Selenium(Se)deficiency can seriously affect the small intestine of swine,and cause diarrhea in swine.However,the specific mechanism of Se deficiency-induced swine diarrhea has rarely been reported.Here,to explore the ...Selenium(Se)deficiency can seriously affect the small intestine of swine,and cause diarrhea in swine.However,the specific mechanism of Se deficiency-induced swine diarrhea has rarely been reported.Here,to explore the damage of Se deficiency on the calcium homeostasis and autophagy mechanism of swine,in vivo and in vitro models of swine intestinal Se deficiency were established.Twenty-four pure line castrated male Yorkshire pigs(45 d old,12.50±1.32 kg,12 full-sibling pairs)were divided into 2 equal groups and fed Se-deficient diet(0.007 mg Se/kg)as the Se-deficiency group,or fed Se-adequate diet(0.3 mg Se/kg)as the control group for 16 weeks.The intestinal porcine enterocyte cell line(IPEC-J2)was divided into 2 groups,and cultured by Se-deficient medium as the Se-deficient group,or cultured by normal medium as the control group.Morphological observations showed that compared with the control group,intestinal cells in the Se-deficiency group were significantly damaged,and autophagosomes increased.Autophagy staining and cytoplasmic calcium staining results showed that in the Sedeficiency group,autophagy increased and calcium homeostasis was destroyed.According to the reactive oxygen species(ROS)staining results,the percentage of ROS in the Se-deficiency group was higher than that in the control group in the in vitro model.Compared with the control group,the protein and mRNA expressions of autophagy-calcium-related genes including Beclin 1,microtubule-associated proteins 1 A(LC3-1),microtubule-associated proteins 1 B(LC3-2),autophagy-related protein 5(ATG5),autophagy-related protein 12(ATG12),autophagy-related protein 16(ATG16),mammalian target of rapamycin(mTOR),calmodulin-dependent protein kinase kinaseβ(CAMKK-β),adenosine 5’-monophosphate-activated protein kinase(AMPK),sarco(endo)plasmic reticulum Ca2+-ATPase(SERCA),and calpain in the Se-deficiency group were significantly increased which was consistent in vivo and in vitro(P<0.05).Altogether,our results indicated that Se deficiency could destroy the calcium homeostasis of the swine small intestine to trigger cell autophagy and oxidative stress,which was helpful to explain the mechanism of Se deficiency-induced diarrhea in swine.展开更多
基金supported by the Natural Science Foundation of Heilongjiang Province of China(YQ2021C021)。
文摘Selenium(Se)deficiency can seriously affect the small intestine of swine,and cause diarrhea in swine.However,the specific mechanism of Se deficiency-induced swine diarrhea has rarely been reported.Here,to explore the damage of Se deficiency on the calcium homeostasis and autophagy mechanism of swine,in vivo and in vitro models of swine intestinal Se deficiency were established.Twenty-four pure line castrated male Yorkshire pigs(45 d old,12.50±1.32 kg,12 full-sibling pairs)were divided into 2 equal groups and fed Se-deficient diet(0.007 mg Se/kg)as the Se-deficiency group,or fed Se-adequate diet(0.3 mg Se/kg)as the control group for 16 weeks.The intestinal porcine enterocyte cell line(IPEC-J2)was divided into 2 groups,and cultured by Se-deficient medium as the Se-deficient group,or cultured by normal medium as the control group.Morphological observations showed that compared with the control group,intestinal cells in the Se-deficiency group were significantly damaged,and autophagosomes increased.Autophagy staining and cytoplasmic calcium staining results showed that in the Sedeficiency group,autophagy increased and calcium homeostasis was destroyed.According to the reactive oxygen species(ROS)staining results,the percentage of ROS in the Se-deficiency group was higher than that in the control group in the in vitro model.Compared with the control group,the protein and mRNA expressions of autophagy-calcium-related genes including Beclin 1,microtubule-associated proteins 1 A(LC3-1),microtubule-associated proteins 1 B(LC3-2),autophagy-related protein 5(ATG5),autophagy-related protein 12(ATG12),autophagy-related protein 16(ATG16),mammalian target of rapamycin(mTOR),calmodulin-dependent protein kinase kinaseβ(CAMKK-β),adenosine 5’-monophosphate-activated protein kinase(AMPK),sarco(endo)plasmic reticulum Ca2+-ATPase(SERCA),and calpain in the Se-deficiency group were significantly increased which was consistent in vivo and in vitro(P<0.05).Altogether,our results indicated that Se deficiency could destroy the calcium homeostasis of the swine small intestine to trigger cell autophagy and oxidative stress,which was helpful to explain the mechanism of Se deficiency-induced diarrhea in swine.
