少腹逐瘀汤对子宫内膜异位症大鼠MAPK/ERK信号通路的影响
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摘要
目的:观察少腹逐瘀汤对子宫内膜异位症大鼠丝裂原激活蛋白激酶(MAPK)/细胞外信号调节激酶(ERK)信号通路的影响,探讨少腹逐瘀汤治疗子宫内膜异位症的分子机制。方法:健康雌性SD大鼠随机分为空白对照组、模型组、痛经宝阳性对照组及少腹逐瘀汤低、中、高剂量组。构建大鼠子宫内腹异位症模型,药物处理4周后,HE染色观察异位子宫组织的病理变化; ELISA法检测宫腔组织中肿瘤坏死因子α(TNF-α)、白细胞介素6(IL-6)及IL-8的含量; RT-qPCR检测宫腔组织中ERK、血管内皮生长因子(VEGF)和基质金属蛋白酶9(MMP-9)的mRNA表达量; Western blot检测宫腔组织中核因子-κB(NF-κB)、MAPK和MAPK-ERK激酶(MEK)的蛋白含量。结果:与模型组相比,少腹逐瘀汤中、高剂量组大鼠子宫组织中TNF-α、IL-6及IL-8含量显著降低(P <0. 05),ERK、VEGF和MMP-9的m RNA表达量显著降低(P <0. 05),NF-κB、MEK和MAPK的蛋白表达显著下降(P <0. 05)。结论:少腹逐瘀汤通过调节MAPK/ERK信号通路中关键分子的作用,达到治疗子宫内膜异位症的效果。
AIM: To observe the effects of Shaofu-Zhuyu decoction( SFZY) on mitogen-activated protein kinase( MAPK)/extracellular signal-regulated kinase( ERK) signaling pathway in the rats with endometriosis( EM),and to explore the mechanism of SFZY for treatment of EM. METHODS: Healthy female SD rats were used to establish the EM model. The rats were randomly divided into blank control group,model group,positive control group,and low dose,middle dose and high dose of SFZY groups. The pathological changes of the endometriotic tissue were observed by HE staining.The levels of tumor necrosis factor-α( TNF-α),interleukin-6( IL-6) and IL-8 in the uterine tissue were detected by ELISA. The m RNA expression of ERK,vascular endothelial growh factor( VEGF) and matrix metalloprotein-9( MMP-9)was detected by RT-qPCR. The protein expression of nuclear factor-κB( NF-κB),MAPK and MAPK-ERK kinase( MEK)was determined by Western blot. RESULTS: Compared with model group,the levels of TNF-α,IL-6 and IL-8 in the uterine tissue of the rats in middle dose and high dose of SFZY groups were significantly decreased( P < 0. 05),the m RNA expression of ERK,VEGF and MMP-9 was significantly reduced,and the protein expression of NF-κB,MEK and MAPK was decreased significantly in the rat endometriotic tissues( P < 0. 05). CONCLUSION: SFZY may play a key role in the treatment of EM by regulating MAPK/ERK signaling pathway.
AIM: To observe the effects of Shaofu-Zhuyu decoction( SFZY) on mitogen-activated protein kinase( MAPK)/extracellular signal-regulated kinase( ERK) signaling pathway in the rats with endometriosis( EM),and to explore the mechanism of SFZY for treatment of EM. METHODS: Healthy female SD rats were used to establish the EM model. The rats were randomly divided into blank control group,model group,positive control group,and low dose,middle dose and high dose of SFZY groups. The pathological changes of the endometriotic tissue were observed by HE staining.The levels of tumor necrosis factor-α( TNF-α),interleukin-6( IL-6) and IL-8 in the uterine tissue were detected by ELISA. The m RNA expression of ERK,vascular endothelial growh factor( VEGF) and matrix metalloprotein-9( MMP-9)was detected by RT-qPCR. The protein expression of nuclear factor-κB( NF-κB),MAPK and MAPK-ERK kinase( MEK)was determined by Western blot. RESULTS: Compared with model group,the levels of TNF-α,IL-6 and IL-8 in the uterine tissue of the rats in middle dose and high dose of SFZY groups were significantly decreased( P < 0. 05),the m RNA expression of ERK,VEGF and MMP-9 was significantly reduced,and the protein expression of NF-κB,MEK and MAPK was decreased significantly in the rat endometriotic tissues( P < 0. 05). CONCLUSION: SFZY may play a key role in the treatment of EM by regulating MAPK/ERK signaling pathway.
