Affect of Trichomonas vaginalis excretory secretory protein on sperm quality of mice

doi:10.12140/j.issn.1000-7423.2022.02.016

Abstract

Abstract:

Trichomonas vaginalis excretory secretory protein (TvESP) was collected for determining the concentration using Bradford method kit and further analyzed of TvESP by thesodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Mouse sperm were collected from the epididymis of Kunming mice, and the sperm density was adjusted to 1 × 10⁷/ml. Sperm were divided into 6 groups and co-cultured with TvESP at final concentrations of 0, 5, 10, 20, 40 and 80 ng/μl, respectively. After co-culturing with TvESP for 1 h, sperm motility was observed using an inverted microscope, and sperm apoptosis was detected by flow cytometry. After co-culturing for 0.5, 1 and 2 h, the sperm viability was analyzed by trypan blue staining. After sperm were incubated with TvESP at final concentrations of 0 and 40 ng/μl for 1 h, sperm were stained with a fluorescent dye and the acrosome was observed under a fluorescence microscope. One-way ANOVA was used for comparing between groups. The results showed that the percentage of active sperm in the 5, 10, 20, 40 and 80 ng/μl groups were (31.90 ± 4.10)%, (23.00 ± 4.90)%, (18.80 ± 3.50)%, (13.60 ± 1.20)% and (8.60 ± 1.20)%, respectively, which were significantly lower than that of the 0 ng/μl concentration group (44.50 ± 3.20)% (F = 30.76, P < 0.01). The apoptosis rates of sperm in the 5, 10, 20, 40 and 80 ng/μl groups were (17.99 ± 1.73)%, (22.43 ± 1.53)%, (26.76 ± 2.20)%, (32.05 ± 1.68)% and (41.37 ± 2.28)%, respectively, which were significantly higher than that of the 0 ng/μl group (15.42 ± 1.10)% (F = 256.79, P < 0.01). After treatment with TvESP for 0.5 h, the mortality rates of sperm treated with 10 ng/μl (11.90 ± 1.70)%, 20 ng/μl (14.10 ± 1.40)%, 40 ng/μl (16.30 ± 1.20)%, 80 ng/μl (17.90 ± 1.30)% of TvESP were significantly higher than that of the 0 ng/μl group (7.80 ± 1.10)% (F = 15.72, P < 0.05). The sperm mortality rate in the 5 ng/μl group (10.10 ± 1.40)% was not statistically different from that of the 0 ng/μl group. After treatment with TvESP for 1 h, the sperm mortality rates in the 5, 10, 20, 40 and 80 ng/μl groups were (12.90 ± 0.90)%, (14.60 ± 0.90)%, (16.20 ± 1.30)%, (18.10 ± 1.10)%, (20.00 ± 1.40)%, which were significantly higher than that of the 0 ng/μl group (9.90 ± 1.90)% (F = 15.76, P < 0.01). After 2 h treatment, the mortality rates in the 10, 20, 40 and 80 ng/μl groups were (17.00 ± 1.20)%, (18.30 ± 1.20)%, (20.50 ± 1.20)% and (22.80 ± 1.50)%, respectively, which were significantly higher than that of the 0 ng/μl group (12.90 ± 0.50)% (F = 22.22, P < 0.01), and the mortality rate in the 5 ng/μl group (15.00 ± 0.40)% was not statistically different from that of the 0 ng/μl group. After co-culturing the sperm with TvESP at a final concentration of 40 ng/μl for 1 h, the integrity of acrosome was destroyed. All results suggested that TvESP can reduce sperm quality.

Key words: Trichomonas vaginalis, Secretory protein, Mouse, Sperm quality

CLC Number:

R383.211

LUO Cheng-yang, HAO Li-xia, GUO Li-hua, WANG Bing-li, JI Si-fan, LIAN An-na, ZHU Xiao-hui, MEI Xue-fang, TIAN Xiao-wei, WANG Shuai, ZHANG Zhen-chao. Affect of Trichomonas vaginalis excretory secretory protein on sperm quality of mice[J]. CHINESE JOURNAL OF PARASITOLOGY AND PARASITIC DISEASES, 2022, 40(2): 236-241.

Figures/Tables 5

References 25

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