來氟米特治療子宮內(nèi)膜異位癥大鼠模型的實驗研究
發(fā)布時間：2018-08-10 20:44
【摘要】：第一部分 非動情期自體移植法子宮內(nèi)膜異位癥大鼠模型的建立 目的：用自體子宮內(nèi)膜移植法構(gòu)建子宮內(nèi)膜異位癥大鼠模型,為進一步的實驗奠定基礎(chǔ)。 方法：健康雌性未孕SD大鼠48只,周齡8-10周,體重200-250g。參考Vernon等的方法進行自體子宮內(nèi)膜移植。建模后3周進行第二次剖腹探查,觀察子宮內(nèi)膜種植生長情況和組織形態(tài)學(xué)變化。移植灶體積增大,呈透明結(jié)節(jié)狀或囊狀,內(nèi)有淡黃色透明囊液積聚者視為造模成功。 結(jié)果：共做48只SD大鼠模型,有40例大鼠在移植處可見透明小囊腫,表面有血管形成。SD大鼠的內(nèi)異癥模型成模率為83.33%(40/48),有4例大鼠沒有成模,考慮在取內(nèi)膜片段時內(nèi)膜面積太小或內(nèi)膜與漿肌層分離不夠有關(guān),有2例大鼠死于麻醉意外,還有2例死于腹腔膿腫。其中有10例大鼠異位病灶處腹膜與大網(wǎng)膜粘連,有8例大鼠異位病灶與腸管粘連,余內(nèi)異癥病灶結(jié)構(gòu)清晰,與周圍組織無粘連。 結(jié)論：非動情期自體子宮內(nèi)膜移植法建立子宮內(nèi)膜異位癥大鼠模型是可行的。旨在通過這種建模方法建立理想的子宮內(nèi)膜異位癥動物模型,為臨床治療子宮內(nèi)膜異位癥提供實驗?zāi)Ｐ汀?第二部分 來氟米特治療子宮內(nèi)膜異位癥大鼠模型的實驗研究 目的：初步探討來氟米特對EMT大鼠模型異位灶中MAPK及NF-κB的影響,采用酶聯(lián)免疫法(enzyme-linked immnuno sorbent assay, ELISA)測定大鼠血清中工L-1β的含量,同時觀察其對EMT大鼠異位灶體積的抑制作用及對異位內(nèi)膜病理形態(tài)學(xué)的影響,并對其可能的機制進行探討。進一步證實免疫調(diào)節(jié)劑治療子宮內(nèi)膜異位癥是可行的,為子宮內(nèi)膜異位癥的臨床治療提供新的治療策略和靶點以及實驗依據(jù)。 方法： 1.模型建立3周后,首先抽取實驗動物的尾靜脈血,用ELISA法檢測血清中IL-1β的水平;接著剖腹觀察異位子宮內(nèi)膜的生長情況,測量移植內(nèi)膜病灶的大小,根據(jù)公式體積V=0.52×長×寬×高,計算體積V1,同時把造模成功的40只大鼠隨機分為2組：LEF組(n=20)：給予來氟米特35mg/kg/d灌胃,每天同一時間給藥,連續(xù)用藥3周；模型對照組(n=20)：給予生理鹽水lml/kg/d灌胃,每天同一時間給藥,連續(xù)用藥3周；給藥期間觀察大鼠的活動、飲食、排便、及體重變化等情況。 2.喂藥3周后,再次抽取實驗大鼠的尾靜脈血,并剖腹觀察大鼠移植內(nèi)膜病灶的生長情況并測量異位內(nèi)膜病灶的體積V2,收集移植內(nèi)膜病灶后處死大鼠：(1)V2與V1進行比較；(2)用免疫組化檢測大鼠模型異位灶MAPK及NF-κB的含量；(3)采用ELISA測定大鼠血清中IL-1β的含量；(4)比較各組大鼠治療前后的體重。 結(jié)果： 1.移植模型復(fù)制成功,造模成功率83.33%。移植灶體積增大,呈透明結(jié)節(jié)狀或囊狀,內(nèi)有淡黃色透明囊液積聚。 2.用藥后LEF組移植內(nèi)膜病灶體積(54.18±17.67)小于模型對照組(98.03±9.74),差異有統(tǒng)計學(xué)意義(P0.05)。 3.用藥后LEF組中大鼠移植內(nèi)膜病灶中MAPK和NF-κB免疫組化平均積分光密度值均較模型對照組低,差異有統(tǒng)計學(xué)意義(P0.05)。 4.用藥后LEF組血清IL-1β含量明低于模型對照組,差異有統(tǒng)計學(xué)意義(P0.05);用藥后LEF組血清IL-1β含量明低于用藥前,差異有統(tǒng)計學(xué)意義(P0.05)。 5.用藥后LEF組大鼠行為活躍,毛發(fā)光澤度及飲食方面與模型對照組相比均無明顯改變。且兩組大鼠的體重治療前后相比,差異無統(tǒng)計學(xué)意義(P0.05)。 結(jié)論： 1.LEF可使大鼠移植內(nèi)膜病灶中MAPK和NF-κB的含量降低,使血清IL-1β含量降低。 2.LEF可能通過阻斷EMT大鼠模型的MAPK信號轉(zhuǎn)導(dǎo)通路,調(diào)控NF-κB通路,影響IL-1β的表達和調(diào)控,從而抑制EMT大鼠模型異位子宮內(nèi)膜的發(fā)生及發(fā)展,這可能是其治療EMT的機制之一。
[Abstract]:Part one
Establishment of a rat model of endometriosis induced by non estrous autotransplantation
Objective: To establish a rat model of endometriosis by autologous endometrial transplantation and lay a foundation for further experiment.
