补肾活血汤对EMS大鼠体内胚胎着床发育的作用及机理初探
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摘要
子宫内膜异位症由于其发病率逐年增加且以疼痛和不孕为主要临床症状而被称为妇科现代病,在不孕患者中约有25%～35%是由于子宫内膜异位症所致,该病严重影响了育龄妇女的工作、学习及家庭生活的和谐,已成为影响生育最主要的妇科疾病之一。本课题拟通过建立自体移植子宫内膜异位症模型大鼠,观察中药补肾活血汤对模型鼠体内卵泡发育数、活胎数的影响,同时检测免疫、生殖内分泌相关指标的变化,探讨补肾活血汤在调节生殖免疫功能方面的作用及机理,寻找中医补肾活血法治疗EMS不孕的科学依据,充分发挥中医药的治疗特色,以造福于不孕症患者。
1.采用自体移植法复制大鼠子宫内膜异位症模型:取雌性未交配性成熟大鼠,术前雌激素诱导,手术开腹取右侧子宫,将自体子宫内膜种植于皮下筋膜层与腹壁肌层之间,常规关腹,清洁级环境饲养4周。
2.取上述子宫内膜异位症模型大鼠,随机分为模型对照组、阳性药(内美通)组及补肾活血汤的高、中、低剂量组,另设伪手术组作为正常对照。给药8周后,放射免疫检测血清TNF-α、E_2、P水平,免疫组化染色观察VEGF、MVD、PR表达;同时观察卵泡发育水平和补肾活血汤对怀孕率及活胎数的影响,探讨其在调节生殖免疫功能方面的作用及其机理。
1.采用自体移植法成功建立大鼠子宫内膜异位症模型,模型大鼠的TNF-α、E_2水平明显升高,且在位内膜中VEGF、MVD的表达也明显增多,与患者的临床病理改变相仿。
2.整体动物实验结果表明:补肾活血汤各剂量组能显著降低模型大鼠血清中TNF-α、E_2水平,且能减少模型大鼠在位内膜中VEGF、MVD的表达,提示其能通过调节VEGF、TNF-α和MVD水平,抑制异位组织的血管形成来抑制异位病灶的生长,以达到抑制异位子宫内膜的生长、发育;并能提过下调E_2水平改善子宫内膜受容性,促进卵泡发育、增加受精卵着床数目,从而达到提高妊娠率,有效调节生殖功能的目的。
补肾活血汤能有效提高大鼠子宫内膜异位症模型的妊娠功能。其作用有以下几个方面:免疫调节作用;改善机体内分泌水平;改善在位内膜环境;促排卵(可能通过改善下丘脑-垂体-卵巢轴,及卵巢局部的免疫、微循环、代谢微环境,从而诱发排卵)等。
With its incidence rate increasing,Infertility and ache as the main clinical symptoms,endometriosis is called modern gynecology disease,in the sterile patient approximately 25%～35%is due to endometriosis.It has seriously affected the child-bearing age's work of woman,the study and the family life harmony,has become the influence to give birth one of most main gynecology department diseases. Through setting up endometriosis model by rat self-endometria transplantation,this subject was to observe effect of graafian follicle and embryo,immunization and reproduction related index were also detected.To investigate the effects and the mechanisms of Bushen-huoxue decoction(BHD) on regulating reproduction and immune function,searched scientific basis of BHD treating the sterile,displays Chinese medicine treatment characteristic and benefits sterility patient.
1.EMS rats models were established by endometrium autotransplantation: Female virgin rats in estrus stage were induced by estrogen injection prior to operation. During the laparotomy,the right uterus was taken and its endometrium was implanted into between superficial fascia and muscular layer.Turned off the abdomen,the clean level environment raised for 4 weeks.
2.The above model rats were randomized divided into 5 groups:model, high-dose,middle-dose,low-dose and positive drug(Gestrinone).Other sham operated rats were as control group.8 weeks after establishing model,examined by Radioimmunodetection of TNF-α、E_2、P in serum of rats and observed by immunohistochemisty of expression of VEGF、MVD、PR and graafian follicle.The changes of pregnancy rate,embryo was also observed.To study the effects and the mechanisms of BHD on adjust reproduction immunologic function in Endometriosis model Rats.
1.Rat endometriosis models are Successful established by autologous transplantation.The TNF-α、E_2 level of model rats were elevated obviously and the expression of VEGF、MVD in the eutopic endometrial were also increase,It's similar to clinical pathological changes in human.
2.The results of in vivo animal experiments showed that each dose group of BHD could decrease TNF-α、E_2 level in blood serum and the expression of VEGF、MVD in the eutopic endometrial.It showed that it can adjust the VEGF,TNF-αand MVD level,suppress ectopic tissue's vascularization to suppress the dystopia infection's growth,accordingly suppresses the growth,development of dystopia endometrium..By improving endometrium,promotioning Graafian follicle growth, thus achieves raises the rate of pregnancy,adjusts the reproduction function effectively.
