生长激素和全反式维甲酸拮抗子宫内膜纤维化的机制
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摘要
目的探讨生长激素(GH)和全反式维甲酸(ATRA)拮抗子宫内膜纤维化的机制。方法选取新鲜子宫内膜组织进行原代人子宫内膜基质细胞(ESCs)培养。实验细胞分别设立正常组、模型组、GH实验组、ATRA实验组。用逆转录聚合酶链反应(RT-PCR)测定细胞骨桥蛋白(OPN)、整合素αγβ3(Itgαγβ3)、Ⅰ型胶原α1(COL1A1)、α-平滑肌肌动蛋白(α-SMA)mRNA的表达水平,用免疫蛋白印迹试验(Western blot)测定细胞中OPN、Itgαγβ3、COL1A1、α-SMA蛋白表达水平。结果与正常组比较,模型组中细胞的Itgαγβ3和OPN mRNA水平降低(P<0.05);与模型组比较,GH实验组细胞的OPN、ItgαγβmRNA表达水平增加(P<0.05),且与正常组比较,差异无统计学意义(P>0.05)。与正常组比较,模型组COL1A1、α-SMA mRNA的表达量显著增加(P<0.05);与模型组比较,ATRA实验组中细胞COL1A1、α-SMA mRNA的表达量减少(P<0.05),且与正常组比较,差异无统计学意义(P>0.05)。与正常组比较,模型组中细胞OPN、Itgαγβ3蛋白水平降低(P<0.05);与模型组比较,GH实验组细胞中OPN、Itgαγβ蛋白表达水平增加(P<0.05),且与正常组比较,差异无统计学意义(P>0.05)。与正常组比较,模型组中COL1A1、α-SMA蛋白的表达水平增加,与模型组比较,ATRA实验组中细胞COL1A1、α-SMA蛋白的表达水平下调(P<0.05),且与正常组比较,差异无统计学意义(P>0.05)。结论 GH和ATRA可分别通过促进OPN、Itgαγβ3mRNA的表达及抑制COL1A1、α-SMA mRNA的表达来拮抗子宫内膜纤维化。
Objective To study the mechanism of growth hormone(GH)and all-trans retinoic acid in antagonizing endometrial fibrosis.Methods Fresh endometrial tissue was selected to culture primary human endometrial stromal cells(ESCs).The experimental cells were divided into normal group,model control group,GH test group and ATRA test group.The normal group did not use drug intervention,the model group intervened 48 hwith 10 ng/mL TGF-beta 1,and the GH experimental group was treated with 10 ng/mL TGF-beta1 after 48 hto add 48 hto 0.1 mol/L GH,and the ATRA experimental group was treated with 10 ng/mL.The levels of OPN,Itgαγβ3,COL1 A1,andα-SMA mRNA were measured by RT-PCR.The Western blot method was used to determine the expression of OPN,Itgαγβ3,COL1 A1 and α-SMA protein in the cells.Results Compared with the normal control group,the OPN and Itgαγβ3 mRNA levels in the model group were significantly decreased(P<0.05).The expression levels of OPN and Itgαγβin the GH group were higher than those in the model group(P<0.05),and no significant difference was found when compared with the normal group(P>0.05).Compared with the normal group,the expression of COL1 A1 andα-SMA mRNA in the model group was significantly increased(P<0.05).Compared with the model group,the expression of COL1 A1 andα-SMA mRNA in the experimental group of all-trans retinoic acid was significantly reduced(P<0.05),and no significant difference was found when compared with the normal group(P>0.05).Compared with the normal control group,the OPN and Itgαγβ3 of the cells in the model group were significantly decreased(P<0.05).Compared with the model group,the expression levels of OPN and Itgαγβin the GH group were significantly increased(P<0.05),and no significant difference was found when compared with the normal group(P>0.05).Compared with the normal group,the COL1 A1 andα-SMA proteins in the model group was significantly down-regulated in the all-trans retinoic acid group(P<0.05),and there was no significant difference with the normal group(P>0.05).Conclusion GH and all-trans retinoic acid can antagonize endometrial fibrosis by promoting the expression of OPN and Itgαγβ3 mRNA and inhibiting the expression of COL1 A1 andα-SMA mRNA.
