MenSCs移植对大鼠多囊卵巢综合征的影响及其机制
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摘要
目的探讨经血来源干细胞(MenSCs)移植对多囊卵巢综合征(PCOS)大鼠模型的治疗效果。方法成年雌性SD大鼠45只均分为3组(n=15):对照组不作处理;PCOS组肌内注射脱氢表雄酮(DHEA,60 mg/kg)诱导PCOS;移植组肌内注射DHEA,饲养3周后后经尾静脉注射MenSCs悬液。各组饲养5周后处死,取静脉血,酶联免疫吸附试验(ELISA)检测促卵泡激素(FSH)、黄体生成素(LH)、雌二醇(E2)、孕激素(P)、睾酮(T)、泌乳素(PRL)水平。取卵巢组织进行HE染色观察卵巢形态。实时荧光定量检测卵巢组织miRNA-135表达。Western blot检测卵巢组织Bax和Bcl-2蛋白表达。结果相对于对照组,PCOS组的FSH降低,LH和T均升高(P<0.05);相对于PCOS组,移植组的FSH升高,LH和T均降低(P<0.05)。对照组卵泡形态正常,PCOS组卵巢可见较多囊状扩张的卵泡,移植组多囊状扩张的卵泡较PCOS组减少。与对照组比较,PCOS组miRNA-135和Bcl-2蛋白相对表达水平降低,而Bax蛋白相对表达水平升高(P<0.05);与PCOS组比较,移植组miRNA-135和Bcl-2蛋白相对表达水平升高,Bax蛋白相对表达水平降低(P<0.05)。结论经血来源干细胞移植对大鼠PCOS有明显的改善作用,其机制可能通过上调miRNA-135的表达,进而调控Bax/Bcl-2途径来实现。
Objective To investigate the therapeutic effect of transplantation of menstrual stem cells(MenSCs) on rat model of polycystic ovary syndrome(PCOS).MethodsA total of 45 female SD rats were divided into 3 groups. The control group was not treated. The PCOS group was intramuscularly injected with dehydroepiandrosterone(DHEA,60 mg/kg) to induce PCOS. The transplant group was intramuscularly injected with DHEA. After 3 weeks,the MenSCs suspension was injected through the tail vein. After 5 weeks of feeding,venous blood was taken. ELISA kit was used to detect folliclestimulating hormone(FSH),luteinising hormone(LH),estradiol(E2),progesterone(P),testosterone(T) and prolactinemia(PRL) levels. Ovarian tissues were taken for HE staining to observe ovarian morphology. qPCR was used to detect miRNA-135 expression in ovarian tissue. Western blot assay was used to detect the expression of Bax and Bcl-2 proteins in ovarian tissue.ResultsThe FSH level was significantly decreased,and LH and T levels were significantly increased,in the PCOS group than those in the control group(P<0.05). The FSH level was significantly increased,and LH and T levels were significantly decreased in the transplantation group than those in the PCOS group(P<0.05). The follicle morphology was normal in the control group. Follicles with more cystic dilatation in ovary were observed in the PCOS group. The follicles with polycystic expansion were less in the transplantation group than those in the PCOS group. The expressions of miRNA-135 and Bcl-2 protein were significantly decreased in the PCOS group than those in the control group,and the expression of Bcl-2 protein was significantly higher in the PCOS group than that in the control group(P<0.05). The expressions of miRNA-135 and Bcl-2 protein were significantly higher,the expression of Bas was significantly decreased,in the transplantation group than those of PCOS group(P<0.05).ConclusionMenSCs has a significant improvement effect on rat PCOS,which may be achieved by up-regulating the expression of miRNA-135 and then regulating the Bax/Bcl-2 pathway.
