热应激对从江香猪十二指肠黏膜结构、HIF-1及其相关蛋白表达的影响

doi:10.11843/j.issn.0366-6964.2024.10.040

Abstract

Abstract:

The aim of this study was to investigate the effects of heat stress (HS) on the duodenal mucosal barrier structure and the character of hypoxia inducible factor 1 (HIF-1) expression and regulation in Congjiang Xiang pigs. Twenty-four fattening pigs were divided into 6 groups randomly, the control group (Con) kept under normal conditions, and the other 5 groups were treated with heat stress for 6, 12, 24, 48, 72 h), respectively. Rectal temperature and respiratory rate were measured during treatment, and the duodenum of each group was collected at the end of the experiment. The change of duodenal tissue structure and goblet cell number in each group were observed with hematoxylin-eosin staining (HE) and alcian blue-periodate Schiff reaction (AB-PAS). The apoptosis cell of duodenal was detected by TUNEL staining. Real-time fluorescence quantification (qRT-PCR), Western blot and immunohistochemistry were used to detect the expression changes of tight junction related proteins, HIF-1 and its upstream regulatory factors respectively. The results showed that, compared with the control group, HS could induce rectal temperature and respiratory rate increase, and the villus height kept decreasing with extension of HS time, while the crypt depth increased synchronously, the ratio of villus height/gland depth and the expression of Occludin and ZO-1 decreased significantly. On the contrary, the number of goblet cells kept increasing. TUNEL showed the apoptosis rate of group HS 72 h was significantly higher than that of control group (P < 0.001).The positive expression of HIF-1α was mainly observed in small intestinal epithelial cells, and which expression enhanced at HS 72 h, the trend was as same as which HIF-1α mRNA and protein expression. The character of HSP90 expression were basically consistent with HIF-1α while the profile of PHD-2 was obviously the opposite. These results indicated that HS could induce apoptosis of small intestinal epithelial cells and damage duodenal mucosal barrier. The increase of HIF-1 expression is closely related to the apoptosis of small intestine cells, and may be regulated by both HSP90 and PHD-2.

Key words: heat stress, Congjiang Xiang pig, intestinal mucosal barrier, hypoxia inducible factor 1(HIF-1), expression regulation

CLC Number:

S828.89

Yongqing LIU, Gang ZHANG, Yanling XIONG, Zhongxin SUN, Fan GAO, Ting LIU, Hui LI. Effects of Heat Stress on Duodenal Mucosal Structure, HIF-1 and Its Related Protein Expression in Congjiang Xiang Pigs[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(10): 4690-4699.

Figures/Tables 7

Table 1

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References 29

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基因名称 Gene name	引物序列(5′→3′) Primer sequence(5′→3′)	产物大小/bp Product size	退火温度/℃ Annealing temperature	登录号 Accesion No.
PHD-2	F：TCATGAGCAGCATGGACGAC	117	63	XM_021073581.1
	R：CGTAACCTGTTCCGTTGCC
HIF-1α	F: TCCTTCGATCAGTTGTCACCATTGG	89	64	NM_001123124.1
	R: GGCATTGGAGTGGGCTGGAATAC
HSP90	F: CAGAGGCGGACAAGAACGACAAG	116	63	NM_213973.2
	R: GATCCTGTTGGCGTGCGTCTG
β-actin	F: TCTGGCACCACACCTTCT	114	60	XM_021086047.1
	R: TGATCTGGGTCATCTTCTCAC

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Effects of Heat Stress on Duodenal Mucosal Structure, HIF-1 and Its Related Protein Expression in Congjiang Xiang Pigs

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