Background： Bovine milk contains not only a variety of nutritional ingredients but also microRNAs （miRNAs） that are thought to be secreted by the bovine mammary epithelial cells （BMECs）. The objective of this stu...Background： Bovine milk contains not only a variety of nutritional ingredients but also microRNAs （miRNAs） that are thought to be secreted by the bovine mammary epithelial cells （BMECs）. The objective of this study was to elucidate the production of milk-related miRNAs in BMECs under the influence of lactogenic hormones. Results： According to a microarray result of milk exosomal miRNAs prior to cellular analyses, a total of 257 miRNAs were detected in a Holstein cow milk. Of these, 18 major miRNAs of interest in the milk were selected for an expression analysis in BMEC culture that was treated with or without dexamethasone, insulin, and prolactin （DIP） to induce a lactogenic differentiation. Quantitative polymerase chain reaction （qPCR） results showed that the expressions of miR-21-Sp （P = 0.005）, miR-26a （P = 0.016）, and miR-320a （P = 0.011） were lower in the DIP-treated cells than in the untreated cells. In contrast, the expression of miR-339a （P-- 0.017） in the cell culture medium were lower in the DiP-treated culture than in the untreated culture. Intriguingly, the miR-148a expression in cell culture medium was elevated by DIP treatment of BMEC culture （P = 0.018）. The medium-to-cell expression ratios of miR- 103 （P = 0.025）, miR-148a （P 〈 0.001）, and miR-223 （P = 0.013） were elevated in the DIP-treated BMECs, suggesting that the lactogenic differentiation-induced secretion of these three miRNAs in BMECs. A bioinformatic analysis showed that the miRNAs down-regulated in the BMECs were associated with the suppression of genes related to transcriptional regulation, protein phosphorylation, and tube development. Conclusion： The results suggest that the miRNAs changed by lactogenic hormones are associated with milk protein synthesis, and mammary gland development and maturation. The elevated miR-148a level in DIP-treated BMECs may be associated with its increase in milk during the lactation period of cows.展开更多
Porcine epidemic diarrhea(PED)caused by porcine epidemic diarrhea virus(PEDV)is one of the most devastating diseases in the global pig industry due to its high mortality rate in piglets.Maternal vaccines can effective...Porcine epidemic diarrhea(PED)caused by porcine epidemic diarrhea virus(PEDV)is one of the most devastating diseases in the global pig industry due to its high mortality rate in piglets.Maternal vaccines can effectively enhance the gut-mammary gland-secretory IgA axis to boost lactogenic immunity and passive protection of nursing piglets against PEDV challenge.From 2017 to 2021,we collected 882 diarrhea samples from 303 farms in China to investigate the epidemiology of PEDV.The result showed that about 52.15%(158/303)of the farms were positive for PEDV with an overall detection rate of 63.95%(564/882)of the samples.The S1 fragments of S gene from 104 strains were sequenced for the phylogenetic analysis.A total of 71 PEDV strains(68.27%)sequenced in this study were clustered into the predominant G2c subgroup,while the newly-defined G2d strains(9.62%)were identified in three provinces of China.The NH-TA2020 strain of G2c subgroup was isolated and cultured,and its infection to piglets caused watery diarrhea within 24 h,indicating its strong pathogenicity.Oral administration of NH-TA2020 strain to pregnant gilts stimulated high levels of IgA antibody in colostrum.The piglets fed by the gilts above were challenged with NH-TA2020 strain or CH-HeB-RY-2020 strain from G2d subgroup,and the clinical symptoms and virus shedding were significantly reduced compared to the mock group.Our findings suggest that G2c subgroup is the predominant branch circulating in China from 2017 to 2021.Oral administration of NH-TA2020 enhances maternal IgA and lactogenic immune responses,which confer protection against the homologous and emerging G2d PEDV strains challenges in neonates.展开更多
基金supported by the Japan Society for the Promotion of Science (JSPS KAKENHI 25660220) to S.M.
文摘Background： Bovine milk contains not only a variety of nutritional ingredients but also microRNAs （miRNAs） that are thought to be secreted by the bovine mammary epithelial cells （BMECs）. The objective of this study was to elucidate the production of milk-related miRNAs in BMECs under the influence of lactogenic hormones. Results： According to a microarray result of milk exosomal miRNAs prior to cellular analyses, a total of 257 miRNAs were detected in a Holstein cow milk. Of these, 18 major miRNAs of interest in the milk were selected for an expression analysis in BMEC culture that was treated with or without dexamethasone, insulin, and prolactin （DIP） to induce a lactogenic differentiation. Quantitative polymerase chain reaction （qPCR） results showed that the expressions of miR-21-Sp （P = 0.005）, miR-26a （P = 0.016）, and miR-320a （P = 0.011） were lower in the DIP-treated cells than in the untreated cells. In contrast, the expression of miR-339a （P-- 0.017） in the cell culture medium were lower in the DiP-treated culture than in the untreated culture. Intriguingly, the miR-148a expression in cell culture medium was elevated by DIP treatment of BMEC culture （P = 0.018）. The medium-to-cell expression ratios of miR- 103 （P = 0.025）, miR-148a （P 〈 0.001）, and miR-223 （P = 0.013） were elevated in the DIP-treated BMECs, suggesting that the lactogenic differentiation-induced secretion of these three miRNAs in BMECs. A bioinformatic analysis showed that the miRNAs down-regulated in the BMECs were associated with the suppression of genes related to transcriptional regulation, protein phosphorylation, and tube development. Conclusion： The results suggest that the miRNAs changed by lactogenic hormones are associated with milk protein synthesis, and mammary gland development and maturation. The elevated miR-148a level in DIP-treated BMECs may be associated with its increase in milk during the lactation period of cows.
基金supported by the National Key R&D Program of China(2021ZD0113803)the"Pioneer"and"Leading Goose"R&D Program of Zhejiang (2022C02031)the National Natural Science Foundation of China (No. 31701424)
文摘Porcine epidemic diarrhea(PED)caused by porcine epidemic diarrhea virus(PEDV)is one of the most devastating diseases in the global pig industry due to its high mortality rate in piglets.Maternal vaccines can effectively enhance the gut-mammary gland-secretory IgA axis to boost lactogenic immunity and passive protection of nursing piglets against PEDV challenge.From 2017 to 2021,we collected 882 diarrhea samples from 303 farms in China to investigate the epidemiology of PEDV.The result showed that about 52.15%(158/303)of the farms were positive for PEDV with an overall detection rate of 63.95%(564/882)of the samples.The S1 fragments of S gene from 104 strains were sequenced for the phylogenetic analysis.A total of 71 PEDV strains(68.27%)sequenced in this study were clustered into the predominant G2c subgroup,while the newly-defined G2d strains(9.62%)were identified in three provinces of China.The NH-TA2020 strain of G2c subgroup was isolated and cultured,and its infection to piglets caused watery diarrhea within 24 h,indicating its strong pathogenicity.Oral administration of NH-TA2020 strain to pregnant gilts stimulated high levels of IgA antibody in colostrum.The piglets fed by the gilts above were challenged with NH-TA2020 strain or CH-HeB-RY-2020 strain from G2d subgroup,and the clinical symptoms and virus shedding were significantly reduced compared to the mock group.Our findings suggest that G2c subgroup is the predominant branch circulating in China from 2017 to 2021.Oral administration of NH-TA2020 enhances maternal IgA and lactogenic immune responses,which confer protection against the homologous and emerging G2d PEDV strains challenges in neonates.