Methods: 48 healthy female sterile SD rats, aged 8-10 weeks and weighing 200-250g, underwent autologous endometrial transplantation with reference to Vernon et al. The second exploratory laparotomy was carried out 3 weeks after the establishment of the model. The growth and histomorphological changes of endometrial implantation were observed. Cystic fluid accumulation is regarded as a successful model.
Results: A total of 48 SD rat models were made, 40 of which showed small transparent cysts and angiogenesis on the surface of the graft. The modeling rate of endometriosis in SD rats was 83.33% (40/48), and 4 rats were not. Considering that the intimal area was too small or the intimal membrane was not separated from the serosomal layer, 2 rats died of anesthesia accident. Among them, 10 rats had adhesion between peritoneum and omentum, 8 rats had adhesion between ectopic lesion and intestine, and the remaining endometriosis had clear structure and no adhesion with surrounding tissues.
CONCLUSION: It is feasible to establish a rat model of endometriosis by autologous endometrial transplantation in the non-estrous period.
The second part
Experimental study of leflunomide in the treatment of endometriosis in rats
AIM: To investigate the effects of leflunomide on MAPK and NF-kappa B in ectopic foci of EMT rats, and to determine the content of working L-1 beta in serum by enzyme-linked immnuno sorbent assay (ELISA), and to observe the inhibition of leflunomide on the volume of ectopic foci and the pathomorphological changes of ectopic endometrium in EMT rats. The possible mechanism is discussed. It is further proved that immunomodulators are feasible in the treatment of endometriosis and provide new therapeutic strategies, targets and experimental basis for the clinical treatment of endometriosis.
Method:
1. Three weeks after the establishment of the model, the tail vein blood of the experimental animals was firstly extracted and the level of IL-1 beta in the serum was detected by ELISA. Then the growth of ectopic endometrium was observed by laparotomy and the size of the transplanted endometrial lesion was measured. According to the formula volume V = 0.52 * length * width * height, the volume V1 was calculated. At the same time, 40 successful rats were randomly divided into two groups. LEF group (n=20): Leflunomide 35 mg/kg/d was given intragastric administration at the same time every day for 3 weeks; model control group (n=20): normal saline lml/kg/d was given intragastric administration at the same time every day for 3 weeks; rats'activities, diet, defecation and weight changes were observed during the administration.
2. After 3 weeks, the tail vein blood of experimental rats was extracted again, and the growth of endometrial lesions was observed by laparotomy, and the volume of ectopic endometrial lesions was measured by V2. The rats were sacrificed after endometrial lesions were collected: (1) V2 and V1 were compared; (2) The contents of MAPK and NF-kappa B in the ectopic lesions of the rat model were detected by immunohistochemistry; (3) ELIS was used to detect the volume of ectopic endometrial lesions. A was used to determine the content of IL-1 beta in rat serum; (4) the weight of rats in each group was compared before and after treatment.
Result:
1. The transplantation model was successfully reproduced with a success rate of 83.33%. The volume of the graft was enlarged, with clear nodular or cystic shape and light yellow cystic fluid accumulation.
2. The volume of transplanted endometrial lesions in LEF group was smaller than that in model control group (54.18
3. The mean integral optical density of MAPK and NF-kappa B in LEF group was lower than that in model control group (P 0.05).
4. The level of serum IL-1beta in LEF group was significantly lower than that in model control group (P 0.05), and the level of serum IL-1beta in LEF group was significantly lower than that before treatment (P 0.05).
5. After treatment, the behavior of LEF rats was active, and there were no significant changes in hair gloss and diet compared with the model control group. There was no significant difference in body weight between the two groups before and after treatment (P 0.05).
Conclusion:
1. LEF can decrease the contents of MAPK and NF-kappa B in endometrial lesions of rats, and decrease the content of IL-1 beta in serum.