Objective To study the mechanism of growth hormone(GH)and all-trans retinoic acid in antagonizing endometrial fibrosis.Methods Fresh endometrial tissue was selected to culture primary human endometrial stromal cells(ESCs).The experimental cells were divided into normal group,model control group,GH test group and ATRA test group.The normal group did not use drug intervention,the model group intervened 48 hwith 10 ng/mL TGF-beta 1,and the GH experimental group was treated with 10 ng/mL TGF-beta1 after 48 hto add 48 hto 0.1 mol/L GH,and the ATRA experimental group was treated with 10 ng/mL.The levels of OPN,Itgαγβ3,COL1 A1,andα-SMA mRNA were measured by RT-PCR.The Western blot method was used to determine the expression of OPN,Itgαγβ3,COL1 A1 and α-SMA protein in the cells.Results Compared with the normal control group,the OPN and Itgαγβ3 mRNA levels in the model group were significantly decreased(P<0.05).The expression levels of OPN and Itgαγβin the GH group were higher than those in the model group(P<0.05),and no significant difference was found when compared with the normal group(P>0.05).Compared with the normal group,the expression of COL1 A1 andα-SMA mRNA in the model group was significantly increased(P<0.05).Compared with the model group,the expression of COL1 A1 andα-SMA mRNA in the experimental group of all-trans retinoic acid was significantly reduced(P<0.05),and no significant difference was found when compared with the normal group(P>0.05).Compared with the normal control group,the OPN and Itgαγβ3 of the cells in the model group were significantly decreased(P<0.05).Compared with the model group,the expression levels of OPN and Itgαγβin the GH group were significantly increased(P<0.05),and no significant difference was found when compared with the normal group(P>0.05).Compared with the normal group,the COL1 A1 andα-SMA proteins in the model group was significantly down-regulated in the all-trans retinoic acid group(P<0.05),and there was no significant difference with the normal group(P>0.05).Conclusion GH and all-trans retinoic acid can antagonize endometrial fibrosis by promoting the expression of OPN and Itgαγβ3 mRNA and inhibiting the expression of COL1 A1 andα-SMA mRNA.
引文
[1]李小玉.生长激素对子宫内膜异位症患者卵巢颗粒细胞凋亡的影响[D].石家庄:河北医科大学,2016.
[2]CHRISTIANSEN J S,BACKELJAUW P F,BIDLING-MAIER M,et al.Growth hormone research society perspective on the development of long-acting growth hormone preparations[J].Eur J Endocrinol,2016,174(6):C1-C8.
[3]何鹏,周青山.重组人生长激素对老年胸外伤患者蛋白质代谢及免疫功能的影响[J].贵阳医学院学报,2015,40(3):307-309.
[4]LI X L,WANG L,LV F,et al.The influence of different growth hormone addition protocols to poor ovarian responders on clinical outcomes in controlled ovary stimulation cycles:a systematic review and meta-analysis[J].Medicine(Baltimore),2017,96(12):e6443.
[5]张磊,陈秀萍,蒋玲,等.全反式维甲酸对肾小球硬化大鼠肾小球TRPC6表达的影响[J].实用医学杂志,2016,32(23):3827-3831.
[6]李杏婵,梁翠霞,李晶,等.宫腔黏连分离术联合生长激素对子宫内膜血流、容积及Smad2/3表达的影响[J].海南医学院学报,2016,22(19):2309-2311.
[7]汤伟钳,胡晓.腭突间充质细胞中TGFβ1信号通路介导维甲酸致腭裂的实验研究[J].中国实用医药,2016,11(32):191-193.
[8]董千靖,段华,汪沙.肌成纤维细胞在子宫内膜异位症纤维化形成中的作用[J].中华妇产科杂志,2017,52(10):714-716.
