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1.南京中医药大学针灸推拿·养生康复学院,南京210046
2.徐州医科大学,江苏徐州221004
Received:18 August 2022,
Revised:05 December 2022,
Published:25 September 2023
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郝赛楠,夏良君,席瑾等.电针联合骨髓间充质干细胞移植通过调节SDF-1/CXCR4轴促进薄型子宫内膜修复的机制研究[J].针刺研究,2023,48(09):870-880.
HAO Sai-nan,XIA Liang-jun,XI Jin,et al.Electroacupuncture combined with bone marrow mesenchymal stem cell transplantation promotes repair of thin endometrium by regulating SDF-1/CXCR4 signaling[J].Acupuncture Research,2023,48(09):870-880.
郝赛楠,夏良君,席瑾等.电针联合骨髓间充质干细胞移植通过调节SDF-1/CXCR4轴促进薄型子宫内膜修复的机制研究[J].针刺研究,2023,48(09):870-880. DOI: 10.13702/j.1000-0607.20220942.
HAO Sai-nan,XIA Liang-jun,XI Jin,et al.Electroacupuncture combined with bone marrow mesenchymal stem cell transplantation promotes repair of thin endometrium by regulating SDF-1/CXCR4 signaling[J].Acupuncture Research,2023,48(09):870-880. DOI: 10.13702/j.1000-0607.20220942.
目的
2
观察电针联合骨髓间充质干细胞(BMSCs)移植对薄型子宫内膜形态、子宫内膜组织基质细胞衍生因子-1(SDF-1)/CXC趋化因子受体4(CXCR4)轴相关蛋白及mRNA的影响,探讨其修复薄型子宫内膜的机制。
方法
2
将健康SPF级性成熟雌性SD大鼠按随机数字表法分为空白组、模型组、BMSCs组、BMSCs+AMD3100组、BMSCs+电针组(联合组)、联合+AMD3100组,每组5只。除空白组外,其余5组采用95%乙醇宫腔注射的方法于大鼠动情期制备薄型子宫内膜模型。BMSCs组于造模第1、3、7天给予尾静脉注射BMSCs。BMSCs+AMD3100组在BMSCs组的基础上,以5 mg/kg的剂量给予腹腔注射AMD3100,每日1次;联合组在BMSCs组的基础上取“子宫”“关元”“三阴交”,每日给予电针刺激15 min;联合+AMD3100组在联合组干预基础上,以5 mg/kg的剂量给予腹腔注射AMD3100,每日1次。以上3组均干预3个动情周期。通过HE染色法观察大鼠子宫内膜形态,免疫组织化学法检测子宫内膜组织波形蛋白、角蛋白19的表达,Western blot法检测子宫内膜组织同源框(HOXA)10、白血病抑制因子(LIF)、SDF-1和CXCR4的蛋白表达水平,实时荧光定量PCR法检测大鼠子宫内膜组织SDF-1和CXCR4 mRNA的表达。
结果
2
与空白组比较,模型组大鼠子宫内膜腺体数目及子宫内膜组织波形蛋白、角蛋白19的阳性表达均减少(
P
<
0.01),HOXA10、LIF、CXCR4蛋白及CXCR4 mRNA表达水平均降低(
P
<
0.01),SDF-1蛋白及mRNA表达水平均升高(
P
<
0.05)。与模型组比较,BMSCs组和联合组大鼠子宫内膜腺体数目及子宫内膜组织中波形蛋白、角蛋白19的阳性表达增多(
P
<
0.05,
P
<
0.01),HOXA10、LIF、CXCR4蛋白及CXCR4 mRNA表达水平均升高(
P
<
0.01,
P
<
0.05);联合组SDF-1蛋白及mRNA表达水平升高(
P
<
0.05)。与BMSCs组比较,联合组子宫内膜腺体数目增多(
P
<
0.01),LIF、CXCR4蛋白及CXCR4 mRNA表达水平均升高(
P
<
0.01,
P
<
0.05);BMSCs+AMD3100组子宫内膜腺体数目及子宫内膜组织中波形蛋白、角蛋白19的阳性表达减少(
P
<
0.01),HOXA10、LIF、CXCR4蛋白及mRNA表达水平降低(
P
<
0.01)。与联合组比较,联合+AMD3100组大鼠子宫内膜腺体数目及子宫内膜组织波形蛋白、角蛋白19阳性表达均减少(
P
<
0.01),HOXA10、LIF、CXCR4蛋白及CXCR4 mRNA表达水平均降低(
P
<
0.01)。
结论
2
电针可增强与干细胞归巢相关的SDF-1和CXCR4信号分子的表达,促进移植的BMSCs向受损部位转移,从而促进薄型子宫内膜再生,修复子宫内膜损伤,改善子宫内膜容受性。
Objective
2
It is to explore, based on stromal cell derived factor 1 (SDF-1)/CXC chemokine receptor 4 (CXCR4) signal axis, whether the electroacupuncture (EA) combined with bone marrow mesenchymal stem cells (BMSCs) transplantation can promote thin endometrium regeneration and improve endometrial receptivity, so as to further study its mechanisms underlying improvement of promoting BMSCs homing to repair thin endometrium.
