4, 8 and 12 h after heat stress than at 0 h (P<0.01). The mRNA expression of HSPA6 increased significantly at 2 h after heat stress (P<0.01), and returned to pre-stress level after 4 h (P>0.05). The mRNA expression of HSPA8 was significantly higher at 4 (P<0.05), 8 (P<0.01), 12 h (P<0.01) after heat stress than that at 0 h. In this experiment, acute heat stress at 38℃ could inhibit the immune and antioxidant functions and increased the mRNA expression of HSPA1A, HSPA6 and HSPA8 genes in blood lymphocytes. HSPA1A was more sensitive to temperature and time of heat stress, and could be used as a molecular marker of early heat stress in goats.

中图分类号:

S827.4

彭孝坤, 赵天, 黄晓瑜, 张宇, 邢晓南, 张恩平. 急性热应激对山羊血液生化指标及血淋巴细胞热休克蛋白70家族基因表达的影响[J]. 畜牧兽医学报, 2019, 50(6): 1219-1229.

PENG Xiaokun, ZHAO Tian, HUANG Xiaoyu, ZHANG Yu, XING Xiaonan, ZHANG Enping. Effects of Acute Heat Stress on Blood Biochemistry Indices and Expression of HSP70 Family Genes in Blood Lymphocytes in Goats[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2019, 50(6): 1219-1229.

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