The Effect of Zearalenone on the Growth of Bovine Mammary Epithelial Cells and the Expression of Genes Related to Milk Fat Synthesis

doi:10.7668/hbnxb.20194501

Abstract

Abstract:

This study aims to explore the effects of different concentrations of zearalenone (ZEN) on the growth of bovine mammary alveolar cell-T (MAC-T) and the expression of genes related to milk fat synthesis.Firstly,MAC-T cells were treated with different doses of ZEN for 36 hours,and the number of cells was counted using a hemocytometer.Then,cells were stained and analyzed for apoptosis and necrosis to determine the appropriate dose of ZEN.Next,acommercial detection kit was used to examine the impact of ZEN on reactive oxygen species (ROS) and mitochondria number in MAC-T cells.Finally,using Real-time Quantitative PCR (RT-qPCR) technology,the influence of ZEN on proliferation,apoptosis,oxidative stress,and milk fat synthesis-related gene expression in MAC-T cells was analyzed.The results showed that low doses of ZEN (0.01—1.00 μmol/L) tended to promote MAC-T growth,whereas high doses of ZEN (5.00—10.00 μmol/L) significantly reduced the number of cells,0.10 μmol/L ZEN had no obvious effect on ROS and mitochondria number in MAC-T,but 10.00 μmol/L ZEN notably elevated ROS levels and decreased mitochondria number.RT-qPCR results indicated that 0.10 μmol/L ZEN significantly promoted the expression of proliferation genes (CDK1,CCND2),antioxidant genes (DHODH,GPX4),and anti-apoptotic genes (BCL-2),but also increased the expression of apoptotic genes (CAS-3,BAX).Whereas,10.00 μmol/L ZEN significantly inhibited the expression of proliferation genes (PCNA,CDK1,CCND2),antioxidant genes (DHODH,GPX4,AIFM2),and anti-apoptotic genes (BCL-2),while significantly promoting the expression of apoptotic genes (CAS-3,BAX).Notably,0.10 μmol/L ZEN obviously promoted the expression of milk fat synthesis-related genes (PPARγ,FASN,JAK-2),but 10.00 μmol/L ZEN significantly suppressed these genes.The aforementioned results suggested different effects of ZEN concentrations on MAC-T cells:0.10 μmol/L ZEN could promote MAC-T cell growth and milk fat synthesis-related gene expression while also inducing the expression of apoptotic genes.In contrast,10.00 μmol/L ZEN induces oxidative stress in MAC-T cells,reduces mitochondria number,inhibits the expression of genes associated with proliferation,antioxidant properties,anti-apoptotic properties,and milk fat synthesis,while promoting the expression of apoptotic genes,leading to cell apoptosis.

Key words: Milk, Zearalenone, Bovine mammary epithelial cells, Milk fat synthesis, Proliferation, Apoptosis

MA Zifeng, LI Qiao, XU Hongmei, LI Yueyue, YIN Shi, HE Honghong, XIONG Yan, LAN Daoliang, LI Jian, XIONG Xianrong, FU Wei. The Effect of Zearalenone on the Growth of Bovine Mammary Epithelial Cells and the Expression of Genes Related to Milk Fat Synthesis[J]. Acta Agriculturae Boreali-Sinica, 2024, 39(1): 201-210. doi: 10.7668/hbnxb.20194501.

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References 23

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基因 Gene	引物序列(5'—3') Primer sequences(5'—3')	NCBI 序列号 NCBI sequence ID	产物长度/bp Product size	退火温度/℃ Tm
GAPDH	F:ACAGTCAAGGCAGAGAACGG R:GGTTCACGCCCATCACAAAC	XM_027541111.1	235	60
GPX4	F:TGTGGTTTACGGATCCTGGC R:CCCTTGGGCTGGACTTTCAT	NM_001346431.1	182	60
JAK-2	F:GAGGCACCGGTTTCAGAAG R:AGCCATTCCCATGCAGAGTC	XM_024986698.1	208	60
DHODH	F:GGAGCTGCGTCCTTGATTCT R:TTCTAGGCAAGAGGGTTGCG	NM_001015650.1	141	60
PPARγ	F:GATAGGTGTGATCTTAACTGTCGGAT R:CGCTAACAGCTTCTCCTTCTCG	NM_181024.2	150	60
FASN	F:CTGGAGCGTGAGCACAACCTG R:GTGTGGAGTCCGTCAGCTCAT	XM_005220997.3	118	60
BAX	F:GTGCCCGAGTTGATCAGGAC R:CCATGTGGGTGTCCCAAAGT	XM_015458140.2	175	60
BCL-2	F:GAGTTCGGAGGGGTCATGTG R:GCCTTCAGAGACAGCCAGGA	NM_001166486.1	203	60
PCNA	F:CCTTGGTGCAGCTAACCCTT R:TTGGACATGCTGGTGAGGTT	NM_001034494.1	94	60
CCND2	F:CACCGATGTGGATTGCCTCA R:TCCAGCTCATCCTCCGACTT	NM_001076372.1	117	60
CDK1	F:GCACACAATATGGACCCCCA R:CTGAAGCCTCAGAGCCACTC	NM_001012669.1	139	60
AIFM2	F:TGGCCATGGGGAGAAATGAC R:TTTTCCAGCTCGTGGAGACC	NM_001314031.1	111	60
CAS-3	F:CGAGGCACAGAACTGGACTG R:GCCAGGAAAAGTAACCAGGTG	NM_001077840.1	130	60

基因 Gene	引物序列(5'—3') Primer sequences(5'—3')	NCBI 序列号 NCBI sequence ID	产物长度/bp Product size	退火温度/℃ Tm
GAPDH	F:ACAGTCAAGGCAGAGAACGG R:GGTTCACGCCCATCACAAAC	XM_027541111.1	235	60
GPX4	F:TGTGGTTTACGGATCCTGGC R:CCCTTGGGCTGGACTTTCAT	NM_001346431.1	182	60
JAK-2	F:GAGGCACCGGTTTCAGAAG R:AGCCATTCCCATGCAGAGTC	XM_024986698.1	208	60
DHODH	F:GGAGCTGCGTCCTTGATTCT R:TTCTAGGCAAGAGGGTTGCG	NM_001015650.1	141	60
PPARγ	F:GATAGGTGTGATCTTAACTGTCGGAT R:CGCTAACAGCTTCTCCTTCTCG	NM_181024.2	150	60
FASN	F:CTGGAGCGTGAGCACAACCTG R:GTGTGGAGTCCGTCAGCTCAT	XM_005220997.3	118	60
BAX	F:GTGCCCGAGTTGATCAGGAC R:CCATGTGGGTGTCCCAAAGT	XM_015458140.2	175	60
BCL-2	F:GAGTTCGGAGGGGTCATGTG R:GCCTTCAGAGACAGCCAGGA	NM_001166486.1	203	60
PCNA	F:CCTTGGTGCAGCTAACCCTT R:TTGGACATGCTGGTGAGGTT	NM_001034494.1	94	60
CCND2	F:CACCGATGTGGATTGCCTCA R:TCCAGCTCATCCTCCGACTT	NM_001076372.1	117	60
CDK1	F:GCACACAATATGGACCCCCA R:CTGAAGCCTCAGAGCCACTC	NM_001012669.1	139	60
AIFM2	F:TGGCCATGGGGAGAAATGAC R:TTTTCCAGCTCGTGGAGACC	NM_001314031.1	111	60
CAS-3	F:CGAGGCACAGAACTGGACTG R:GCCAGGAAAAGTAACCAGGTG	NM_001077840.1	130	60

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