Effects of MitoQ on mitochondrial function and antioxidant capacity of canine bone marrow mesenchymal stem cells

doi:10.3969/j.issn.1004-1524.2022.05.07

Acta Agriculturae Zhejiangensis ›› 2022, Vol. 34 ›› Issue (5): 934-941.DOI: 10.3969/j.issn.1004-1524.2022.05.07

• Animal Science • Previous Articles Next Articles

Effects of MitoQ on mitochondrial function and antioxidant capacity of canine bone marrow mesenchymal stem cells

ZHONG Lijun(), DENG Jiaqiang, GU Congwei, SHEN Liuhong, CAO Suizhong, YU Shumin()

College of Veterinary Medicine,Sichuan Agricultural University,Chengdu 611130,China

Received:2021-01-18 Online:2022-05-25 Published:2022-06-06
Contact: YU Shumin

Abstract

Abstract:

The aim of this study was to investigate the effect of Mitoquinone (MitoQ), a mitochondrial-targeted antioxidant, on the mitochondrial function and antioxidant gene expression of canine bone marrow mesenchymal stem cells (BMSCs), which may provide a theoretical basis for improving the clinical application efficiency of canine BMSCs. Canine BMSCs treated with MitoQ for 48 h were used as materials to evaluate the mitochondrial function and the expression level of antioxidant genes.The results showed that with the increase of passage times,the mitochondrial fusion of BMSCs was significantly advanced, and mitochondrial division was significantly weakened. In addition, the mitochondrial membrane potential and ATP content were significantly decreased and the expression of antioxidant genes of BMSCs was also significantly down-regulated.After MitoQ pretreatment, the expression of BMSCs mitochondrial fusion gene Mfn2 was slightly up-regulated, and the expression of mitochondrial division gene Drp1 and mitochondrial biosynthesis-related gene PGC-1α were significantly up-regulated. More over, the mitochondrial membrane potential and ATP content increased significantly and the expression of antioxidant genes SOD1, SOD2, CAT and GSH-Px was significantly up-regulated.In summary, MitoQ can improve the mitochondrial function and enhance the antioxidant capacity of canine BMSCs.

Key words: canine, mitochondria, oxidative stress

CLC Number:

S829.2

ZHONG Lijun, DENG Jiaqiang, GU Congwei, SHEN Liuhong, CAO Suizhong, YU Shumin. Effects of MitoQ on mitochondrial function and antioxidant capacity of canine bone marrow mesenchymal stem cells[J]. Acta Agriculturae Zhejiangensis, 2022, 34(5): 934-941.

Figures/Tables 6

Table 1 Primer parameters

基因名称Gene ID	上游引物Forward primer (5'→3')	下游引物Reverse primer (5'→3')	长度Length/bp
SOD2	CTCAAGTTCAATGGAGGAGGTCA	CTTGGACACCGACGGATACAG	169
SOD1	CATTACAGGGCTGACTGAAGGC	TTGGACAGAGGATTAAAGTGAGGA	105
CAT	GATACCTGTGAACTGTCCTT	CATTGGCTGCTATGCTCTA	109
GSH-Px	CAACCAGTTCGGGCATCA	CGTTCACCTCGCACTTCTCA	122
Drp1	TAGAAATGCTACTGGTCCTCGTCC	CAGTTCCACACAACGCAGGC	115
Fis1	GTCTGTGGACGACCTGCTGAA	TGCGATGCCTTTACGGATG	142
Mfn2	GGCCTTGATGGGCTACAATGAC	CGCCTCCGACAACGAGAATG	179
Mfn1	CAGGGCGGTTACAGAAGAGGT	CAATACGCTTGGAGTGGGATGA	121
Opa1	GCAGAATCCTAACGCCATCATAC	ACAAATATCGTCCTCCTTCCATG	117
PGC-1α	TTCGGTCATCCCAGTCAAGC	TGTCATCAAACAGGCCATCCA	145
GAPDH	TCCCGCCAACATCAAA	TCACGCCCATCACAAAC	163

Fig.1 Effects of in vitro expansion on morphology of BMSCs A-C represented the morphology of P1, P3 and P6 BMSCs (scale bar=100 μm).

Fig.2 Identification of BMSCs A-F represented the immunofluorescence detection results of cell surface markers CD105, CD90, CD31 and negative control (scale bar=200 μm) and the results of staining after osteogenic differentiation and adipogenic differentiation of BMSCs (scale bar=100 μm).

Fig.3 Effects of in vitro expansion on antioxidant gene expression and mitochondrial function of BMSCs A-F represented the relative expression of SOD1, SOD2, CAT, GSH-Px, PGC-1α, Opa1,Mfn1, Mfn2, Fis1and Drp1,mitochondrial membrane potential level(scale bar=100 μm) and ATP content of BMSCs, respectively. * and **,Significant difference at P<0.05 and P<0.01. ns, No significance. The same as below.

Fig.4 Effects of MitoQ on the expression of BMSCs antioxidant genes and mitochondrial dynamics-related genes

Fig.5 Effects of MitoQ on mitochondrial membrane potential and ATP content of BMSCs A and B represented the mitochondrial membrane potential level (scale bar=100 μm) and ATP content of BMSCs after MitoQ treatment.

References 21

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Effects of MitoQ on mitochondrial function and antioxidant capacity of canine bone marrow mesenchymal stem cells

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