[9]TAGLIAFERRI D,DE ANGELIS M T,RUSSO N A,et al.Retinoic Acid Specifically Enhances Embryonic Stem Cell Metastate Marked by Zscan4[J].PLoS One,2016,11(2):e0147683.
[10]朱洁茹,欧建平.生长激素对卵巢功能影响的研究进展[J].生殖与避孕,2016,36(6):489-494.
[11]刘志华,王圆.重组人生长激素超促排卵对卵巢反应不良患者的影响分析[J].中国实用医药,2017,12(31):95-97.
[12]代聪伟.IGF-1和IGF-2及其受体对子宫内膜癌细胞生长的影响及机制研究[D].石家庄:河北医科大学,2017.
[13]马新想,孙莹璞,苏迎春,等.生长激素在子宫内膜发育不良中的作用[J].现代妇产科进展,2009,18(5):330-332.
[14]邵晓平,彭程飞,田孝祥,等.全反式维甲酸联合Ⅳ型胶原促进胚胎干细胞向平滑肌细胞分化[J].中国动脉硬化杂志,2016,24(2):119-123.
[15]SHAROVA L V,SHAROV A A,PIAO Y,et al.Emergence of undifferentiated colonies from mouse embryonic stem cells undergoing differentiation by retinoic acid treatment[J].In Vitro Cell Dev Biol Anim,2016,52(5):616-624.
[16]胡海燕.全反式维甲酸在宫腔粘连模型中抗子宫内膜纤维化作用及机制研究[D].广州:南方医学大学,2017.
[17]鄢力,苟鹏,唐芳,等.重组人生长激素对生长激素缺乏症患儿部分相关激素水平的影响[J].贵州医科大学学报,2017,42(5):557-560.
[18]SUBRAMANIAN U,NAGARAJAN D.All-Trans retinoic acid supplementation prevents cardiac fibrosis and cytokines induced by methylglyoxal[J].Glycoconj J,2017,34(2):255-265.
[19]LEEM A Y,SHIN M H,DOUGLAS I S,et al.All-trans retinoic acid attenuates bleomycin-induced pulmonary fibrosis via downregulating EphA2-EphrinAl signaling[J].Biochem Biophys Res Commun,2017,491(3):721-726.
[20]PRIYANKA S H,SYAM S D,NAIR S S,et al.All-trans retinoic acid modulates TNF-αand CYP2E1 pathways and enhances regression of ethanol-induced fibrosis markers in hepatocytes and HSCs in abstaining rodent model[J].Arch Physiol Biochem,2018:1-9.
[21]刘霞,马红萍.全反式维甲酸对5/6肾大部切除大鼠肾间质纤维化的影响及其作用机制[J].复旦学报(医学版),2016,43(1):30-35.
[22]ZHANG S,SHI R,CHEN S,et al.All-trans retinoic acid inhibits the proliferation of SGC7901cells by regulating caveolin-1 localization via the ERK/MAPK signaling pathway[J].Oncol Lett,2018,15(2):1523-1528.
[23]CHEN X,QIN Y,ZHOU T,et al.The potential role of retinoic acid receptor alpha on glomerulosclerosis in rats and podocytes injury is associated with the induction of MMP2 and MMP9[J].Acta Biochim Biophys Sin(Shanghai),2017,49(8):669-679.
[2]CHRISTIANSEN J S,BACKELJAUW P F,BIDLING-MAIER M,et al.Growth hormone research society perspective on the development of long-acting growth hormone preparations[J].Eur J Endocrinol,2016,174(6):C1-C8.
[3]何鹏,周青山.重组人生长激素对老年胸外伤患者蛋白质代谢及免疫功能的影响[J].贵阳医学院学报,2015,40(3):307-309.
[4]LI X L,WANG L,LV F,et al.The influence of different growth hormone addition protocols to poor ovarian responders on clinical outcomes in controlled ovary stimulation cycles:a systematic review and meta-analysis[J].Medicine(Baltimore),2017,96(12):e6443.
