新鲜羊膜对兔宫腔黏连的预防作用及对子宫内膜容受性的改善作用

doi:10.13481/j.1671-587X.20210424

摘要/Abstract

摘要： 目的

建立兔宫腔黏连（IUA）模型，探讨宫腔内放置新鲜羊膜预防IUA的效果和对子宫内膜容受性的改善作用。

方法

雌性新西兰大白兔30只随机分为对照组、模型组和治疗组，每组10只，模型组和治疗组兔用刮勺搔刮宫腔机械性损伤子宫内膜并于宫腔内放置浸泡脂多糖（LPS）的棉线使宫腔感染，建立兔IUA模型，治疗组兔于模型建立同时宫腔内放置新鲜羊膜，对照组为假手术组，不进行宫腔处理。收集建模后第1和2周时的子宫标本，采用HE染色和Masson染色检测各组兔子宫内膜组织中腺体数量和子宫内膜纤维化面积百分率，采用免疫组织化学染色法检测各组兔子宫内膜组织中整合素αVβ3和白血病抑制因子（LIF）蛋白表达水平。

结果

与对照组比较，第1和2周时模型组和治疗组兔子宫内膜组织中腺体数量明显减少（P<0.05），子宫内膜纤维化面积百分率明显升高（P<0.05），子宫内膜组织中整合素αVβ3和LIF蛋白表达水平明显降低（P<0.05）；与模型组比较，第1和2周时治疗组兔子宫内膜组织中腺体数量明显增加（P<0.05），子宫内膜纤维化面积百分率明显降低（P<0.05），子宫内膜组织中整合素αVβ3和LIF蛋白表达水平明显升高（P<0.05）；与第1周时比较，第2周时对照组和治疗组兔子宫内膜组织中腺体数量、子宫内膜纤维化面积百分率及子宫内膜组织中整合素αVβ3和LIF蛋白表达水平差异均无统计学意义（P>0.05），但第2周时模型组兔子宫内膜纤维化面积百分率明显降低（P<0.01），子宫内膜组织中整合素αVβ3和LIF蛋白表达水平明显升高（P<0.05）。

结论

宫腔内放置新鲜羊膜可以减轻IUA的程度，改善子宫内膜容受性，且起效快并作用持久。

关键词: 宫腔黏连, 新鲜羊膜, 整合素αVβ3, 白血病抑制因子, 子宫内膜容受性

Abstract: Objective

To set up the rabbit intrauterine adhesion （IUA） models， and to explore the effect of placing fresh amniotic membrane in the uterine cavity in the prevention of IUA and its improvement effect on the endometrial receptivity.

Methods

Thirty female New Zealand white rabbits were randomly divided into control group， model group and treatment group， with 10 in each group. Using scraping intrauterine mechanical damage to the endometrium and placing cotton lines soaked in lipid polyaccharides （LPS） in the intrauterine cavity， the rabbit IUA models were established in model group and treatment group. For the rabbits in treatment group， fresh amniotic membrane was placed in the uterine cavity at the same time.The control group was a sham operation group with no uterine treatment. The uterine specimens were collected at the 1st and 2nd weeks. The number of glands and the percentages of endometrial fibrosis areas in endometrium tissue of the rats rabbits in various groups were compared by HE and Masson staining， and the expression levels of integrin αVβ3 and leukemia inhibitory （LIF） proteins in endometrium tissue of the rabbits were detected by immunohistochemical staining.

Results

Compared with control group， the number of glands in endometrium tissue of the rabbits in model group and treatment group were decreased significantly at the lst and 2nd weeks （P<0.05），the percentages of endometrial fibrosis areas were increased significantly（P<0.05），and the expression levels of integrin αVβ3 and LIF proteins in endometial tissue were decreased significantly （P<0.05）.Compared with model group， the number of glands in endometrium tissue of the rabbits in treatment group was increased significantly at the 1st and 2nd weeks（P<0.05），the percentages of the endometrial fibrosis areas were decreased significantly （P<0.05）， and the expression levels of integrin αVβ3 and LIF proteins in endometrium tissue were increased significantly （P<0.05）.In control group and treatment group， compared with the 1st week， there were no statistical differences in the number of glands，the percentages of endometrium fibrosis areas， and the expression levels of integrtin αVβ3 and LIF proteins at 2nd week（P>0.05）. But in model group， compared with the 1st week， the percentage of endometrium fibrosis area was decreased significantly at were 2nd week（P<0.01） and the expression levels of integrtin αVβ3 and LIF proteins in endometrium tissue were increased significantly （P<0.05）.

