补肾活血方对子宫内膜干细胞移植治疗薄型子宫内膜大鼠子宫内膜CD34、整合素α_(ν)β_(3)、LIF表达的影响  

Effect of Bushen Huoxue Recipe on Expression of CD34,Integrin α_(ν)β_(3) and LIF in Endometrium of Rats with Endometrial Stem Cells Transplantation Treatment of Thin Endometrium

作　　者：尹晓丹[1] 杨维[1] 辛明蔚[1] 武颖[1] 何军琴[1] YIN Xiaodan;YANG Wei;XIN Mingwei;WU Ying;HE Junqin(Department of Traditional Chinese Medicine,Beijing Obstetrics and Gynecology Hospital,Capital Medical University.Beijing Maternal and Child Health Care Hospital,Beijing 100026,China)

机构地区：[1]首都医科大学附属北京妇产医院/北京妇幼保健院中医科,北京100026

出　　处：《医学综述》2022年第5期997-1003,共7页Medical Recapitulate

基　　金：国家自然科学基金(82004134);北京中医药科技发展资金项目(QN2018-04)。

摘　　要：目的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CD34、整合素α_(ν)β_(3)、白细胞抑制因子(LIF)表达的影响。方法从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化法,实验选用纯度较高的第3代细胞,分别采用尾静脉和宫腔注射的方法移植到实验大鼠体内;采用雌性SD大鼠宫腔灌注95%无水乙醇及灌胃羟基脲+尾静脉注射高分子右旋糖苷方法建立肾虚血瘀型薄型子宫内膜联合模型;将42只实验大鼠依据随机数字法分为正常对照组、模型组、戊酸雌二醇组、EDSCs+尾静脉注射组、EDSCs+宫腔注射组、EDSCs+尾静脉注射+中药组、EDSCs+宫腔注射+中药组,每组6只;各干预组分别给予BMSCs静脉或宫腔移植并联合中药干预后经过大鼠3个动情周期,处死取材。采用免疫组织化学、Western blot检测子宫内膜中CD34、整合素α_(ν)β_(3)及LIF的表达情况。结果免疫组织化学法检测结果显示,模型组大鼠子宫内膜CD34、整合素α_(ν)β_(3)、LIF的表达低于正常对照组(0.080±0.008比0.210±0.015、0.088±0.018比0.267±0.037、0.065±0.011比0.239±0.019,均P<0.01),EDSCs+宫腔注射+中药组大鼠子宫内膜整合素α_(ν)β_(3)、LIF的表达高于EDSCs+宫腔注射组(P<0.05)。Western blot检测结果显示,模型组大鼠子宫内膜CD34、整合素α_(ν)β_(3)、LIF的表达低于正常对照组(P<0.01),EDSCs+宫腔注射组大鼠子宫内膜CD34表达高于EDSCs+尾静脉注射组(P<0.05),EDSCs+尾静脉注射+中药组、EDSCs+宫腔注射+中药组高于EDSCs+宫腔注射组(P<0.05);EDSCs+宫腔注射组、EDSCs+尾静脉注射+中药组整合素α_(ν)β_(3)的表达高于EDSCs+尾静脉注射组(P<0.05),EDSCs+宫腔注射+中药组高于EDSCs+宫腔注射组(P<0.05)。结论补肾活血方能显著增加EDSCs移植治疗薄型子宫内膜大鼠子宫内膜CD34、整合素α_(ν)β_(3)、LIF的表达。Objective To investigate the effect of Bushen huoxue recipe on the endometrial expression of CD34,integrinα_(ν)β_(3) and leukocyte inhibitory factor(LIF)in the thin endometrial model rats treated by endometrial stem cells(EDSCs)transplantation.Methods EDSCs were isolated and extracted from the endometrium of 8-week-old female SD rats by mechanical separation and two-step enzymatic digestion.The third generation cells with higher purity were selected in the experiment and transplanted by tail vein and intrauterine injection respectively.In the experimental rats,female SD rats were perfused with 95%absolute ethanol and hydroxyurea in the uterine cavity and injected with polymer dextran into the caudal vein to establish a combined model of kidney-deficiency and blood stasis type thin endometrium.42 experimental rats were divided into a normal control group,a model group,an estradiol valerate group,an EDSCs+caudal vein injection group,an EDSCs+intrauterine injection group,an EDSCs+caudal vein injection+traditional Chinese medicine(TCM)group,an EDSCs+intrauterine injection+TCM group,6 rats in each group.Each intervention group received intravenous or intrauterine transplantation of BMSCs and combined with Traditional Chinese medicine intervention,and after three estrous cycles,they were killed.Immunohistochemistry and Western blot were used to detect the expression of vascular endothelial cell marker protein CD34,integrinα_(ν)β_(3) and LIF protein.Results Immunohistochemical analysis showed that the expression of CD34,integrinα_(ν)β_(3) and LIF in endometrium of rats in the model group was lower than those in the control group(0.080±0.008 vs 0.210±0.015,0.088±0.018 vs 0.267±0.037,0.065±0.011 vs 0.239±0.019,all P<0.01),and the expression of integrinα_(ν)β_(3) and LIF in the EDSCs+intrauterine injection+TCM group was higher than those in the EDSCs+intrauterine injection group(P<0.05).The Western blot analysis showed that the expression of CD34,integrinα_(ν)β_(3) and LIF in endometrium of rats in the

关 键 词：薄型子宫内膜 补肾活血方 子宫内膜干细胞 CD34 整合素α_(ν)β_(3) 白细胞抑制因子 

分 类 号：R711.76[医药卫生—妇产科学]