Methods
2
Thirty matured female SD rats were randomly divided into normal control , model , BMSCs transplantation (BMSCs), BMSCs+AMD3100 (a specific antagonist of CXCR4, BMSCs+AMD3100), BMSCs+EA, and BMSCs+EA+AMD3100 groups, with 5 rats in each group. The thin endometrial model was established by intrauterine injection of 95% ethanol during the period of estrus. Rats of the model group received intravenous injection of PBS solution (tail vein) on day 1, 3 and 7 of modeling and intraperitoneal injection of normal saline once daily for 3 estrous cycles. Rats of the BMSCs group received intravenous injection of BMSCs suspension on day 1,3 and 7 of modeling, and those of the BMSCs+EA group received BMSCs transplantation and EA stimulation. EA (2 Hz/15 Hz, 1 mA) was applied to “Guanyuan” (CV4) and bilateral “Sanyinjiao”(SP9), “Zigong” (EX-CA1) for 15 min, once daily for 3 estrous cycles. Rats of the BMSCs+AMD3100 group received intravenous injection of BMSCs suspension (1×10
6
/mL) and intraperitoneal injection of AMD3100 (5 mg/kg), and those of the BMSCs+EA+AMD3100 group received administration of BMSCs, AMD3100 and EA, with both groups being once daily for 3 estrous cycles. H.E. staining was used to observe histopathological changes of endometrium tissues, and immunohistochemistry was used to detect the expressions of cytokeratin (CK19) and vimentin in endometrium (for evaluating the damage and repair of endometrium). The expression levels of homeobox A10 (HOXA10), leukemia inhibitory factor (LIF), SDF-1 and CXCR4 proteins were detected by Western blot, and those of SDF-1 and CXCR4 mRNAs in the endometrium detected by real-time PCR.
Results
2
In comparison with the normal control group, the number of endometrial glands, the immunoactivity of CK19 and vimentin, the expression leve-ls of HOXA10, LIF and CXCR4 proteins and CXCR4 mRNA were significantly down-regulated (
P
<
0.01), and the expression levels of SDF-1 protein and mRNA significantly up-regulated (
P
<
0.05) in the model group. Compared with the model group, the number of endometrial glands, the immunoactivity of CK19 and vimentin, and the expression levels of HOXA10, LIF, CXCR4 proteins and CXCR4 mRNA in the BMSCs group, and the number of endometrial glands, the immunoactivity of CK19 and vimentin, the expression levels of HOXA10, LIF, CXCR4 proteins and CXCR4 mRNA, and SDF-1 protein and mRNA in the BMSCs+EA group were significantly up-regulated (
P
<
0.05,
P
<
0.01). Compared to the BMSCs group, the number of endometrial glands, and the expression levels of LIF, CXCR4 proteins and CXCR4 mRNA in the BMSCs+EA group were up-regulated (
P<
0.01,
P
<
0.05); the number of endometrial glands, the immunoactivity of CK19 and vimentin, the expression levels of HOXA10, LIF, CXCR4 proteins and CXCR4 mRNA in the BMSCs+AMD3100 group were down-regulated (
P
<
0.01). Compared to the BMSCs+EA group, the number of endometrial glands, the immunoactivity of CK19 and vimentin, the expression levels of HOXA10, LIF, CXCR4 proteins and CXCR4 mRNA in the BMSCs+EA+AMD3100 group were down-regulated (
P
<
0.01). Results of H.E. staining showed thin endometrium with absence of epithelial cells, and sparse glands and blood vessels, with smaller glandular cavity in the model group, which was relative milder in BMSCs and BMSCs+EA groups.