[5]张磊,陈秀萍,蒋玲,等.全反式维甲酸对肾小球硬化大鼠肾小球TRPC6表达的影响[J].实用医学杂志,2016,32(23):3827-3831.
[6]李杏婵,梁翠霞,李晶,等.宫腔黏连分离术联合生长激素对子宫内膜血流、容积及Smad2/3表达的影响[J].海南医学院学报,2016,22(19):2309-2311.
[7]汤伟钳,胡晓.腭突间充质细胞中TGFβ1信号通路介导维甲酸致腭裂的实验研究[J].中国实用医药,2016,11(32):191-193.
[8]董千靖,段华,汪沙.肌成纤维细胞在子宫内膜异位症纤维化形成中的作用[J].中华妇产科杂志,2017,52(10):714-716.
[9]TAGLIAFERRI D,DE ANGELIS M T,RUSSO N A,et al.Retinoic Acid Specifically Enhances Embryonic Stem Cell Metastate Marked by Zscan4[J].PLoS One,2016,11(2):e0147683.
[10]朱洁茹,欧建平.生长激素对卵巢功能影响的研究进展[J].生殖与避孕,2016,36(6):489-494.
[11]刘志华,王圆.重组人生长激素超促排卵对卵巢反应不良患者的影响分析[J].中国实用医药,2017,12(31):95-97.
[12]代聪伟.IGF-1和IGF-2及其受体对子宫内膜癌细胞生长的影响及机制研究[D].石家庄:河北医科大学,2017.
[13]马新想,孙莹璞,苏迎春,等.生长激素在子宫内膜发育不良中的作用[J].现代妇产科进展,2009,18(5):330-332.
[14]邵晓平,彭程飞,田孝祥,等.全反式维甲酸联合Ⅳ型胶原促进胚胎干细胞向平滑肌细胞分化[J].中国动脉硬化杂志,2016,24(2):119-123.
[15]SHAROVA L V,SHAROV A A,PIAO Y,et al.Emergence of undifferentiated colonies from mouse embryonic stem cells undergoing differentiation by retinoic acid treatment[J].In Vitro Cell Dev Biol Anim,2016,52(5):616-624.
[16]胡海燕.全反式维甲酸在宫腔粘连模型中抗子宫内膜纤维化作用及机制研究[D].广州:南方医学大学,2017.
[17]鄢力,苟鹏,唐芳,等.重组人生长激素对生长激素缺乏症患儿部分相关激素水平的影响[J].贵州医科大学学报,2017,42(5):557-560.
[18]SUBRAMANIAN U,NAGARAJAN D.All-Trans retinoic acid supplementation prevents cardiac fibrosis and cytokines induced by methylglyoxal[J].Glycoconj J,2017,34(2):255-265.
[19]LEEM A Y,SHIN M H,DOUGLAS I S,et al.All-trans retinoic acid attenuates bleomycin-induced pulmonary fibrosis via downregulating EphA2-EphrinAl signaling[J].Biochem Biophys Res Commun,2017,491(3):721-726.
[20]PRIYANKA S H,SYAM S D,NAIR S S,et al.All-trans retinoic acid modulates TNF-αand CYP2E1 pathways and enhances regression of ethanol-induced fibrosis markers in hepatocytes and HSCs in abstaining rodent model[J].Arch Physiol Biochem,2018:1-9.
[21]刘霞,马红萍.全反式维甲酸对5/6肾大部切除大鼠肾间质纤维化的影响及其作用机制[J].复旦学报(医学版),2016,43(1):30-35.
[22]ZHANG S,SHI R,CHEN S,et al.All-trans retinoic acid inhibits the proliferation of SGC7901cells by regulating caveolin-1 localization via the ERK/MAPK signaling pathway[J].Oncol Lett,2018,15(2):1523-1528.
[23]CHEN X,QIN Y,ZHOU T,et al.The potential role of retinoic acid receptor alpha on glomerulosclerosis in rats and podocytes injury is associated with the induction of MMP2 and MMP9[J].Acta Biochim Biophys Sin(Shanghai),2017,49(8):669-679.