Experimental Research Of Influence Of Calcitonin On Infertility Human Sperm Function In Vitro

Objective:This study investigated the influence of salmon calcitonin(sCT)of different concentrations on human sperm motility function,integrity of sperm membrane,capacitation and acrosome reaction from infertility patients in vitro.This provides helps for male infertility diagnosis,treatment in vitro and artificial insemination of assisted reproductive techniques(TRT).Methods:This study includes four parts.Sperm were aseptically obtained by masturbation and prepared by swim-up technique to produce a spermatozoa solution of high motility from 35 infertile men.1.The spermatozoa solution was incubated with the different concentrations of calcitionin at 37℃in vitro.Motive parameters were measured by computer-aided sperm analysis (CASA).Measurements were carried out at 0min,30 min and 60 min in 15specimens,including Sperm motility,Sperm viability and Sperm parameter.2.The hypo-osmotic swelling(HOS) test was used to assess the sperm function.3.The incapacitated spermatozoa solution was incubated with the different concentrations of calcitionin at 37℃in vitro and was promoted to capacitation.After capacitation the induced acrosome reaction(IAR)by progesterone was determined by means of a flow cytometer.The FITC-PSA was used to stain the sperm acrosome,which can radiate green fluorescence at 510～550nm wavelength as excited by 488nm wavelength laser.The value of FITC fluorescence was accumulated and calculated by a special computer as the stained sample passed the flow cytometer.4.The incapacitated spermatozoa solution added fucose was incubated at 37℃for 4 hours in vitro and was promoted to capacitation.After capacitation the different concentrations of calcitonin was added to sample tube.The spontaneous acrosome reaction(SAR)affectded by different concentrations of calcitonin was measured by ways of flow cytometer.Results:1)a The spermatozoa incubated with salmon calcitionin of 0.5nmol/L and 5nmol/L in vitro showed significantly higher on Sperm motility,Sperm viability and Sperm motive parameters than the control group at 30min.(p＜0.01).And 0.05nmol/L sCT group did not show significantly different than the control group.There were not significant difference among the rest groups.b Salmon calcitonin of 5nmol/L did not show significantly different in different periods after incubation.2)The spermatozoa incubated with different concentrations of salmon calcitonin in vitro showed significant difference than the control group.3)The values of FITC fluorescence of 0.05nmol/L,0.5nmol/L,5nmol/L groups and progesterone group in vitro showed significantly lower than the control group.The difference showed significant (P＜0.01).The values of FITC fluorescence of 0.5nmol/L and 5nmol/L groups showed significantly lower than progesterone group.The difference showed significant(P＜0.05).The values of FITC fluorescence of 0.05nmol/L group showed lower than progesterone group,but the difference did not show significant(P＞0.05).The values of FITC fluorescence of 0.5nmol/L and 5nmol/L groups did not show significantly different.4)The values of FITC fluorescence of 0.5nmol/L and 5nmol/L groups in vitro showed significantly higher than the control group.The difference showed significant(P＜0.01).The value of FITC fluorescence of 0.05nmol/L group showed higher than the control group,but the difference did not show significant(P＞0.05).The value of FITC fluorescence of 5nmol/Lgroup in vitro showed higher than 0.5nmol/L group. The difference also did not showed significant(P＞0.05).Conclusion:Calcitonin can obviously improve human sperm motility function.Calcitonin does not affect sperm hypo-osmotic swelling rate in vitro.For the incapacitated spermatozoa solution calcitonin would stimulate capacitation and enhance the induced acrosome reaction(IAR)by progesterone;but after capacitation calcitonin would begin to inhibit spontaneous acrosome reactions.The responses obtained in vitro suggest that this peptide could also be used effectively in infertility research and clinic.