Conclusion
2
EA can promote the transfer of transplanted BMSCs to the damaged site through SDF-1/CXCR4 signaling related stem cell homing, thereby promoting thin endometrial regeneration, repairing endometrial injury, and improving endometrial tolerance in rats with thin endometrium.
SHUFARO Y , SIMON A , LAUFER N , et al . Thin unresponsive endometrium:a possible complication of surgical curettage compromising ART outcome [J]. J Assist Reprod Genet , 2008 , 25 ( 8 ): 421 - 425 .
MAHAJAN N , SHARMA S . The endometrium in assisted reproductive technology: how thin is thin? [J]. J Hum Reprod Sci , 2016 , 9 ( 1 ): 3 - 8 .
赵小萱 , 陈璐 , 姜月蓬 , 等 . 薄型子宫内膜的中西医研究进展 [J]. 湖南中医药大学学报 , 2017 , 37 ( 12 ): 1425 - 1430 .
ZHAO X X , CHEN L , JIANG Y P , et al . Advances in Chinese and western medicine of thin endometrium (in Chinese) [J]. Journal of Hunan University of Chinese Medicine , 2017 , 37 ( 12 ): 1425 - 1430 .
陈滢如 , 房繄恭 , 王飞 , 等 . 针刺辅助生殖功能的国外研究进展 [J]. 针刺研究 , 2014 , 39 ( 4 ): 329 - 332 .
CHEN Y R , FANG Y G , WANG F , et al . Development of researches on acupuncture-assisted fertilization outside of China (in Chinese) [J]. Acupuncture Research , 2014 , 39 ( 4 ): 329 - 332 .
XU X A , ZHU F Q , ZHANG M , et al . Stromal cell-derived factor-1 enhances wound healing through recruiting bone marrow-derived mesenchymal stem cells to the wound area and promoting neovascularization [J]. Cells Tissues Organs , 2013 , 197 ( 2 ): 103 - 113 .
LIU H C , YANG K Y , XIN T , et al . BMSCs [J]. Artif Cells Blood Substit Biotechnol , 2012 , 40 ( 5 ): 331 - 337 .
SALAZAR T E , RICHARDSON M R , BELI E , et al . Electroacupuncture promotes central nervous system-dependent release of mesenchymal stem cells [J]. Stem Cells , 2017 , 35 ( 5 ): 1303 - 1315 .
王玉影 , 曹奕 , 陈倩倩 , 等 . 针灸通督法对缺血性脑卒中患者BMSCs的影响 [J]. 上海针灸杂志 , 2015 , 34 ( 2 ): 101 - 104 .
WANG Y Y , CAO Y , CHEN Q Q , et al . Acupuncture-moxibustion with Tong du method on the bone marrow stem cells in patients with ischemic stroke (in Chinese) [J]. Shanghai Journal of Acupuncture and Moxibustion , 2015 , 34 ( 2 ): 101 - 104 .
唐一鸣 , 徐慧军 . 中西医结合治疗薄型子宫内膜合并不孕症验案浅析 [J]. 中国民族民间医药 , 2018 , 27 ( 17 ): 83 - 85 .
TANG Y M , XU H J . Clinical analysis of combination of traditional Chinese medicine and western medicine in treatment of thin endometrium with infertility (in Chinese) [J]. Chinese Journal of Ethnomedicine and Ethnopharmacy , 2018 , 27 ( 17 ): 83 - 85 .