2. LEF may inhibit the development of ectopic endometrium in EMT rats by blocking MAPK signal transduction pathway, regulating NF-kappa B pathway, affecting the expression and regulation of IL-1 beta, which may be one of the mechanisms of LEF in treating EMT.
【學(xué)位授予單位】：山西醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2013
【分類號】：R-332;R711.71
本文編號：2176152
[Abstract]:Part one
Establishment of a rat model of endometriosis induced by non estrous autotransplantation
Objective: To establish a rat model of endometriosis by autologous endometrial transplantation and lay a foundation for further experiment.
Methods: 48 healthy female sterile SD rats, aged 8-10 weeks and weighing 200-250g, underwent autologous endometrial transplantation with reference to Vernon et al. The second exploratory laparotomy was carried out 3 weeks after the establishment of the model. The growth and histomorphological changes of endometrial implantation were observed. Cystic fluid accumulation is regarded as a successful model.
Results: A total of 48 SD rat models were made, 40 of which showed small transparent cysts and angiogenesis on the surface of the graft. The modeling rate of endometriosis in SD rats was 83.33% (40/48), and 4 rats were not. Considering that the intimal area was too small or the intimal membrane was not separated from the serosomal layer, 2 rats died of anesthesia accident. Among them, 10 rats had adhesion between peritoneum and omentum, 8 rats had adhesion between ectopic lesion and intestine, and the remaining endometriosis had clear structure and no adhesion with surrounding tissues.
CONCLUSION: It is feasible to establish a rat model of endometriosis by autologous endometrial transplantation in the non-estrous period.
The second part
Experimental study of leflunomide in the treatment of endometriosis in rats
AIM: To investigate the effects of leflunomide on MAPK and NF-kappa B in ectopic foci of EMT rats, and to determine the content of working L-1 beta in serum by enzyme-linked immnuno sorbent assay (ELISA), and to observe the inhibition of leflunomide on the volume of ectopic foci and the pathomorphological changes of ectopic endometrium in EMT rats. The possible mechanism is discussed. It is further proved that immunomodulators are feasible in the treatment of endometriosis and provide new therapeutic strategies, targets and experimental basis for the clinical treatment of endometriosis.
Method:
1. Three weeks after the establishment of the model, the tail vein blood of the experimental animals was firstly extracted and the level of IL-1 beta in the serum was detected by ELISA. Then the growth of ectopic endometrium was observed by laparotomy and the size of the transplanted endometrial lesion was measured. According to the formula volume V = 0.52 * length * width * height, the volume V1 was calculated. At the same time, 40 successful rats were randomly divided into two groups. LEF group (n=20): Leflunomide 35 mg/kg/d was given intragastric administration at the same time every day for 3 weeks; model control group (n=20): normal saline lml/kg/d was given intragastric administration at the same time every day for 3 weeks; rats'activities, diet, defecation and weight changes were observed during the administration.
2. After 3 weeks, the tail vein blood of experimental rats was extracted again, and the growth of endometrial lesions was observed by laparotomy, and the volume of ectopic endometrial lesions was measured by V2. The rats were sacrificed after endometrial lesions were collected: (1) V2 and V1 were compared; (2) The contents of MAPK and NF-kappa B in the ectopic lesions of the rat model were detected by immunohistochemistry; (3) ELIS was used to detect the volume of ectopic endometrial lesions. A was used to determine the content of IL-1 beta in rat serum; (4) the weight of rats in each group was compared before and after treatment.
Result:
1. The transplantation model was successfully reproduced with a success rate of 83.33%. The volume of the graft was enlarged, with clear nodular or cystic shape and light yellow cystic fluid accumulation.
2. The volume of transplanted endometrial lesions in LEF group was smaller than that in model control group (54.18
3. The mean integral optical density of MAPK and NF-kappa B in LEF group was lower than that in model control group (P 0.05).
4. The level of serum IL-1beta in LEF group was significantly lower than that in model control group (P 0.05), and the level of serum IL-1beta in LEF group was significantly lower than that before treatment (P 0.05).
5. After treatment, the behavior of LEF rats was active, and there were no significant changes in hair gloss and diet compared with the model control group. There was no significant difference in body weight between the two groups before and after treatment (P 0.05).
Conclusion:
1. LEF can decrease the contents of MAPK and NF-kappa B in endometrial lesions of rats, and decrease the content of IL-1 beta in serum.
2. LEF may inhibit the development of ectopic endometrium in EMT rats by blocking MAPK signal transduction pathway, regulating NF-kappa B pathway, affecting the expression and regulation of IL-1 beta, which may be one of the mechanisms of LEF in treating EMT.
【學(xué)位授予單位】：山西醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2013
【分類號】：R-332;R711.71
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