Conclusion

The placement of fresh amniotic membrane in the uterine cavity can reduce the degree of IUA and can improve the endometrial receptivity. The effect is fast and lasting.

Key words: uterine cavity adhesion, fresh amniotic membrane, integrin αVβ3, leukemia inhibitory factor, endometrial receptivity

中图分类号:

R711.74

范艳艳,王娜,徐文,葛亚杰,王丽娜. 新鲜羊膜对兔宫腔黏连的预防作用及对子宫内膜容受性的改善作用[J]. 吉林大学学报(医学版), 2021, 47(4): 990-998.

Yanyan FAN,Na WANG,Wen XU,Yajie GE,Lina WANG. Preventive effect of fresh amniotic membrane on intrauterine adhesion and its improvement effect on endometrial receptivity in rabbits[J]. Journal of Jilin University(Medicine Edition), 2021, 47(4): 990-998.

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参考文献 24

1	ZHU H Y， GE T X， PAN Y B， et al. Advanced role of hippo signaling in endometrial fibrosis［J］. Chin Med J， 2017， 130（22）： 2732-2737.
2	ABUDUKEYOUMU A， LI M Q， XIE F. Transforming growth factor-β1 in intrauterine adhesion［J］.Am J Reprod Immunol，2020，84（2）： e13262.
3	CHEN Y， LIU L， LUO Y， et al. Prevalence and impact of chronic endometritis in patients with intrauterine adhesions： a prospective cohort study［J］. J Minim Invasive Gynecol， 2017， 24（1）： 74-79.
4	CERVELLÓ I， CERVELLÓ I， GIL-SANCHIS C， et al. Human CD133（+） bone marrow-derived stem cells promote endometrial proliferation in a murine model of Asherman syndrome［J］. Fertil Steril， 2015， 104（6）： 1552-1560.
5	SANTAMARIA X， ISAACSON K， SIMÓN C. Asherman’s Syndrome： it may not be all our fault［J］. Hum Reprod， 2018， 33（8）： 1374-1380.
6	SONAN Y， AOKI S， ENOMOTO K， et al. Placenta accreta following hysteroscopic lysis of adhesions caused by Asherman’s syndrome： a case report and literature review［J］. Case Rep Obstet Gynecol， 2018， 2018： 1-4.
7	HOOKER A， FRAENK D， BRÖLMANN H， et al. Prevalence of intrauterine adhesions after termination of pregnancy： a systematic review［J］. Eur J Contracept Reprod Health Care， 2016， 21（4）： 329-335.
8	GILMAN A R， DEWAR K M， RHONE S A， et al. Intrauterine adhesions following miscarriage： look and learn［J］. J Obstet Gynaecol Can， 2016，38（5）： 453-457.
9	SHAKOURI-MOTLAGH A， KHANABDALI R， HEATH D E， et al. The application of decellularized human term fetal membranes in tissue engineering and regenerative medicine （TERM）［J］. Placenta， 2017， 59： 124-130.
10	GAN L， DUAN H， XU Q， et al. Human amniotic mesenchymal stromal cell transplantation improves endometrial regeneration in rodent models of intrauterine adhesions［J］. Cytotherapy， 2017， 19（5）： 603-616.
11	LI C， CAI A， SUN C， et al. The study on the safety and efficacy of amnion graft for preventing the recurrence of moderate to severe intrauterine adhesions［J］. Genes Dis， 2020， 7（2）： 266-271.
12	CHEN Y， FEI W D， ZHAO Y C， et al. Sustained delivery of 17β-estradiol by human amniotic extracellular matrix （HAECM） scaffold integrated with PLGA microspheres for endometrium regeneration［J］. Drug Deliv， 2020， 27（1）： 1165-1175.
13	DARRIBÈRE T， SKALSKI M， COUSIN H， et al. Integrins： Regulators of embryogenesis［J］. Biol Cell， 2000，92（1）：5-25.
14	MARWOOD M， VISSER K， SALAMONSEN L A， et al. Interleukin-11 and leukemia inhibitory factor regulate the adhesion of endometrial epithelial cells： implications in fertility regulation［J］. Endocrinology， 2009，150（6）：2915-2923.
15	LIU F， ZHU Z J， LI P， et al. Creation of a female rabbit model for intrauterine adhesions using mechanical and infectious injury［J］. J Surg Res，2013，183（1）： 296-303.
16	蔡慧华，何援利，李慧娟，等. 宫腔粘连大鼠模型的建立及其与白细胞介素21的相关性分析［J］. 医学研究生学报， 2015， 28（4）： 346-349.
17	CHI Y G， HE P， LEI L， et al. Transdermal estrogen gel and oral aspirin combination therapy improves fertility prognosis via the promotion of endometrial receptivity in moderate to severe intrauterine adhesion［J］. Mol Med Rep， 2018. DOI：10.3892/mmr.2018.8685 . doi: 10.3892/mmr.2018.8685
18	LIU L L， HUANG X W， XIA E L， et al. A cohort study comparing 4 mg and 10 mg daily doses of postoperative oestradiol therapy to prevent adhesion reformation after hysteroscopic adhesiolysis［J］. Hum Fertil， 2019， 22（3）： 191-197.
19	ZHU R， DUAN H， GAN L， et al. Comparison of intrauterine suitable balloon and Foley balloon in the prevention of adhesion after hysteroscopic adhesiolysis［J］. BioMed Res Int， 2018， 2018： 1-6.
20	PABUÇCU E G， KOVANCI E， ŞAHIN Ö， et al. New crosslinked hyaluronan gel， intrauterine device， or both for the prevention of intrauterine adhesions［J］. J Soc Laparoendosc Surg，2019.DOI：10.4293/jsls. 2018.00108 . doi: 10.4293/jsls. 2018.00108
21	秦琰，高金芳，王蔼明.宫腔粘连术后放置三角子宫球囊的临床疗效观察［J］. 解放军医学杂志，2019，44（8）： 717-720.
22	NAITO M， OGURA N， YAMANASHI T， et al. Prospective randomized controlled study on the validity and safety of an absorbable adhesion barrier （Interceed^®） made of oxidized regenerated cellulose for laparoscopic colorectal surgery［J］.Asian J Endosc Surg，2017，10（1）：7-11.
23	HOU X J， LIU Y， STREULI I， et al. Endometrial regeneration in Asherman’s syndrome： clinical and translational evidence of stem cell therapies［J］. Curr Stem Cell Res Ther， 2019， 14（6）： 454-459.
24	ORHUE A A， AZIKEN M E， IGBEFOH J O. A comparison of two adjunctive treatments for intrauterine adhesions following lysis［J］. Int J Gynaecol Obstet， 2003， 82（1）： 49-56.