艾丹 , 钟丽珍 , 王丹 , 等 . 中药配合针灸理疗在宫腔粘连术后子宫内膜修复治疗中的应用研究 [J]. 医学理论与实践 , 2021 , 34 ( 16 ): 2828 - 2830 .
AI D , ZHONG L Z , WANG D , et al . Study on the application of traditional Chinese medicine combined with acupuncture and physiotherapy in endometrial repair after intrauterine adhesions (in Chinese) [J]. The Journal of Medical Theory and Practice , 2021 , 34 ( 16 ): 2828 - 2830 .
张莹 . 中药、针灸联合腔内理疗改善薄型子宫内膜的临床疗效观察 [J]. 反射疗法与康复医学 , 2022 , 3 ( 4 ): 11 - 14 .
ZHANG Y . Clinical observation on improving thin endometrium with traditional Chinese medicine, acupuncture and endovascular physiotherapy (in Chinese) [J]. Reflexology and Rehabilitation Medicine , 2022 , 3 ( 4 ): 11 - 14 .
XIA L J , MENG Q Y , XI J , et al . The synergistic effect of electroacupuncture and bone mesenchymal stem cell transplantation on repairing thin endometrial injury in rats [J]. Stem Cell Res Ther , 2019 , 10 : 244 .
XI J , CHENG J E , JIN C C , et al . Electroacupuncture improves pregnancy outcomes in rats with thin endometrium by promoting the expression of pinopode-related molecules [J]. BioMed Res Int , 2021 , 2021 : 1 - 12 .
孟庆宇 , 席瑾 , 夏良君 , 等 . 电针联合骨髓间充质干细胞注射对大鼠薄型子宫内膜雌、孕激素受体的影响 [J]. 针刺研究 , 2021 , 46 ( 5 ): 385 - 390 .
MENG Q Y , XI J , XIA L J , et al . Effect of combined admini-stration of electroacupuncture and mesenchymal stem cells on expression of endometrium estrogen receptor and progesterone receptor in thin endometrium rats (in Chinese) [J]. Acupuncture Research , 2021 , 46 ( 5 ): 385 - 390 .
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 .
GAO H , ZHAO J , LI Y P . Establishment and identification of rat thin endometrium model (in Chinese) [J]. Life Science Research , 2011 , 15 ( 5 ): 426 - 431 .
赵静 . 骨髓间充质干细胞移植治疗薄型子宫内膜的实验研究 [D]. 长沙 : 中南大学 , 2013 .
ZHAO J . The experimental study of therapeutic effect of bone marrow mesenchymal stem cells transplantation on thin endometrium (in Chinese) [D]. Changsha : Central South University , 2013 .
中国针灸学会 . 实验动物常用穴位名称与定位第2部分:大鼠 [J]. 针刺研究 , 2021 , 46 ( 4 ): 351 - 352 .
China Association of Acupuncture-Moxibustion . Nomenclature and location of acupoints commonly used in experimental animals. Part 2: rats (in Chinese) [J]. Acupuncture Research , 2021 , 46 ( 4 ): 351 - 352 .
张莹 , 曹圣 , 文绍敦 , 等 . 针刺对多囊卵巢综合征胰岛素抵抗大鼠生殖内分泌及磷酸化蛋白激酶B蛋白表达的影响 [J]. 针刺研究 , 2018 , 43 ( 10 ): 622 - 626 .
ZHANG Y , CAO S , WEN S D , et al . Effect of acupuncture on reproductive endocrine and p-Akt protein expression in polycystic ovary syndrome rats with insulin resistance (in Chinese) [J]. Acupuncture Research , 2018 , 43 ( 10 ): 622 - 626 .
王聪聪 , 刘睿 , 袁淑绘 , 等 . CXCLl2在大鼠短暂性局灶性脑缺血损伤超早期中的作用 [J]. 中风与神经疾病杂志 , 2015 , 32 ( 12 ): 1095 - 1098 .