The pregnancy function of EMS rats can enhance effectively with BHD:Its function has following several aspects:controlling immunity action;improving organism internal secretion level;improving the eutopic tissues' environment;pressing the ovulation(possibly through to improve hypothalamic-pituitarygland-ovary axis, and ovary partial immunity,microcycle,metabolism micro environment,thus induced ovulation) and so on.
1.采用自体移植法复制大鼠子宫内膜异位症模型:取雌性未交配性成熟大鼠,术前雌激素诱导,手术开腹取右侧子宫,将自体子宫内膜种植于皮下筋膜层与腹壁肌层之间,常规关腹,清洁级环境饲养4周。
2.取上述子宫内膜异位症模型大鼠,随机分为模型对照组、阳性药(内美通)组及补肾活血汤的高、中、低剂量组,另设伪手术组作为正常对照。给药8周后,放射免疫检测血清TNF-α、E_2、P水平,免疫组化染色观察VEGF、MVD、PR表达;同时观察卵泡发育水平和补肾活血汤对怀孕率及活胎数的影响,探讨其在调节生殖免疫功能方面的作用及其机理。
1.采用自体移植法成功建立大鼠子宫内膜异位症模型,模型大鼠的TNF-α、E_2水平明显升高,且在位内膜中VEGF、MVD的表达也明显增多,与患者的临床病理改变相仿。
2.整体动物实验结果表明:补肾活血汤各剂量组能显著降低模型大鼠血清中TNF-α、E_2水平,且能减少模型大鼠在位内膜中VEGF、MVD的表达,提示其能通过调节VEGF、TNF-α和MVD水平,抑制异位组织的血管形成来抑制异位病灶的生长,以达到抑制异位子宫内膜的生长、发育;并能提过下调E_2水平改善子宫内膜受容性,促进卵泡发育、增加受精卵着床数目,从而达到提高妊娠率,有效调节生殖功能的目的。
补肾活血汤能有效提高大鼠子宫内膜异位症模型的妊娠功能。其作用有以下几个方面:免疫调节作用;改善机体内分泌水平;改善在位内膜环境;促排卵(可能通过改善下丘脑-垂体-卵巢轴,及卵巢局部的免疫、微循环、代谢微环境,从而诱发排卵)等。
With its incidence rate increasing,Infertility and ache as the main clinical symptoms,endometriosis is called modern gynecology disease,in the sterile patient approximately 25%～35%is due to endometriosis.It has seriously affected the child-bearing age's work of woman,the study and the family life harmony,has become the influence to give birth one of most main gynecology department diseases. Through setting up endometriosis model by rat self-endometria transplantation,this subject was to observe effect of graafian follicle and embryo,immunization and reproduction related index were also detected.To investigate the effects and the mechanisms of Bushen-huoxue decoction(BHD) on regulating reproduction and immune function,searched scientific basis of BHD treating the sterile,displays Chinese medicine treatment characteristic and benefits sterility patient.
1.EMS rats models were established by endometrium autotransplantation: Female virgin rats in estrus stage were induced by estrogen injection prior to operation. During the laparotomy,the right uterus was taken and its endometrium was implanted into between superficial fascia and muscular layer.Turned off the abdomen,the clean level environment raised for 4 weeks.
2.The above model rats were randomized divided into 5 groups:model, high-dose,middle-dose,low-dose and positive drug(Gestrinone).Other sham operated rats were as control group.8 weeks after establishing model,examined by Radioimmunodetection of TNF-α、E_2、P in serum of rats and observed by immunohistochemisty of expression of VEGF、MVD、PR and graafian follicle.The changes of pregnancy rate,embryo was also observed.To study the effects and the mechanisms of BHD on adjust reproduction immunologic function in Endometriosis model Rats.
1.Rat endometriosis models are Successful established by autologous transplantation.The TNF-α、E_2 level of model rats were elevated obviously and the expression of VEGF、MVD in the eutopic endometrial were also increase,It's similar to clinical pathological changes in human.
2.The results of in vivo animal experiments showed that each dose group of BHD could decrease TNF-α、E_2 level in blood serum and the expression of VEGF、MVD in the eutopic endometrial.It showed that it can adjust the VEGF,TNF-αand MVD level,suppress ectopic tissue's vascularization to suppress the dystopia infection's growth,accordingly suppresses the growth,development of dystopia endometrium..By improving endometrium,promotioning Graafian follicle growth, thus achieves raises the rate of pregnancy,adjusts the reproduction function effectively.
The pregnancy function of EMS rats can enhance effectively with BHD:Its function has following several aspects:controlling immunity action;improving organism internal secretion level;improving the eutopic tissues' environment;pressing the ovulation(possibly through to improve hypothalamic-pituitarygland-ovary axis, and ovary partial immunity,microcycle,metabolism micro environment,thus induced ovulation) and so on.
引文
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