内异止痛方对子宫内膜异位症大鼠相关mRNAE-cadherin、mRNAβ-catenin的影响
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摘要
[目的]采用逆转录-聚合酶链反应(RT-PCR)技术检测子宫内膜异位症(EMs)模型大鼠异位病灶组织中β-catenin、E-cadherin基因转录水平,探讨内异止痛方对EMs模型大鼠β-catenin、E-cadherin的影响。[方法]取SD大鼠57只,采用自体移植法构建EMs模型,于4周后开腹观察模型构建是否成功,并取模型大鼠5只,利用苏木精-伊红(HE)染色法对其进行组织病理学评价。将40只造模成功的EMs大鼠,随机分为4组,即模型组、中药高、低剂量组、阳性药物组,每组10只,另设假手术组8只(仅进行开腹切除手术,不进行自体移植)。中药高、低剂量治疗组分别灌胃985.3 mg/(kg·d)和245.8 mg/(kg·d)内异止痛方,阳性药物组灌胃7.2 mg/(kg·d)达那唑,模型组及假手术组灌胃2 mL/(kg·d)生理盐水,各组每天均给药1次,连续用药4周;采用RT-PCR技术检测EMs模型大鼠异位病灶组织中β-catenin、E-cadherin转录水平。[结果]与模型组比较,内异止痛方能够显著升高EMs模型大鼠E-cadherinmRNA并降低β-catenin mRNA转录水平,且中药高剂量组各基因转录水平明显优于低剂量组(P<0.01)。[结论]内异止痛方可通过上调E-cadherin或下调β-catenin基因转录来维持E-cad/β-cat复合体结构和功能的稳定性,而达到加强在位内膜细胞间黏附的作用。
[Objective] The expression of β-catenin and E-cadherin gene in ectopic focus tissue of rats with EMs was detected by RT-PCR technique by constructing rat model of endometriosis. To investigate the therapeutic effect of Neiyi Zhitong Prescription on β-catenin and E-cadherin. [Methods] The 57 SD rats were selected and EMs model was constructed by autologous transplantation. The success of the model construction was observed by laparotomy 4 weeks later,and 5 model rats were selected for histopathological evaluation by HE staining. The 40 EMs rats successfully modeled were randomly divided into 4 groups,model group,high-dose and lowdose Chinese medicine treatment group,and positive drug group,with 10 rats in each group and 8 rats in the sham operation group(open resection only,no autologous transplantation). In the high-dose and low-dose groups of traditional Chinese medicine,985.3 mg/(kg·d) and245.8 mg/(kg·d) by Neiyi Zhitong prescription were given. The positive drug group was given 7.2 mg/(kg·d) danazole by gavage,and the model group and the sham operation group were given 2 mL/(kg·d) normal saline by gavage. Each group was given the drug once a day for 4 weeks. Transcription levels of β-catenin and E-cadherin in ectopic lesion tissues of EMs model rats were detected by RT-PCR.[Results] Compared with the model group,Neiyi Zhitong Prescription can significantly increase the m RNA of E-cadherin and reduce the transcription level of β-catenin mRNA in EMs model rats,and the transcription level of each gene in the high-dose group of traditional Chinese medicine is significantly better than that in the low-dose group(P<0.01). [Conclusion] Neiyi Zhitong Prescription can maintain the structural and functional stability of the E-cad/β-cat complex by up-regulating the transcription of E-cadherin or down-regulating the transcription of β-catenin gene,thus enhancing the adhesion between cells in the endometrium.
[Objective] The expression of β-catenin and E-cadherin gene in ectopic focus tissue of rats with EMs was detected by RT-PCR technique by constructing rat model of endometriosis. To investigate the therapeutic effect of Neiyi Zhitong Prescription on β-catenin and E-cadherin. [Methods] The 57 SD rats were selected and EMs model was constructed by autologous transplantation. The success of the model construction was observed by laparotomy 4 weeks later,and 5 model rats were selected for histopathological evaluation by HE staining. The 40 EMs rats successfully modeled were randomly divided into 4 groups,model group,high-dose and lowdose Chinese medicine treatment group,and positive drug group,with 10 rats in each group and 8 rats in the sham operation group(open resection only,no autologous transplantation). In the high-dose and low-dose groups of traditional Chinese medicine,985.3 mg/(kg·d) and245.8 mg/(kg·d) by Neiyi Zhitong prescription were given. The positive drug group was given 7.2 mg/(kg·d) danazole by gavage,and the model group and the sham operation group were given 2 mL/(kg·d) normal saline by gavage. Each group was given the drug once a day for 4 weeks. Transcription levels of β-catenin and E-cadherin in ectopic lesion tissues of EMs model rats were detected by RT-PCR.[Results] Compared with the model group,Neiyi Zhitong Prescription can significantly increase the m RNA of E-cadherin and reduce the transcription level of β-catenin mRNA in EMs model rats,and the transcription level of each gene in the high-dose group of traditional Chinese medicine is significantly better than that in the low-dose group(P<0.01). [Conclusion] Neiyi Zhitong Prescription can maintain the structural and functional stability of the E-cad/β-cat complex by up-regulating the transcription of E-cadherin or down-regulating the transcription of β-catenin gene,thus enhancing the adhesion between cells in the endometrium.
引文
[1]Jacobson TZ,Duffy J,Barlow DH,et al.Laparoscopic surgery for subfertility associated with endometriosis[J].The Cochrane Library,2016,3,4:CD011031.
[2]李艳,闻姬,陈军,等.子宫内膜异位症有关信号通路的研究进展[J].生殖与避孕,2016,36(2):128-134.
[3]Lee WL,Slutsky AS.Sepsis and endothelial permeability[J].N Engl J Med,2013,363(7):689-691.
[4]郭洁,王月平,张崴,等.补肾调冲方对PCOS患病子宫内膜保护作用研究[J].天津中医药,2017,34(6):403-406.
[2]李艳,闻姬,陈军,等.子宫内膜异位症有关信号通路的研究进展[J].生殖与避孕,2016,36(2):128-134.
[3]Lee WL,Slutsky AS.Sepsis and endothelial permeability[J].N Engl J Med,2013,363(7):689-691.
[4]郭洁,王月平,张崴,等.补肾调冲方对PCOS患病子宫内膜保护作用研究[J].天津中医药,2017,34(6):403-406.