抗FSH自身抗体对大鼠睾丸生精功能影响的初步研究

目的:建立抗FSH自身抗体大鼠模型,同时研究抗FSH自身抗体对雄性大鼠生精功能的影响。方法:30只SD大鼠(21d龄),随机分为实验组、对照组,各15只。合成大鼠FSHβ亚基上一段特异性氨基酸序列(18肽),将合成多肽与钥孔戚血蓝素(keyholelimpethemocyanin,KLH)偶联,免疫SD大鼠,作为实验组。对照组大鼠用KLH免疫。初次免疫之后,每隔2周加强免疫1次,共7次,于第3次加强免疫后1周(即,免疫第49d)眼眶取血测定血清抗体效价,之后每隔两周测一次抗体效价,至实验结束。并于免疫第77d、91d和105d时,分别处死实验组和对照组大鼠各5只,用光镜和电镜观察大鼠睾丸生精小管结构和附睾精子结构的变化,计数附睾尾精子数量及肿胀精子百分率,并用ELISA法检测大鼠血清T水平。结果:免疫第49d后,实验组大鼠血清抗18肽-KLH抗体效价为1∶200,免疫第63d后,抗体效价达到1∶400,之后抗体效价一直维持在1∶400。免疫第91d后,实验组大鼠附睾尾肿胀精子百分率显著低于对照组(60.4±6.23vs50.60±3.05,P<0.05);免疫第105d后,实验组大鼠生精小管内生精细胞数量及管腔内精子数量均减少,且附睾尾精子数量(46.08±6.56vs32.53±3.41)和肿胀精子百分率(60.60±5.86vs48.60±3.85)均显著低于对照组(P<0.05),而血清T水平显著高于对照组(P<0.05)。结论:抗FSH自身抗体可能会引起睾丸生精障碍。

Primitive Research on the Effect of FSH Autoantibody on the Spermatogenic Capability of the Rat Testis

OBJECTIVE: To establish rat models of FSH autoantibody and to investigate the effect of FSH autoantibody on the spermatogenic capability of rat testis. METHODS: Thirsty 21-day old SD rats were randomly divided into an experimental and a control group of equal number. A specific polypeptide corresponding to the rat FSHbeta subunit was synthesized and coupled to (keyhole limpet hemocyanin) KLH. The rats in the experimental group were immunized with polypeptide-KLH and these in the control group with KLH. Further immunization was performed every 2 weeks for 7 times. On the 77th, 91st and 105th day of the immunization, 5 rats from the experimental group and another 5 from the control group were killed. Then the structures of the seminiferous tubule and epididymal sperm were observed by light and electron microscope, respectively. Meanwhile, the counts of sperms and the percentage of swelled sperm were calculated. And the level of serum testosterone was detected by enzyme-linked immunospecific assay (ELISA). RESULTS: The titer of the anti-polypeptide antibody was 1:200 on the 49th day of the immunization, and reached 1:400 on the 63rd. Compared with the control group, the percentage of swelled sperm significantly decreased on the 91st day (60.4 +/- 6.23 vs 50.60 +/- 3.05, P < 0.05), and the number of spermatogenic cells and sperms in seminiferous tubules reduced on the 105th day in the experimental group, the counts of sperms (46.08 +/- 6.56 vs 32.53 +/- 3.41) and the percentage of swelled sperm (60.60 +/- 5.86 vs 48.60 +/- 3.85) significantly lower (P < 0.05), while the level of serum T significantly higher than that in the control group (P < 0.05). CONCLUSION: FSH autoantibody might cause testis dyszoospermia.