Objective To investigate the therapeutic effect of transplantation of menstrual stem cells(MenSCs) on rat model of polycystic ovary syndrome(PCOS).MethodsA total of 45 female SD rats were divided into 3 groups. The control group was not treated. The PCOS group was intramuscularly injected with dehydroepiandrosterone(DHEA,60 mg/kg) to induce PCOS. The transplant group was intramuscularly injected with DHEA. After 3 weeks,the MenSCs suspension was injected through the tail vein. After 5 weeks of feeding,venous blood was taken. ELISA kit was used to detect folliclestimulating hormone(FSH),luteinising hormone(LH),estradiol(E2),progesterone(P),testosterone(T) and prolactinemia(PRL) levels. Ovarian tissues were taken for HE staining to observe ovarian morphology. qPCR was used to detect miRNA-135 expression in ovarian tissue. Western blot assay was used to detect the expression of Bax and Bcl-2 proteins in ovarian tissue.ResultsThe FSH level was significantly decreased,and LH and T levels were significantly increased,in the PCOS group than those in the control group(P<0.05). The FSH level was significantly increased,and LH and T levels were significantly decreased in the transplantation group than those in the PCOS group(P<0.05). The follicle morphology was normal in the control group. Follicles with more cystic dilatation in ovary were observed in the PCOS group. The follicles with polycystic expansion were less in the transplantation group than those in the PCOS group. The expressions of miRNA-135 and Bcl-2 protein were significantly decreased in the PCOS group than those in the control group,and the expression of Bcl-2 protein was significantly higher in the PCOS group than that in the control group(P<0.05). The expressions of miRNA-135 and Bcl-2 protein were significantly higher,the expression of Bas was significantly decreased,in the transplantation group than those of PCOS group(P<0.05).ConclusionMenSCs has a significant improvement effect on rat PCOS,which may be achieved by up-regulating the expression of miRNA-135 and then regulating the Bax/Bcl-2 pathway.
引文
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[8]曾辉锋,王玉霞,杨莹.脂肪干细胞经两种方法移植后在多囊卵巢综合征大鼠体内分布的比较[J].暨南大学学报(自然科学与医学版),2017,38(5):427-432.Zeng HF,Wang YX,Yang Y.Comparison of the distribution of adipose derived stem cells in rats with polycystic ovary syndrome after transplanting by two ways[J].Journal of Jinan University(Natural Science&Medicine Edition),2017,38(5):427-432.doi:10.11778/j.jdxb.2017.05.010.
[9]崔武庚,宋改环.多囊卵巢综合症动物模型研究[J].宁波大学学报(理工版),2012,25(2):126-128.Cui WG,Song GH.Model of animal polycystic ovary syndrome[J].Journal of Ningbo University(Natural Science and Engineering Edition),2012,25(2):126-128.doi:10.3969/j.issn.1001-5132.2012.02.027.
[10]王臻,黄康榕,王月玲,等.经血来源干细胞移植在小鼠卵巢早衰模型中的定位分布[J].西安交通大学学报(医学版),2017,38(6):803-808.Wang Z,Huang KR,Wang YL,et al.Localization of transplanted menstrual-derived stem cells in premature ovarian failure model of mice[J].Journal of Xi’an Jiaotong University(Medical Sciences),2017,38(6):803-808.doi:10.7652/jdyxb201706005.
[11]Bednarska S,Siejka A.The pathogenesis and treatment of polycystic ovary syndrome:What’s new?[J].Adv Clin Exp Med,2017,26(2):359-367.doi:10.17219/acem/59380.
[12]Mujoo K,Pandita RK,Tiwari A,et al.Differentiation of human induced pluripotent or embryonic stem cells decreases the dna damage repair by homologous recombination[J].Stem Cell Reports,2017,9(5):1660-1674.doi:10.1016/j.stemcr.2017.10.002.
[13]Yin Y,Li X,He XT,et al.Leveraging stem cell homing for therapeutic regeneration[J].J Dent Res,2017,96(6):601-609.doi:10.1177/0022034517706070.
[14]Yagi H,Soto-Gutierrez A,Parekkadan B,et al.Mesenchymal stem cells:mechanisms of immunomodulation and homing[J].Cell Transplant,2010,19(6):667-679.doi:10.3727/096368910X508762.
[15]Xie Q,Wang Z,Zhou H,et al.The role of miR-135-modified adipose-derived mesenchymal stem cells in bone regeneration[J].Biomaterials,2016,75:279-294.doi:10.1016/j.biomaterials.2015.10.042.
[16]Chi XX,Zhang T,Chu XL,et al.The regulatory effect of Genistein on granulosa cell in ovary of rat with PCOS through Bcl-2 and BAXsignaling pathways[J].J Vet Med Sci,2018,80(8):1348-1355.doi:10.1292/jvms.17-0001.
[17]Sun Y,Lin Y,Li H,et al.2,5-Hexanedione induces human ovarian granulosa cell apoptosis through BCL-2,BAX,and CASPASE-3 signaling pathways[J].Arch Toxicol,2012,86(2):205-215.doi:10.1007/s00204-011-0745-7.2019-01-152019-03-20
[2]Yau TT,Ng NY,Cheung LP,et al.Polycystic ovary syndrome:a common reproductive syndrome with long-term metabolic consequences[J].Hong Kong Med J,2017,23(6):622-634.doi:10.12809/hkmj176308.
