| Research BackgroundAcute spinal cord injury(SCI) is the common and frequently-occurring disease in spinal surgery. Morbidity has been increasing year by year. The injury results in not only dysfunction of sensation and motion and dysfunction of excretion but also dysfunction of reproductive system. The dysfunction of sensation and motion and dysfunction of excretion caused by SCI has been a hot research issue and some progresses have been seen in related treatments. However, researches about dysfunction of reproductive system seem not so popular, with many present reports contradictory and no good treatment found. Based on the discovery home and abroad that sacral nerve electrostimulation can improve the dysfunction of reproductive system, this research uses rats model with Acute SCI, bilateral sacral nerve3electrostimulation as intervention, and observes the effects of sacral nerve electrostimulation on the testis, epididymis, prostate, sperm, Hypothalamus-Pituitary-Gonad Axis Hormone of the rats with SCI, so as to gain further insight into nerves function in reproductive system and provide new basis for clinical treatments on patients with dysfunction of reproductive system caused by SCI.ObjectiveTo observe the effects of sacral nerve3electrostimulation on the rats with SCI, which includes the effect on the rat’s weight and morphology of testis, epididymis and prostate, quantity and quality of sperm as well as Hypothalamus-Pituitary-Gonad Axis Hormone, and provide new ideas for clinical treatments on patients with dysfunction of reproductive system caused by SCI.Methods54SD rats were randomly divided into three groups. The Sham Operation group (SO group n=18) only adopted paravertebral muscle stripping; the Spinal Cord Injury group (SCI group n=18) adopted modified Allen’s weight drop at T10spinal level; the Spinal Cord Injury Electrostimulation group (SCIE group n=18), based on modified Allen’s weight drop at T10spinal level, electrostimulated bilateral sacral nerve3, set at3V,20Hz,10mA and200us, which each time last15s with15s interval,15mins with15mins interval for two hours every day. Six rats from each group were killed during the second, the fourth and the sixth week respectively.(1) Measured the weight of rat’s testis, epididymis and prostate while the pathological changes of testis, prostate and epididymis were examined using Hematoxylin-Eosin (HE)staining, observed through optical microscope. The testis samples were also observed through electron microscope.(2) Collected epididymis sperms and made them into sperm suspension and calculated sperm numbers. Graded the sperms according to WHO sperm activity grading standard, and calculated sperm activity ratio, inactivity ratio and forward-moving ratio. Observed sperms morphology through Iron Hematoxylin staining. (3) Used orbital vein blood sampling and ELISA method to examine Gonadotropin Releasing Hormone, Follicle Stimulating Hormone, Luteinizing Hormone, Testosterone and Androgen Binging Protein in rat’s blood serum.Based on SPSS19.0statistical software, the data between different time points in the same group and that between the same time points in different groups would be analyzed through one-way ANOVA. The effect of time and intervention in different groups would be based on analysis of variance of factorial design. P<0.05was interpreted as significant.Results(1) Effect on Weight of Organ(1.1) Testis Weight:Little change of testis weight in SCI group were observed during the second and fourth week but a considerable decrease of testis weight in SCI group was observed during the sixth week compared to SCIE group (P<0.05); no significant change of testis weight in SCIE group was observed at all time points.(1.2) Epididymis Weight:The weight of epididymis in SCI group was decreasing at the fourth week, significantly lower than that of SO group during the sixth week (P<0.05). A little decrease of epididymis weight in SCIE group was observed but no significant change compared to that in SO group.(1.3) Prostate Weight:No significant change of prostate weight in SCI group was observed during the second and the fourth week, but a significant increase of prostate weight in this group was observed compared to SO group and SCIE group (P<0.05); little change of prostate weight in SCIE group was observed at all time points.(2) Effect on Organ Morphology(2.1) Effect on Testis Morphology:In testis of rats in SCI group under light microscope since the second week, the spermatogenic cells were disorganized, mature sperms in lumen were in decrease, interstitial edema appeared; with time passing on, these changes became more obvious. And until the sixth week, lots of cells of all kinds died and mature sperms in lumen decreased significantly and interstitial edema disappear. Observation through electronic microscope showed that vacuoles appeared in spermatogenic cells and sperm cells in SCI group and with time moving on, more necrotic fragments in cells appeared. However, through light microscope and electronic microscope, a better situation was observed in spermatogenic cells organization, interstitial edema, sperm decrease and cell dying in SCIE group at the same time points compared to that in SCI group.(2.2) Effect on Prostate Morphology:Since the second week, glandular epithelium proliferation, glandular lumens atrophy, interstitial proliferation were becoming more serious as time passing on and these changes became more significant during the fourth and sixth week. But the situation in SCIE group was better than that of SCI group at the same time points.