Group	Number of glands
Group	1st week	2nd week
Control	8.720±0.885	8.750±0.750
Model	3.320±0.668^*	3.750±0.556^*
Teatment	7.200±0.525^*△	7.580±0.577^*△
P	<0.05	<0.05

Group	Percentage of endometrial fibrosis area
Group	1st week	2nd week
Control	16.61±2.75	16.57±2.65
Model	46.68±3.32^*	39.70±2.03^*#
Treatment	20.50±1.92^*△	19.97±3.42^*△
P	<0.01	<0.01

Group	Expression level of integrin αVβ3 protein
Group	1st week	2nd week
Control	5.11±0.18	5.12±0.18
Model	1.89±0.02^*	2.06±0.13^*#
Treatment	4.19±0.14^*△	4.37±0.23^*△
P	<0.01	<0.01

Group	Expression level of LIF protein
Group	1st week	2nd week
Control	5.16±0.24	5.08±0.13
Model	1.97±0.22^*	2.14±0.13^*#
Treatment	4.79±0.15^*△	4.22±0.20^*△
P	<0.05	<0.05

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[4]	姜秋，倪雪岩，聂代邦，孙宏晨. 小鼠胚胎成纤维细胞的转染与阳性细胞的初步筛选[J]. J4, 2008, 34(6): 990-993.
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[6]	李明堂，姜勇，付玉. 猪白血病抑制因子多克隆抗体的制备及其免疫定位[J]. J4, 2008, 34(1): 78-81.