WANG C C , LIU R , YUAN S H , et al . The role of CXCL12 on the ultra-early time of transient focal brain ischemic injury in rats (in Chinese) [J]. Journal of Apoplexy and Nervous Diseases , 2015 , 32 ( 12 ): 1095 - 1098 .
黄雪花 , 刘宁洁 , 戴丽华 , 等 . AMD3100对坐骨神经分支选择性损伤模型大鼠的影响 [J]. 上海交通大学学报(医学版) , 2016 , 36 ( 1 ): 13 - 16, 22 .
HUANG X H , LIU N J , DAI L H , et al . Effects of AMD3100 on rats of spared nerve injury model (in Chinese) [J]. Journal of Shanghai Jiao Tong University(Medical Science) , 2016 , 36 ( 1 ): 13 - 16, 22 .
秦爱华 , 林燕 , 邓佳妮 . 补肾调经汤联合温和灸法对辅助生殖子宫内膜薄的临床观察 [J]. 世界中医药 , 2019 , 14 ( 12 ): 3364 - 3368 .
QIN A H , LIN Y , DENG J N . Clinical observation on Bushen Tiaojing Decoction combined with mild moxibustion for assisted reproductive thin endometrium patients (in Chinese) [J]. World Chinese Medicine , 2019 , 14 ( 12 ): 3364 - 3368 .
GARCIA-VELASCO J A , ACEVEDO B , ALVAREZ C , et al . Strategies to manage refractory endometrium: state of the art in 2016 [J]. Reprod Biomed Online , 2016 , 32 ( 5 ): 474 - 489 .
RANISAVLJEVIC N , RAAD J , ANAHORY T , et al . Embryo transfer strategy and therapeutic options in infertile patients with thin endometrium: a systematic review [J]. J Assist Reprod Genet , 2019 , 36 ( 11 ): 2217 - 2231 .
李乔乔 , 吴振强 , 张丽君 . 骨髓间充质干细胞的定向分化潜能 [J]. 中国组织工程研究 , 2017 , 21 ( 25 ): 4082 - 4087 .
LI Q Q , WU Z Q , ZHANG L J . Directional differentiation of bone marrow mesenchymal stem cells (in Chinese) [J]. Chinese Journal of Tissue Engineering Research , 2017 , 21 ( 25 ): 4082 - 4087 .
LIU H B , XUE W J , GE G Q , et al . Hypoxic preconditio-ning advances CXCR4 and CXCR7 expression by activating HIF-1α in MSCs [J]. Biochem Biophys Res Commun , 2010 , 401 ( 4 ): 509 - 515 .
钱洁 , 杜鑫 . 针灸在辅助生殖技术中的应用效果 [J]. 中国全科医学 , 2016 , 19 ( S1 ): 425 - 427 .
QIAN J , DU X . Application effect of acupuncture and moxibustion in assisted reproductive technology (in Chinese) [J]. Chinese General Practice , 2016 , 19 ( S1 ): 425 - 427 .
YANG H , LI J , ZHOU Y C , et al . Research progress on acupuncture-moxibustion in treatment of refractory infertility [J]. World J Acu-Moxibust , 2020 , 30 ( 2 ): 125 - 129 .
汪秀梅 , 袁红丽 , 刘欢 , 等 . 针灸对不孕症薄型子宫内膜临床疗效观察 [C]//中国针灸学会. 2017世界针灸学术大会暨2017中国针灸学会年会论文集 . 北京 , 2017 .
WANG X M , YUAN H L , LIU H , et al . Clinical observation on the effect of acupuncture on thin endometrium of infertile women (in Chinese) [C]// China Association of Acupuncture-Moxibustion . Proceedings of 2017 World Congress of Acupuncture and Moxibustion and 2017 Annual Conference of Chinese Acupuncture and Moxibustion Society . Beijing , 2017 .
陈茜 , 梁凤霞 , 吴松 , 等 . 标本配穴的理论探讨与临床应用 [J]. 中国针灸 , 2018 , 38 ( 5 ): 505 - 509 .
