Effect of Bushen Huoxue Decoction on endometrial morphology of thin endometrial rats treated with endometrial stem cell transplantation

Xiaodan YIN; Junqin HE; Mingwei XIN; Ying WU; Qian HAN; Wei YANG

doi:10.16305/j.1007-1334.2021.2004166

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Effect of Bushen Huoxue Decoction on endometrial morphology of thin endometrial rats treated with endometrial stem cell transplantation

更新时间：2022-08-03

- Effect of Bushen Huoxue Decoction on endometrial morphology of thin endometrial rats treated with endometrial stem cell transplantation
- Shanghai Journal of Traditional Chinese Medicine Vol. 55, Issue 7, Pages: 68-74,100(2021)
- 作者机构：
  
  1.首都医科大学附属北京妇产医院中医科（北京　100026）
- 作者简介：
- 基金信息：
- DOI：10.16305/j.1007-1334.2021.2004166
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Xiaodan YIN, Junqin HE, Mingwei XIN, et al. Effect of Bushen Huoxue Decoction on endometrial morphology of thin endometrial rats treated with endometrial stem cell transplantation. [J]. Shanghai Journal of Traditional Chinese Medicine 55(7):68-74,100(2021)
DOI：

Xiaodan YIN, Junqin HE, Mingwei XIN, et al. Effect of Bushen Huoxue Decoction on endometrial morphology of thin endometrial rats treated with endometrial stem cell transplantation. [J]. Shanghai Journal of Traditional Chinese Medicine 55(7):68-74,100(2021) DOI： 10.16305/j.1007-1334.2021.2004166.

摘要

目的,2,探讨补肾活血方对子宫内膜干细胞（EDSCs）移植治疗薄型子宫内膜模型大鼠子宫内膜形态学的影响。,方法,2,从8周龄雌性SD大鼠子宫内膜中分离提取EDSCs，采用机械分离与两步酶消法，实验选用纯度较高的第3代细胞，分别采用尾静脉（IV）和宫腔注射（IU）的方法移植到实验大鼠体内。采用宫腔灌注95%无水乙醇及灌胃羟基脲加尾静脉注射高分子右旋糖酐方法建立肾虚血瘀薄型子宫内膜联合模型。将实验大鼠分为正常组、模型组、戊酸雌二醇组、EDSCs+IV组、EDSCs+IU组、EDSCs+IV+中药组、EDSCs+IU+中药组，每组8只。各组大鼠给予相应的方法干预后，经过3个动情周期处死后取材。采用HE染色观察各组子宫内膜厚度、腺体和血管分布情况，免疫组化、Western blot检测角蛋白、波形蛋白的表达情况。,结果,2,在改善子宫内膜厚度方面，EDSCs+IU组、EDSCs+IV+中药组及EDSCs+IU+中药组较模型组内膜厚度均增加，差异有统计学意义（,P,<,0.01）；在增加子宫内膜腺体数方面，EDSCs+IV组较模型组腺体数增加，差异有统计学意义（,P,<,0.05），EDSCs+IU组、EDSCs+IV+中药组、EDSCs+IU+中药组较模型组腺体数增加，差异有统计学意义（,P,<,0.01）；在增加子宫内膜血管数方面，EDSCs+IV组、EDSCs+IU组、EDSCs+IV+中药组、EDSCs+IU+中药组较模型组血管数增加，差异有统计学意义（,P,<,0.01）；在改善角蛋白表达方面，EDSCs+IV组、EDSCs+IU组、EDSCs+IV+中药组、EDSCs+IU+中药组较模型组表达量升高，差异有统计学意义（,P,<,0.01）；在改善波形蛋白表达方面，EDSCs+IV组、EDSCs+IU组、EDSCs+IV+中药组、EDSCs+IU+中药组较模型组表达量升高，差异有统计学意义（,P,<,0.01）；在改善子宫内膜厚度、免疫组化检测波形蛋白表达方面宫腔注射EDSCs与尾静脉注射EDSCs比较，差异有统计学意义（,P,<,0.01）；EDSCs+IV+中药组与EDSCs+IV组比较、EDSCs+IU+中药组与EDSCs+IU组比较子宫内膜厚度显著增加，差异有统计学意义（,P,<,0.01）。,结论,2,宫腔注射EDSCs在增加子宫内膜厚度、免疫组化检测波形蛋白表达方面更优于尾静脉注射EDSCs；加用补肾活血方后有助于增加子宫内膜厚度。

