宫腔粘连动物模型的建立

Establishment of an animal model of intrauterine adhesions

目的建立稳定的宫腔粘连(IUA)病理性再生动物实验模型。方法将30只雌性SD大鼠随机分为实验组和对照组,每组各15只。实验组通过自制2 mm直径刮匙分别对动情期大鼠的左、右侧子宫内膜进行轻度、重度机械损伤并设为轻度机械损伤组、重度机械损伤组;对照组大鼠开腹后不做子宫内膜损伤处理,进行假手术对照。分别于术后0 h、24 h、72 h、5 d、7 d从每组随机选取3只大鼠取出双侧子宫行快速冰冻切片,分别进行子宫内膜细胞中主要组织相容性复合体Ⅱ类分子(MHCⅡ)、5-溴脱氧尿苷(BrdU)免疫组织化学染色,观察术后7 d不同大鼠内膜腺体情况以及术后不同时期不同机械损伤程度的大鼠子宫内膜组织炎性反应及子宫内膜组织增生修复变化。结果轻度机械损伤组术后7 d可见少量红染含BrdU的细胞及蓝染的MHCⅡ细胞,子宫内膜基本修复,轻度炎症反应存在;重度机械损伤组术后7 d未见腺体或腺体数目极少,未见明显红染含BrdU的细胞,可见少许蓝染的MHCⅡ细胞、轻度炎症反应、内膜组织纤维化,局部管腔闭合,出现粘连。与对照组相比,术后7 d大鼠子宫内膜腺体数目随着子宫内膜机械性受损伤程度增加而相应减少(P<0.001)。结论采用切开缝合子宫内膜机械损伤法可以成功建立大鼠IUA模型。大鼠子宫内膜轻度机械损伤后可以再生达到基本修复水平,而重度机械损伤后子宫内膜组织无法达到基本修复,且出现组织纤维化导致IUA。

Objective To establish a stable animal model for the study of pathological regeneration of intrauterine adhesions(IUAs).Methods Thirty female SD rats were randomly divided into the experimental(n=15)and control groups(n=15).In the experimental group,a self-made 2-mm diameter curette was utilized to create mild and severe mechanical injury to the left and right endometrium of estrous rats and then divided into the mild and severe mechanical injury groups,respectively.In the control group,the rats were not treated with endometrial injury after open surgery,which were used as sham surgery controls.At 0 h,24 h,72 h,5 d,and 7 d after surgery,bilateral uteruses were removed from 3 rats in each group and prepared for quick frozen sections for MHCⅡ and BrdU immunohistochemical staining.The endometrial glands of different rats were observed at postoperative 7 d,and the inflammatory reaction of endometrial tissues and the changes of endometrial tissue proliferation and repair were also observed in rats with different degrees of mechanical injury at different time points after surgery.Results In the mild mechanical injury group,a small amount of red-stained BrdU-containing cells and blue-stained MHCⅡ cells were seen at 7 d after surgery,and the endometrium was basically repaired,while mild inflammatory reaction occurred.In the severe mechanical injury group,no endometrial glands or very few glands were detected,no obvious red-stained BrdU-containing cells were seen,a small quantity of blue-stained MHCⅡ cells,mild inflammatory reaction,endometrial fibrosis,local lumen closure and adhesion were observed at postoperative 7 d.Compared with the control group,the number of endometrial glands in the rat uterine cavity was decreased along with the increase in the degree of mechanical endometrial injury at 7 d after surgery,and the difference was statistically significant(P<0.001).Conclusions The rat model of IUAs can be successfully established by incision and suture of endometrial mechanical injury method.After mild mechanical injury,the endometrium of rats can regenerate and achieve basic repair.Nevertheless,the endometrial tissues cannot achieve basic repair after severe mechanical injury.In addition,tissue fibrosis occurs,leading to IUAs.