CHEN Q , LIANG F X , WU S , et al . Theoretic exploration and clinical application of acupoint combination based on Biaoben theory (in Chinese) [J]. Chinese Acupuncture & Moxibustion , 2018 , 38 ( 5 ): 505 - 509 .
程贤鹦 , 曾淑梅 , 李淑慧 , 等 . 关元穴穴位按摩联合中药封包对镇痛分娩后膀胱排尿功能恢复和尿潴留预防作用研究 [J]. 中国医药导报 , 2020 , 17 ( 22 ): 160 - 162, 170 .
CHENG X Y , ZENG S M , LI S H , et al . Study on the preventive effect of Guanyuan acupoint massage combined with traditional Chinese medicine package therapy on the recovery of bladder micturition function and the prevention of urinary retention in parturients with analgesia delivery (in Chinese) [J]. China Medical Herald , 2020 , 17 ( 22 ): 160 - 162, 170 .
高伟娜 , 唐潇 , 呙月 , 等 . 针刺对胚胎着床障碍大鼠子宫内膜CCR2的影响 [J]. 中西医结合研究 , 2013 , 5 ( 4 ): 176 - 180 .
GAO W N , TANG X , GUO Y , et al . The effect of acupuncture on CCR2in endometrium of rat with embryo implantation dysfunction (in Chinese) [J]. Research of Integrated Traditional Chinese and Western Medicine , 2013 , 5 ( 4 ): 176 - 180 .
张汝月 , 李瑞娇 , 张引红 , 等 . 薄型子宫内膜小鼠模型的免疫组化鉴定与生育力评估 [J]. 中国比较医学杂志 , 2018 , 28 ( 12 ): 19 - 26 .
ZHANG R Y , LI R J , ZHANG Y H , et al . Immunohistochemical identification and fertility assessment of a mouse model of thin endometrium (in Chinese) [J]. Chinese Journal of Comparative Medicine , 2018 , 28 ( 12 ): 19 - 26 .
何东杰 , 卫爱武 , 许丽绵 , 等 . 益肾活血丸调控Wnt7a/HOXA10改善内膜容受性的机理研究 [J]. 中医学报 , 2022 , 37 ( 6 ): 1261 - 1268 .
HE D J , WEI A W , XU L M , et al . Mechanism of Yishen Huoxue pills in regulating Wnt7a/HOXA10 and improving endometrial receptivity (in Chinese) [J]. Acta Chinese Medicine , 2022 , 37 ( 6 ): 1261 - 1268 .
TAPIA A , SALAMONSEN L A , MANUELPILLAI U , et al . Leukemia inhibitory factor promotes human first trimester extravillous trophoblast adhesion to extracellular matrix and secretion of tissue inhibitor of metalloproteinases-1 and-2 [J]. Hum Reprod , 2008 , 23 ( 8 ): 1724 - 1732 .
朱磊 , 连紫宇 , 胡静怡 , 等 . 健脾补肾、清肠化湿复方及其拆方对骨髓间充质干细胞增殖趋化的影响及机制研究 [J]. 中华中医药杂志 , 2021 , 36 ( 8 ): 4568 - 4572 .
ZHU L , LIAN Z Y , HU J Y , et al . Study on the effects and mechanism of Jianpi Bushen, Qingchang Huashi Compound and its decomposed prescriptions on proliferation and chemotaxis of BMSCs (in Chinese) [J]. China Journal of Traditional Chinese Medicine and Pharmacy , 2021 , 36 ( 8 ): 4568 - 4572 .
CUN Y P , DIAO B , ZHANG Z M , et al . Role of the stromal cell derived factor-1 in the biological functions of endothelial progenitor cells and its underlying mechanisms [J]. Exp Ther Med , 2021 , 21 ( 1 ): 39 .
KAWAGUCHI A , ORBA Y , KIMURA T , et al . Inhibition of the SDF-1α-CXCR4 axis by the CXCR4 antagonist AMD3100 suppresses the migration of cultured cells from ATL patients and murine lymphoblastoid cells from HTLV-I Tax transgenic mice [J]. Blood , 2009 , 114 ( 14 ): 2961 - 2968 .
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