Abstract

Objective,2,To investigate the effect of Bushen Huoxue Decoction on endometrial morphology in rats with thin endometrium treated by transplantation of endometrial stem cells (EDSCs).,Methods,2,EDSCs were isolated from the endometrium of 8-week-old female SD rats by the methods of mechanical separation and two-step enzyme elimination. The third generation cells with high purity were selected and transplanted into the experimental rats by tail vein and intrauterine injection respectively. The combined model of thin endometrium with kidney deficiency and blood stasis was established by infusing 95% ethanol without water into the uterine cavity and injecting hydroxyurea and high molecular dextran into the tail vein of female SD rats. The experimental rats were divided into normal control group, model group, estradiol valerate group, EDSCs + IV group, EDSCs + IU group, EDSCs + IV+ traditional Chinese medicine (TCM) group and EDSCs+IU+TCM group, with 8 rats in each group. After intervention with corresponding methods, the rats in each group were killed after 3 estrous cycles. HE staining was used to observe the thickness of uterus, the number of glands and blood vessels, and the expressions of keratin and vimentin by immunohistochemistry and Western blot.,Results,2,In terms of improving endometrial thickness, the endometrial thickness in the EDSCs+IU group, the EDSCs+IV+TCM group and the EDSCs+IU+TCM group was significantly higher than that in the model group(,P,<,0.01); in terms of increasing the number of endometrial glands, the number of glands in the EDSCs+IV group was significantly higher than that in the model group (,P,<,0.05), and that in the EDSCs+IU group, the EDSCs+IV+TCM group and the EDSCs+IU+ TCM group was significantly higher than that in the model group (,P,<,0.01); in terms of increasing the number of endometrial blood vessels, the number of blood vessels in the EDSCs+IV group, the EDSCs+IU group, the EDSCs+IV+TCM group and the EDSCs+IU+TCM group was significantly higher than that in the model group (,P,<,0.01); in terms of improving the expression of keratin, the expression of keratin in the EDSCs+IV group, the EDSCs+IU group, the EDSCs+IV+TCM group and the EDSCs+IU+TCM group was significantly higher than that in the model group (,P,<,0.01); in terms of improving the expression of vimentin, the expression of vimentin in the EDSCs+IV group, the EDSCs+IU group, the EDSCs+IV+TCM group and the EDSCs+IU+ TCM group was significantly higher than that in the model group (,P,<,0.01). There was significant difference between intrauterine injection of EDSCs and tail vein injection of EDSCs in improving the thickness of endometrium and the expression of vimentin detected by immunohistochemistry (,P,<,0.01). The endometrial thickness in the EDSCs+IV+TCM group was significantly higher than that in the EDSCs+IV group, and that in the EDSCs+IU+TCM group was significantly higher than that in the EDSCs+IU group (,P,<,0.01).,Conclusion,2,Intrauterine injection of EDSCs is better than tail vein injection in increasing endometrial thickness and vimentin expression detected by immunohistochemistryl, and adding Bushen Huoxue Decoction helps to increase the thickness of endometrium.

关键词

补肾活血方薄型子宫内膜模型大鼠子宫内膜干细胞移植子宫内膜形态学中药研究

Keywords

Bushen Huoxue Decoctionthin endometriummodel ratsendometrial stem cell transplantationendometrial morphologytraditional Chinese herbol medicine research

references

AZIZI R, AGHEBATI-MALEKI L, NOURI M, et al. Stem cell therapy in asherman syndrome and thin endometrium: stem cell- based therapy [J]. Biomed Pharmacother, 2018(102): 333-343.

KIM H, SHIN J E, KOO H S, et al. Effect of autologous platelet-rich plasma treatment on refractory thin endometrium during the frozen embryo transfer cycle: a pilot study[J]. Front Endocrinol (Lausanne), 2019(10): 61.

CASPER R F. It’s time to pay attention to the endometrium [J]. Fertil Steril, 2011, 96(3): 519-521.

DAVAR R, DEHGHANI FIROUZABADI R, CHAMAN ARA K. Dilatation and curettage effect on the endometrial thickness[J]. Iran Red Crescent Med J, 2013, 15(4): 350-355.

TALUKDAR N, BENTOV Y, CHANG P T, et al. Effect of long-term combined oral contraceptive pill use on endometrial thickness [J]. Obstet Gynecol, 2012, 120(2Pt1):348-354.

王利公，杨新园，王伟.子宫内膜基质干细胞的分离培养研究[J].浙江大学学报（医学版），2013, 42(3): 311-318.

GARGETT C E, NGUYEN H P, YE L. Endometrial regeneration and endometrial stem/progenitor cells [J]. Rev Endocr Metab Disord, 2012, 1(4):235-251.

高红，赵静，李艳萍. 大鼠薄型子宫内膜模型的建立和鉴定[J]. 生命科学研究，2011, 15(5): 426-431.

林继红，杨海英. 雌性大鼠肾虚模型初探[J]. 中国比较医学杂志，2010, 20(10): 78.

宋程程，王志斌，苏斌. 常用大鼠血瘀证模型的比较研究[J]. 北京中医药大学学报，2014, 37 (2): 94-98.

尹晓丹，何军琴，王景尚，等. 补肾活血方对肾虚血瘀薄型子宫内膜大鼠子宫内膜容受性的影响研究[J]. 中国全科医学，2018, 21(6): 683-688.

魏伟，吴希美，李元建. 药理实验方法学[M]. 北京：人民卫生出版，2010: 76-180.

GARGETT C E, MASUDA H. Adult stem cells in the endometrium[J].Mol Hum Reprod, 2010, 16(11): 818-834.

PRIANISHNIKOV V A.On the concept of stem cell and a model of functional morphological stracture of the endometrium[J]. Conception, 1978, 18(3): 213-223.

TAYLOR H S. Endometrial cells derived from donor stem cells in bone marrow transplant recipients[J]. JAMA, 2004, 292(1): 81-85.

尹晓丹，杨维，辛明蔚，等. 补肾活血方对薄型子宫内膜排卵障碍患者子宫内膜容受性及临床妊娠率的影响[J]. 上海中医药杂志，2018, 52(3): 55-57.

XIN M W, HE J Q, YANG W, et al. Wenshen Yangxue decoction improves endometrial receptivity recovery and promotes endometrial angiogenesis in a rat model [J]. Pharm Biol, 2018, 56(1): 573-579.

尹晓丹，何军琴，杨维. 补肾活血方对薄型子宫内膜干细胞Wnt/β-catenin信号通路的影响[J]. 北京中医药大学学报，2018, 41(2): 118-123.

ZUO W, XIE B, LI C, et al. The clinical application of endometrial mesenchymal stem cells[J]. Biopreserv Biobank, 2018, 16(2): 158-164.

MENG X, ICHIM T E, ZHANG J, et al. Endometrial regenerative cells: a novel stem cell population[J]. J Transl Med, 2007(5): 57.

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