引文
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[11]谭玉珍,王海杰,张文彩,等.血管内皮生长因子-C促进淋巴管内皮细胞的增殖、迁移和F-肌动蛋白重组[J].解剖学报,2003,34(6):633-637.
[12]Schoppmann SF,Birner P,St`ckl J,et al.Tumor-associated macrophages express lymphatic endothelial growth factors and are related to peritumoral lymphangiogenesis[J].Am J Pathol,2002,161(3):947-956.
[13]LI XF,Charnock-Jones DS,Zhang E,et al.Angiogenic growth factor messenger ribonucleic acids in uterine natural killer cells[J].J Clin Endocrinol Metab,2001,86(4):1823-1834.
[14]董倩,邱晓红.血管生成及抗血管生成与子宫内膜异位症关系研究进展[J].中国实用妇科与产科杂志,2012,28(3):235-237.
[15]夏玉芳.雌、孕激素对子宫内膜异位症裸鼠模型中MMP-2、TIMP-2及VEGF蛋白表达的影响[D].青岛:青岛大学,2007.
[16]Tan XJ,Lang JH,Liu DY,et al.Expression of vascular endothelial growth factor and thrombospondin-1 m RNA in patients with endometriosis[J].Fertil Steril,2002,78(1):148-153.
[17]Collette T,Maheux R,Mailloux J,et al.Increased expression of matrix metalloproteinase-9 in the eutopic endometrial tissue of women with endometriosis[J].Hum Reprod,2006,21(12):3059-3067.
[18]Park JS,Lee JH,Kim M,et al.Endometrium from women with endometriosis shows increased proliferation activity[J].Fertil Steril,2009,92(4):1246-1249.
[19]Yotova IY,Quan P,Leditznig N,et al.Abnormal activation of Ras/Raf/MAPK and Rho A/ROCKII signalling pathways in eutopic endometrial stromal cells of patients with endometriosis[J].Hum Reprod,2011,26(4):885-897.
[20]Ng8 C,Nicco C,Leconte M,et al.Protein kinase inhibitors can control the progression of endometriosis in vitro and in vivo[J].J Pathol,2010,222(2):148-157.
[21]Rosengart MR,Arbabi S,Garcia I,et al.Interactions of calcium/calmodulin-dependent protein kinases(Ca MK)and extracellular-regulated kinase(ERK)in monocyte adherence and TNFαproduction[J].Shock,2000,13(3):183-189.
[22]杨海杰,王勉,王磊,等.IL-6通过MAPK/ERK信号通路调节BMSCs的软骨定向分化[J].中国生物化学与分子生物学报,2015,31(2):153-160.
[23]李建民,陈长香,赵雅宁,等.大鼠睡眠剥夺与p38MAPK信号通路的相关性研究[J].现代预防医学,2011,38(16):3270-3272.
[24]Malik S,Day K,Perrault I,et al.Menstrual effluent in endometriosis shows no difference in volume,VEGF-A,MMP2 and MMP9 or s FLT[J].Reprod Biomed Online,2006,12(2):174-181.
[25]朱虹.MMP-9、VEGF在子宫内膜异位症中的表达及意义[D].南昌:南昌大学,2007.
[2]梁炎春,王伟,黄佳明,等.Sema 3A及其受体在子宫内膜异位症中的表达及意义[J].中国实用妇科与产科杂志,2015,31(5):467-472.
[3]王静,罗小琼,陈升才.子宫内膜异位症疼痛治疗的研究进展[J].现代医药卫生,2017,33(1):83-85.
[4]Huntington A,Gilmour JA.A life shaped by pain:women and endometriosis[J].J Clin Nurs,2010,14(9):1124-1132.
[5]Nanney A,Muro K,Levy RM.Implanted drug delivery systems for the control of chronic pain[M]∥Benzon HT,Raja SN,Liu SS,et al.Essentials of pain medicine.3rd ed.Philadelphia:Saunders,2011:451-461.