(2.3) Effect on Epididymis Morphology:During the second week, interstitial proliferation, edema, inflammatory cells infiltration, and lumen sperms decrease appeared in epididymis of SCI group; with time passing on, those changes became more obvious. However, the situation of interstitial proliferation, edema, inflammatory cells infiltration, and lumen sperms decrease appeared in rats epididymis of SCIE group was better than that of SCI group at the same time points.(3) Effect on Sperms(3.1) Effect on Sperms Quantity:In SCI group, no significant decrease in sperm number was observed during the second week but a considerable decrease in sperm number appeared during the fourth and sixth week compared to SO group (P<0.05). In SCIE group, sperm number decreased greatly during the fourth and sixth week compared to SO group (P<0.05), but it was considerably higher than that in SCI group (P<0.05).(3.2) Effect on Sperm Quality:In SCI group, during the second week, little change appeared in sperm activity ration and inactivity ration, but during the fourth and sixth week, compared to the SO group, a considerable decrease in sperm activity ratio and a significant increase in sperm inactivity ration appeared (P<0.05). In SCIE group, no significant change appeared in sperm activity ratio and inactivity ratio during the second and fourth week (P<0.05). During the sixth week, ratio activity decreased considerably compared to SO group yet was sharply higher than that in SCI group (P<0.05); sperm inactivity ratio in SCIE group was considerably higher than that in SO group but was considerably lower than that in SCI group (P<0.05). In SCI group, sperm forward moving ratio showed no significant change during the second week, but a considerable decrease during the fourth and sixth week in contrast to SO group (P<0.05). In SCIE group, sperm forward moving ratio showed no significant change during the second and fourth week, but a considerable decrease during the sixth week in contrast to SO group and a considerable increase during the sixth week in contrast to SCI group (P<0.05).(3.3) Effect on Sperm Morphology:In SCI group, during the second week, sperm number decreased and some sperm tail broke; during the fourth week, teratosperm increased with abnormal head sperms appearing and the changes became more obvious during the sixth week. In SCIE group, during the second group, sperm shape was generally normal, the number decreased slightly and abnormal sperms and sperm number decrease happened during the fourth and sixth week, but the situation was better than that in SCI group.(4) Effect on Hypothalamus-Pituitary-Gonad Axis Hormone(4.1) Effect on GnRH:No significant change was observed in GnRH in SCI and SCIE group during the second week. But during the fourth and sixth week, GnRH in SCI group was considerably lower than that in SO group but was considerably higher than that in SCI group (P<0.05).(4.2) Effect on LH:In SCI group, LH was considerably higher than that in SO group during the second, fourth and sixth week. In SCIE group, no striking change in LH was observed during the second group, but a considerable increase of LH appeared during the fourth and sixth week compared to SO group yet it was still significantly lower than that in SCI group (P<0.05).(4.3) Effect on FSH:In SCI group, FSH considerably rose in contrast to SO group during the second, fourth and sixth group (P<0.05). In SCIE group, little change appeared in FSH during the second and fourth week, but a considerable rise was seen in FSH in contrast to SO group during the sixth week, yet the FSH in this group was still lower than that in SCI group (P<0.05).(4.4) Effect on T:In SCI group, no significant change was observed in T during the second week but a considerable decrease in T was observed in contrast to SO group during the fourth and sixth week (P<0.05). In SCIE group, little change was observed in T during the second week but a considerable decrease in T during the fourth and sixth week in contrast to SO group yet a considerable increase in T in contrast to SCI group during the sixth week (P<0.05).(4.5) Effect on ABP:In SCI group, ABP was considerably lower than that in SO group during the second, fourth and sixth week. In SCIE group, ABP was considerably decreasing compared to that in SO group during the second, fourth and sixth group but it was considerably higher than that in SCI group during the fourth and sixth week (P<0.05).Conclusions(1) Acute SCI would result in male rats spermatogenic cells death, decrease in sperm quantity and quality, epididymis interstitial edema and proliferation, prostate mesenchyme and epithelium proliferation, glandular lumens atrophy, weight loss in testis and epididymis, weight gaining in prostate, GnRH, T and ABP going down while LH and FSH going up.(2) Bilateral sacral nerve3electrostimulation during spinal cord injury would depress weight loss in rats testis and epididymis and weight gaining in prostate, reduce spermatogenic cells death, increase sperm quantity and improve sperm quality and ease glandular epithelium proliferation and glandular lumens atrophy, to some extent HPGA dysfunction caused by SCI could be improved.(3) A new direction for clinical treatments on patients with dysfunction of reproductive system caused by SCI would be provided. |
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