|
Objective: Determination of normal fertility and weak sperm semen of patients with L-carnitine (L-carnitine) content to study different concentrations of L-carnitine in vitro on human sperm vitality, viability, motor function, function of sperm membrane integrity, acrosome reaction of L-carnitine on human sperm, to evaluate the effect of the treatment of male infertility. Methods: health, has normal fertility in male 20 cases, infertility patients, 20 cases of azoospermia by routine examination of semen weakness. Masturbation semen. The first part: the use of high performance liquid chromatography determination of the content of L-carnitine in the two sets of samples of semen. Part II: the final concentration for 4mmol / L, 8mmol / L and 12 mmol / L of L-carnitine in vitro by upstream optimization process two groups of male sperm were incubated, and the establishment of the normal control group, in 0min, 30min, 60min, 120min when they were semen analysis. Part III: the second portion of each group of 60 minutes of semen were hypotonic swelling test (HOS). Part IV: Two groups of male sperm by Raw optimize treatment to induce sperm capacitation, the final concentration for 4mmol / L, 8mmol / L, 12 mmol / L of L-carnitine with capacitated spermatozoa incubated for 60 minutes at the same time established normal control group, and then induce sperm acrosome reaction, labeled with specific fluorescent markers - complexation with fluorescein isothiocyanate pea lectin (FITC-PSA) acrosome, the acrosome complete by flow cytometry sex. Results: 1) The first part, determined by high performance liquid chromatography instrument, normal fertility group with weak sperm seminal plasma L-carnitine concentrations were: (205.67 soil 11.04) mol / L, (112.24 ± 10.27) micromol / L. The significant difference between the two groups was statistically significant (P lt; 0.01). 2) The second part, compared with the control group, normal fertility group with different concentrations of L-carnitine, sperm motility, vitality, and its motion parameters there was no significant change was not statistically significant. However, under the same incubation conditions, can significantly slow down the trend of decline in sperm motility, vitality and VSL, VCL and VAP motion parameters. Weak sperm group under different concentrations of L-carnitine, its vitality, live rate has improved significantly, and the difference was statistically significant (P <0.05) and the curve velocity (VCL), straight-line velocity (VSL), average path velocity (VAP), moving average angle (MAD) side of swing (ALH), swing (WOB) were significantly different (P <0.05); compared with the control group, 4 mmol / L, 8mmol / L, 12 mmol / L group whipping frequency (BCF), linear (LIN) and isotropic (STR) no significant difference (P> 0.05). Show that with the elevated concentration of L-carnitine and the extension of the incubation time, sperm motility, viability, sperm motility was able to maintain a high activity trend. 3) The third part, in the normal fertile men group, the control group, 4mmol / L, 8mmol / L, 12 mmol / L group sperm g-shaped sperm tail swelling rate was no significant difference (P> 0.05). Shows that with the rise of the concentration of L-carnitine, L-carnitine has no effect on the function of sperm membrane integrity of normal fertile men. Different concentrations of L-carnitine in patients with asthenospermia semen, compared with the control group, the 4mmol / L groups, 8mmol / L group, 12 mmol / L group sperm tail swelling rate, g sperm percentage reduced compared with the obvious, the gap statistically significant (P lt; 0.05), show that after the role of L-carnitine, can improve the weak sperm membrane integrity. 4) The fourth part in the normal fertile group, the role of different concentrations of L-carnitine, among the groups acrosome labeling fluorescent value did not differ significantly, in asthenospermia group, a final concentration of 12 mmol / L group sperm acrosome Mark fluorescence significantly higher than that in the control group and the other groups, the difference was significant (P <0.05). Control group, no significant difference (P> 0.05) between the 4mmol / L group, 8mmol / L group. By the concentration of 12 mmol / L L-carnitine incubation, to better promote progesterone-induced acrosome reaction. Conclusion: In patients with asthenospermia semen L-carnitine content than normal fertile men. Sperm in vitro, the proper concentration of L-carnitine can promote human sperm motility and viability rate, improve sperm motility; did not affect the normal function of sperm membrane integrity, but can improve of patients with asthenospermia sperm membrane function; appropriate concentration of L-carnitine can promote the progesterone-induced acrosome reaction, improve the ability of sperm to conceive. The development of sperm in vitro processing technology provides hands-on experience; provide a theoretical basis and theoretical reference for the development of artificial insemination and in vitro fertilization technology.
|