Calis P, Bozdag G, Karakoc Sokmensuer L, Kender N
Department of Obstetrics and Gynecology, Hacettepe University School of Medicine, Ankara, Turkey.
Department of Obstetrics and Gynecology, Hacettepe University School of Medicine, Ankara, Turkey.
Eur J Obstet Gynecol Reprod Biol. 2015 Feb;185:126-30. doi: 10.1016/j.ejogrb.2014.12.006. Epub 2014 Dec 23.
OBJECTIVE(S): To evaluate the effect of ischemia-reperfusion on follicle count and viability of follicles in a rat model with adnexal torsion.
A total of 24 female adult Wistar albino rats were included in the study. In the first laparotomy, right ovaries were twisted at 720° in the counterclockwise for a duration of two (n=12), four (n=6) or 16 hours (n=6), whereas contralateral sites were kept as controls. In the second laparotomy, the right ovaries were detorsioned. Twenty-eight days after the detorsion, both ovaries were retrieved with the third laparotomy and placed in formaldehyde for subsequent examination under light microscopy and immunohistochemistry for proliferating cell nuclear antigen (PCNA). Besides histological evaluation by only using a standard scoring system, the viability of the follicles was also assessed by PCNA with immunohistochemical staining.
Macroscopically, after two, four and 16 hours of torsion, all the twisted ovaries had enlarged and demonstrated a dark, dusky appearance. However, under light microscopy with 10X magnification, the mean primordial+primary follicle count was comparable in twisted and control sides (26.7±7.6 vs. 28.1±5.9 respectively). In terms of the means of total tissue damage, only the loss of cohesion was found to be significantly different compared to the control sides (2.78±0.86 vs. 2.17±0.86, p=0.017). Immunohistochemical staining revealed significantly lower PCNA counts in the 16-hour torsion group only.
CONCLUSION(S): Even though ovarian reserves, reflected by primordial and primary follicle count, are similar between torsioned and contralateral ovaries, the viability of the remaining follicles might be affected by a long-standing ischemia-reperfusion phenomenon. Follicle count may reflect an idea of fertility potential, but immunohistochemical staining using viability markers such as PCNA confirms the viability of the counted follicles and appears to be a more precise approach necessary for demonstrating the functional status.
评估缺血再灌注对附件扭转大鼠模型中卵泡数量及卵泡活力的影响。
本研究共纳入24只成年雌性Wistar白化大鼠。在首次剖腹手术中，右侧卵巢逆时针扭转720°，持续时间分别为2小时（n = 12）、4小时（n = 6）或16小时（n = 6），对侧作为对照。在第二次剖腹手术中，解除右侧卵巢扭转。扭转解除28天后，通过第三次剖腹手术取出双侧卵巢，置于甲醛中，随后进行光镜检查及增殖细胞核抗原（PCNA）免疫组化检查。除仅使用标准评分系统进行组织学评估外，还通过PCNA免疫组化染色评估卵泡活力。
宏观上，扭转2小时、4小时和16小时后，所有扭转的卵巢均肿大，外观呈暗黑色。然而，在10倍放大的光镜下，扭转侧和对照侧的原始卵泡 + 初级卵泡平均数量相当（分别为26.7±7.6和28.1±5.9）。在总组织损伤方面，仅发现粘连丧失与对照侧相比有显著差异（2.78±0.86对2.17±0.86，p = 0.017）。免疫组化染色显示仅16小时扭转组的PCNA计数显著降低。
尽管原始卵泡和初级卵泡数量反映的卵巢储备在扭转侧和对侧卵巢之间相似，但长期的缺血再灌注现象可能会影响剩余卵泡的活力。卵泡数量可能反映生育潜力，但使用PCNA等活力标志物进行免疫组化染色可确认所计数卵泡的活力，似乎是证明功能状态所需的更精确方法。