[6]王绪颖,陈彦,张振海,等.化学与药效学指标相结合改进痛经宝颗粒中挥发油提取工艺[J].中国实验方剂学杂志,2011,17(9):15-21.
[7]陈景伟,仝瑞晓,范丽洁,等.BALB/c小鼠子宫内膜异位症痛经模型的建立[J].中国实验动物学报,2014,22(1):83-86.
[8]谷青青.少腹逐瘀汤加减治疗子宫内膜异位症40例临床体会[J].河南医学研究,2017,26(5):837-838.
[9]洪佩佩,张玲璐.少腹逐瘀汤加减辅治子宫内膜异位症痛经的疗效观察[J].浙江中医杂志,2014,49(7):488-489.
[10]朱兰,林宝杏,林笑治.良方温经汤治疗子宫内膜异位症临床研究[J].中国中西医结合杂志,2000,20(9):696-697.
[11]谭玉珍,王海杰,张文彩,等.血管内皮生长因子-C促进淋巴管内皮细胞的增殖、迁移和F-肌动蛋白重组[J].解剖学报,2003,34(6):633-637.
[12]Schoppmann SF,Birner P,St`ckl J,et al.Tumor-associated macrophages express lymphatic endothelial growth factors and are related to peritumoral lymphangiogenesis[J].Am J Pathol,2002,161(3):947-956.
[13]LI XF,Charnock-Jones DS,Zhang E,et al.Angiogenic growth factor messenger ribonucleic acids in uterine natural killer cells[J].J Clin Endocrinol Metab,2001,86(4):1823-1834.
[14]董倩,邱晓红.血管生成及抗血管生成与子宫内膜异位症关系研究进展[J].中国实用妇科与产科杂志,2012,28(3):235-237.
[15]夏玉芳.雌、孕激素对子宫内膜异位症裸鼠模型中MMP-2、TIMP-2及VEGF蛋白表达的影响[D].青岛:青岛大学,2007.
[16]Tan XJ,Lang JH,Liu DY,et al.Expression of vascular endothelial growth factor and thrombospondin-1 m RNA in patients with endometriosis[J].Fertil Steril,2002,78(1):148-153.
[17]Collette T,Maheux R,Mailloux J,et al.Increased expression of matrix metalloproteinase-9 in the eutopic endometrial tissue of women with endometriosis[J].Hum Reprod,2006,21(12):3059-3067.
[18]Park JS,Lee JH,Kim M,et al.Endometrium from women with endometriosis shows increased proliferation activity[J].Fertil Steril,2009,92(4):1246-1249.
[19]Yotova IY,Quan P,Leditznig N,et al.Abnormal activation of Ras/Raf/MAPK and Rho A/ROCKII signalling pathways in eutopic endometrial stromal cells of patients with endometriosis[J].Hum Reprod,2011,26(4):885-897.
[20]Ng8 C,Nicco C,Leconte M,et al.Protein kinase inhibitors can control the progression of endometriosis in vitro and in vivo[J].J Pathol,2010,222(2):148-157.
[21]Rosengart MR,Arbabi S,Garcia I,et al.Interactions of calcium/calmodulin-dependent protein kinases(Ca MK)and extracellular-regulated kinase(ERK)in monocyte adherence and TNFαproduction[J].Shock,2000,13(3):183-189.
[22]杨海杰,王勉,王磊,等.IL-6通过MAPK/ERK信号通路调节BMSCs的软骨定向分化[J].中国生物化学与分子生物学报,2015,31(2):153-160.
[23]李建民,陈长香,赵雅宁,等.大鼠睡眠剥夺与p38MAPK信号通路的相关性研究[J].现代预防医学,2011,38(16):3270-3272.
[24]Malik S,Day K,Perrault I,et al.Menstrual effluent in endometriosis shows no difference in volume,VEGF-A,MMP2 and MMP9 or s FLT[J].Reprod Biomed Online,2006,12(2):174-181.
[25]朱虹.MMP-9、VEGF在子宫内膜异位症中的表达及意义[D].南昌:南昌大学,2007.
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