Wnt5a介导Wnt/Ca2+通路对BCG诱导牛原代肺泡上皮细胞自噬的调控作用

doi:10.11843/j.issn.0366-6964.2022.11.032

Abstract

Abstract: The aim of this study was to investigate the method of in vitro isolation and culture of bovine alveolar epithelial cells (BAECs) and the regulatory mechanism of Wnt5a on the autophagy of BAECs infected with Mycobacterium tuberculosis bacille Calmette-Guérin (BCG). BAECs were isolated by enzyme combined digestion method and mechanical scraping method, and purified by differential adhesion method. The expression of epithelial cell markers CK14 and CK5 were detected by immunofluorescence staining. BAECs were infected with BCG and the expression of Wnt5a was inhibited by Box-5. The expression of autophagy-related proteins and nonclassical Wnt signaling pathway related proteins were detected by Western blot and immunofluorescence staining. The results showed that BAECs were successfully isolated with high purity by enzyme combined digestion and mechanical scraping. Isolated cells were identified as positive by CK14 and CK5. BCG infection with BAECs promoted the expression of Wnt5a and increased autophagy. Box-5 pretreatment down-regulated the expression of BCG-induced autophagy-related proteins LC3II, P62, Atg7 and Atg5, and inhibited the expression of nonclassical Wnt/Ca²⁺ signaling pathway related proteins Wnt5a, CaMKII and NFAT. In conclusion, the isolation and culture method of BAECs was successfully established. BCG infection promoted the Wnt5a expression and autophagy of BAECs, and the inhibition of Wnt5a expression down-regulated the BCG-induced autophagy in BAECs. Moreover, Wnt5a regulates the autophagy induced by BCG in BAECs through the nonclassical Wnt/Ca²⁺ signaling pathway.

Key words: Wnt5a, BCG, bovine alveolar epithelial cells, autophagy

CLC Number:

S852.4

CHEN Qi, DU Changzheng, ZHENG Xuedi, MA Boli, XU Jinrui, YANG Yi. Regulation on BCG-induced Autophagy by Wnt5a-mediated Wnt/Ca²⁺ Pathway in BAECs[J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(11): 4071-4080.

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Regulation on BCG-induced Autophagy by Wnt5a-mediated Wnt/Ca²⁺ Pathway in BAECs

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Regulation on BCG-induced Autophagy by Wnt5a-mediated Wnt/Ca2+ Pathway in BAECs

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Regulation on BCG-induced Autophagy by Wnt5a-mediated Wnt/Ca²⁺ Pathway in BAECs