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. 2018 May 15;8(1):7603.
doi: 10.1038/s41598-018-25687-z.

Resveratrol significantly improves the fertilisation capacity of bovine sex-sorted semen by inhibiting apoptosis and lipid peroxidation

Chong-Yang Li  1 , Ya-Han Zhao  1 , Hai-Sheng Hao  1 , Hao-Yu Wang  1 , Jin-Ming Huang  2 , Chang-Liang Yan  3 , Wei-Hua Du  1 , Yun-Wei Pang  1 , Pei-Pei Zhang  1 , Yan Liu  1 , Hua-Bin Zhu  4 , Xue-Ming Zhao  5
Affiliations

Resveratrol significantly improves the fertilisation capacity of bovine sex-sorted semen by inhibiting apoptosis and lipid peroxidation

Chong-Yang Li et al. Sci Rep. .

Abstract

The aim of this study was to test the effects of five different concentrations (0, 10-3, 10-4, 10-5, and 10-6 M) of resveratrol (Res) supplementation in bull sperm washing and fertilisation medium on levels of reactive oxygen species (ROS), phosphatidylserine (PS) externalisation, mitochondrial membrane potential (Δψm), ATP and malondialdehyde (MDA), acrosomal integrity, blastocyst rate, and blastocyst quality after in vitro fertilisation (IVF). The results for sex-sorted sperm from three bulls showed: (1) ROS and MDA levels in 10-3 M and 10-4 M Res groups were significantly lower than those of controls (P < 0.05); (2) the percentage of viable sperm, percentage of sperm with high Δψm, and the ATP content in 10-3 M and 10-4 M Res groups were significantly higher than those of controls (P < 0.05); (3) the percentage of viable sperm with acrosomal integrity, and the blastocyst percentage and quality of the 10-4 M Res group were significantly higher than those of controls (P < 0.05). In conclusion, 10-4 M Res supplementation in washing and fertilisation medium of sex-sorted bull sperm significantly decreased ROS, PS externalisation, and MDA, and protected mitochondrial function and acrosomal integrity, thereby increasing blastocyst percentage and quality following IVF.

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Conflict of interest statement

The authors declare no competing interests.

Figures

Figure 1
Figure 1
Effect of Res on ROS levels in sex-sorted bull sperm. (A) Representative images showing ROS staining. Scale bar = 20 μm. (B) a,b,cValues with no common superscript are different (P < 0.05).
Figure 2
Figure 2
Effect of Res on PS externalisation in sex-sorted bull sperm. (A) PS externalisation assay. (a) Negative control. (b) Annexin V FITC staining control. (c) PI staining control. (d) Analysis of sex-sorted bull sperm. Quadrants represent viable sperm (lower-left quadrant), necrotic sperm (upper-left quadrant), early apoptotic sperm (lower-right quadrant), and early necrotic sperm (upper-right quadrant). (B) a,b,c,d,eValues with no common superscript are different (P < 0.05).
Figure 3
Figure 3
Effect of Res on Δψm in sex-sorted bull sperm membranes. (A) Δψm assays. (a) Negative control. (b) Positive control. (c) Analysis of sex-sorted bull sperm. Quadrants represent sperm with low Δψm (lower-right quadrant) and high Δψm (upper-right quadrant). (B) a,b,c,dValues with no common superscript are different (P < 0.05).
Figure 4
Figure 4
Effect of Res on ATP content in sex-sorted bull sperm. a,b,c,dValues with no common superscript are different (P < 0.05).
Figure 5
Figure 5
Effect of Res on MDA content in sex-sorted bull sperm. a,b,c,dValues with no common superscript are different (P < 0.05).
Figure 6
Figure 6
Representative images of acrosome staining of sex-sorted bull sperm. S1, viable sperm with integral acrosomes (PI−/PNA−). S2, viable sperm with damaged acrosomes (PI−/PNA+). S3, dead sperm with acrosomal integrity (PI+/PNA−). S4, dead sperm with damaged acrosomes (PI+/PNA+). Scale bar = 20 μm.
Figure 7
Figure 7
Effect of Res on acrosome integrity in sex-sorted bull sperm. a,b,cValues with no common superscript are different (P < 0.05).
Figure 8
Figure 8
Effect of Res on IVF efficiency in sex-sorted bull sperm. a,b,c,dValues with no common superscript are different (P < 0.05).
Figure 9
Figure 9
Effect of Res on the expression of BAX and BCL2L1 mRNAs in bovine blastocysts from sex-sorted bull sperm. a,bValues with no common superscript are different (P < 0.05).
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