[3]Ring M.Women’s Health:Polycystic ovarian syndrome,menopause,and osteoporosis[J].Prim Care,2017,44(2):377-398.doi:10.1016/j.pop.2017.02.012.
[4]Patel AN,Park E,Kuzman M,et al.Multipotent menstrual blood stromal stem cells:isolation,characterization,and differentiation[J].Cell Transplant,2008,17(3):303-311.
[5]Wang Z,Wang L,Su X,et al.Rational transplant timing and dose of mesenchymal stromal cells in patients with acute myocardial infarction:A meta-analysis of randomized controlled trials[J].Stem Cell Res Ther,2017,8(1):21.doi:10.1186/s13287-016-0450-9.
[6]Yao Y,Huang C,Gu P,et al.Combined MSC-secreted factors and neural stem cell transplantation promote functional recovery of PDrats[J].Cell Transplant,2016,25(6):1101-1113.doi:10.3727/096368915X689938.
[7]Wang JW,Qiu YR,Fu Y,et al.Transplantation with hypoxiapreconditioned mesenchymal stem cells suppresses brain injurycaused by cardiac arrest-induced global cerebral ischemia in rats[J].J Neurosci Res,2017,95(10):2059-2070.doi:10.1002/jnr.24025.
[8]曾辉锋,王玉霞,杨莹.脂肪干细胞经两种方法移植后在多囊卵巢综合征大鼠体内分布的比较[J].暨南大学学报(自然科学与医学版),2017,38(5):427-432.Zeng HF,Wang YX,Yang Y.Comparison of the distribution of adipose derived stem cells in rats with polycystic ovary syndrome after transplanting by two ways[J].Journal of Jinan University(Natural Science&Medicine Edition),2017,38(5):427-432.doi:10.11778/j.jdxb.2017.05.010.
[9]崔武庚,宋改环.多囊卵巢综合症动物模型研究[J].宁波大学学报(理工版),2012,25(2):126-128.Cui WG,Song GH.Model of animal polycystic ovary syndrome[J].Journal of Ningbo University(Natural Science and Engineering Edition),2012,25(2):126-128.doi:10.3969/j.issn.1001-5132.2012.02.027.
[10]王臻,黄康榕,王月玲,等.经血来源干细胞移植在小鼠卵巢早衰模型中的定位分布[J].西安交通大学学报(医学版),2017,38(6):803-808.Wang Z,Huang KR,Wang YL,et al.Localization of transplanted menstrual-derived stem cells in premature ovarian failure model of mice[J].Journal of Xi’an Jiaotong University(Medical Sciences),2017,38(6):803-808.doi:10.7652/jdyxb201706005.
[11]Bednarska S,Siejka A.The pathogenesis and treatment of polycystic ovary syndrome:What’s new?[J].Adv Clin Exp Med,2017,26(2):359-367.doi:10.17219/acem/59380.
[12]Mujoo K,Pandita RK,Tiwari A,et al.Differentiation of human induced pluripotent or embryonic stem cells decreases the dna damage repair by homologous recombination[J].Stem Cell Reports,2017,9(5):1660-1674.doi:10.1016/j.stemcr.2017.10.002.
[13]Yin Y,Li X,He XT,et al.Leveraging stem cell homing for therapeutic regeneration[J].J Dent Res,2017,96(6):601-609.doi:10.1177/0022034517706070.
[14]Yagi H,Soto-Gutierrez A,Parekkadan B,et al.Mesenchymal stem cells:mechanisms of immunomodulation and homing[J].Cell Transplant,2010,19(6):667-679.doi:10.3727/096368910X508762.
[15]Xie Q,Wang Z,Zhou H,et al.The role of miR-135-modified adipose-derived mesenchymal stem cells in bone regeneration[J].Biomaterials,2016,75:279-294.doi:10.1016/j.biomaterials.2015.10.042.
[16]Chi XX,Zhang T,Chu XL,et al.The regulatory effect of Genistein on granulosa cell in ovary of rat with PCOS through Bcl-2 and BAXsignaling pathways[J].J Vet Med Sci,2018,80(8):1348-1355.doi:10.1292/jvms.17-0001.
[17]Sun Y,Lin Y,Li H,et al.2,5-Hexanedione induces human ovarian granulosa cell apoptosis through BCL-2,BAX,and CASPASE-3 signaling pathways[J].Arch Toxicol,2012,86(2):205-215.doi:10.1007/s00204-011-0745-7.2019-01-152019-03-20