Taylor U, Schuberth H J, Rath D, Michelmann H W, Sauter-Louis C, Zerbe H
Institute of Animal Breeding, Mariensee (FAL), Neustadt, Germany.
Reprod Domest Anim. 2009 Apr;44(2):180-8. doi: 10.1111/j.1439-0531.2007.01015.x. Epub 2008 Sep 27.
A post-breeding migration of leucocytes (PMN) into the uterus is considered to be an important reason for sperm losses. Minimizing such effects may be necessary for successful insemination with low sperm numbers, as required with sex-sorted spermatozoa. We examined the magnitude of PMN influx 3 h after pre- or post-ovulatory insemination with various combinations of seminal plasma (SP), semen extender Androhep (AH; Minitüb, Tiefenbach, Germany) and sperm preparations (S). Pre-ovulatory inseminations with preparations containing 98% AH caused a massive influx of PMN, independent of whether spermatozoa were present (628 +/- 189 x 10(6) leucocytes/uterine horn) or not (580 +/- 153 x 10(6)). Post-ovulatory, 98% AH caused a comparable immigration only in the absence of sperm cells (AH: 569 +/- 198 x 10(6), AH+S: 162 +/- 102 x 10(6)). The presence of SP significantly dampened the numbers of recruited uterine leucocytes. The reaction to all inseminates containing 98% SP both with and without spermatozoa, used before ovulation (SP: 14 +/- 6 x 10(6), SP+S: 73 +/- 27 x 10(6)) and after ovulation (SP: 60 +/- 32 x 10(6), SP+S: 51 +/- 33 x 10(6)) did not differ significantly from controls using phosphate buffered saline (PBS) (pre-ovulatory: 1 +/- 1 x 10(6), post-ovulatory: 11 +/- 9 x 10(6)). Quantitative in vitro transmigration assays with blood-derived PMN proved that AH-induced leucocyte migration into the uterus to be not as a result of direct chemotaxis, because, on account of the chelator citrate, AH significantly inhibited the transmigration towards recombinant human Interleukin-8 (rhCXCL8) (AH: 14 +/- 5% migration rate vs controls: 37 +/- 6%, p < 0.05). Supernatants of spermatozoa incubated in PBS for 1, 12 or 24 h showed neither chemoattractive nor chemotaxis-inhibiting properties. SP at > or =0.1% [v/v] significantly inhibited the in vitro transmigration of PMN. With respect to in vivo migration of neutrophils, the striking difference in the results between semen extender and seminal plasma suggests that adaptation of extender composition is needed to reflect more closely the in vivo regulatory potential of natural seminal plasma.
白细胞（多形核白细胞，PMN）在配种后的子宫内迁移被认为是精子损失的一个重要原因。对于使用低精子数量进行成功授精（如性别分选精子所要求的那样），尽量减少这种影响可能是必要的。我们研究了在排卵前或排卵后授精3小时后，精液血浆（SP）、精液稀释剂Androhep（AH；德国蒂芬巴赫的Minitüb公司）和精子制剂（S）的各种组合导致的PMN流入量。使用含有98%AH的制剂进行排卵前授精会导致PMN大量流入，无论是否存在精子（628±189×10⁶白细胞/子宫角）（580±153×10⁶）。排卵后，98%AH仅在不存在精子细胞时导致类似的迁移（AH：569±198×10⁶，AH+S：162±102×10⁶）。SP的存在显著抑制了募集到的子宫白细胞数量。对排卵前（SP：14±6×10⁶，SP+S：73±27×10⁶）和排卵后（SP：60±32×10⁶，SP+S：51±33×10⁶）使用的所有含有98%SP且有无精子的授精剂的反应与使用磷酸盐缓冲盐水（PBS）的对照组（排卵前：1±1×10⁶，排卵后：11±9×10⁶）没有显著差异。用源自血液的PMN进行的定量体外迁移试验证明，AH诱导的白细胞向子宫内迁移不是直接趋化作用的结果，因为由于螯合剂柠檬酸盐，AH显著抑制了向重组人白细胞介素-8（rhCXCL8）的迁移（AH：迁移率14±5%，对照组：37±6%，p<0.05）。在PBS中孵育1、12或24小时的精子上清液既没有趋化吸引特性也没有趋化抑制特性。≥0.1%[v/v]的SP显著抑制PMN的体外迁移。关于中性粒细胞的体内迁移，精液稀释剂和精液血浆之间结果的显著差异表明，需要调整稀释剂的成分，以更紧密地反映天然精液血浆的体内调节潜力。
