冷冻保存对人类精子膜完整性的影响

Effect of cryopreservation on human sperm membrane integrity

本文建立“低渗肿胀-伊红”（HOS－EY）结合试验探讨冷冻保存对人类精子头部和尾部膜完整性的影响。HOS－EY试验按照精子尾部肿胀与否和头部着色与否区分为4种膜类型。冷冻保存后，头膜一尾膜均完整的Ⅳ型精子率和头膜一尾膜均损伤的Ⅰ型精子率分别显著减少和增多（n＝50，P＜0．001），表明冷冻一解冻过程对精子膜完整性有严重影响；头膜损伤-尾膜完整的Ⅲ型精子率在冷冻组显著增多，提示精子头部或尾部的冷冻损伤是独立发生的，头膜损伤易于尾膜；头膜完整-尾膜损伤的Ⅱ型精子在冷冻组的发生率低；冷冻精子Ⅳ型精子率与冷冻精子活动率呈显著正相关（n＝50，r＝0．70，P＜0.001）,但与冷冻精子空卵率不呈相关性（n＝25，r=0。22，P＞0.05）。

The hypoosmotic swelling test and eosin Y exelusion were combined to form a single test(HOS - EY test)to evaluate the effect of cryopreservation on Plasma membrme integrity in the head and tail of human sperm. This combined test can identify four four of membrane integrity according to the morphologi cal changes in membranes at both the head and tail regions of the sperm. After cryopreservation (n =50), there was a marked decline in the Percentage of type IV(head rnelnblane intact/tail membrane intact), and a significant increase in percentages of types Ⅰ andⅡ (head membrane damage /tail membrane damage; head membrane damage/tail membrane intact). The value of typeⅢ (a transition state)had a wide range of varieties,whereas type Ⅱ (head membrane intacut/tail membrane damage)was uncommon finding after thawing. A high correlation was observed betWeen the percentage of type Ⅳ and motility in cryopreserved sperm(n = 50, r = 0.74, P < 0.001 ). However, this percentage did not cormlate with the percentage of cryopreserved sperm penehalion(n = 25, r = 0. 22, P > 0.05). It is concluded that: (1 )The HOS - EY test has complelnentaly adventages for shawing four Patterns of membranes inthoty of individual sperm; (2)Cryopreservation causes a signifi cant membrane rupture in the head and tall regions of the sperm; (3)Damage of the head and tail occured independently,type Ⅲ was a main transition state of membrane cryodamage; (4) Cryopreserved sperm with intact membranes was closely related to post - thaw polity, but not to feltilizing potential.