基于雌激素温敏水凝胶混液的子宫内膜异位症小鼠模型的构建

Method for developing mouse model of endometrios is based on estrogen-releasing thermo-sensitive hydrogel system

目的探讨并优化基于雌激素温敏水凝胶混液皮下注射的子宫内膜异位症(EM)小鼠模型构建的条件。方法(1)在小鼠皮下分别注射不同浓度梯度的雌激素温敏水凝胶混液,3 d后检测小鼠血雌激素浓度,制作小鼠血雌激素浓度-雌激素温敏水凝胶混液浓度曲线,选择最佳造模雌激素温敏水凝胶混液浓度;(2)在小鼠均皮下注射最佳浓度雌激素温敏水凝胶混液,分别于第1、3、5、7及10天检测小鼠血雌激素浓度,明确雌激素温敏水凝胶混液小鼠皮下注射后的释放规律;(3)将供体小鼠子宫内膜组织移植到9只受体小鼠的腹腔肠系膜上,受体小鼠分成a、b、c组,a组移植后每天腹腔注射PBS,b组移植后每天腹腔注射雌激素溶液,c组移植后每周皮下注射最佳浓度雌激素温敏水凝胶混液。21 d后观察EM小鼠模型的成功率。结果(1)小鼠血雌激素浓度与温敏水凝胶混液中雌激素浓度呈线性关系,0.8 mg/ml为最佳造模雌激素温敏水凝胶混液浓度;(2)每只小鼠按0.8 mg/ml的雌激素温敏水凝胶混液皮下注射0.2 ml后可在小鼠体内释放7 d,稳定维持有效血雌激素浓度;(3)a、b、c组小鼠均存活;b、c组小鼠均见异位囊肿形成,a组小鼠未见明显内异囊肿形成。结论小鼠EM模型的构建需要外源性雌激素的支持。新型缓释剂温敏水凝胶承载雌激素粉末在小鼠皮下注射后,具有持久的缓释效果,雌激素浓度为0.8 mg/ml的温敏水凝胶混液每周皮下注射法支持EM小鼠模型构建的效果明确,值得推广。

Objective This study will explore and optimize the conditions for the construction of the mouse model of endometriosis using the estrogen-releasing thermo-sensitive hydrogel system. Methods (1) The mice were injected subcutaneously with a gradient concentration of estrogen-releasing thermo-sensitive hydrogel systems, and the blood estrogen concentration was measured 3 days later. And then we draw the curve between mouse blood estrogen concentration and thermo-sensitive hydrogel systems concentration, selecting the most appropriate concentration of estrogen-releasing thermo-sensitive hydrogel systems.(2) To explore the release pattern of estrogen-releasing thermo-sensitive hydrogel system, the mice were injected subcutaneously with the optimal concentration of estrogen-releasing thermo-sensitive hydrogel system, and the blood estrogen concentration was measured on the 1st, 3rd, 5th, 7th and 10th day respectively.(3) We transplanted uteruses from donor mice into the abdominal cavities of recipient mice. The recipient mice were divided into three groups named a, b, and c. Group a was intraperitoneally injected with PBS everyday after transplantation, and group b was intraperitoneally injected with estrogen solution everyday after transplantation. And estrogen-releasing thermo-sensitive hydrogel systems mentioned above were injected subcutaneously to group c once a week. Mice were killed 21 days later to observe the success rate of the endometriosis models. Results (1) The concentration of estrogen in mice was linear with the concentration of estrogenin the estrogen-releasing thermo-sensitive hydrogel systems. And 0.8 mg/ml was the most appropriate concentration of the estrogen-releasing thermo-sensitive hydrogel system.(2) The thermo- sensitive hydrogel systems including 0.8 mg/ml estrogen can release estrogen in mice for 7 days and can achieve effective blood estrogen concentration.(3) All mice in group a, b, and c survived. All mice in group b and c showed ectopic cysts, and no cyst was observed in group a. Conclusions The construction of the mouse model of endometriosis requires exogenous estrogen. We have found that the novelestrogen-releasing thermo-sensitive hydrogel systems can slowly release estrogen after subcutaneously injection in mice, and the value of the subcutaneously injection of estrogen-releasing thermo-sensitive hydrogel system (0.8 mg/ml) per week to the construction of the mouse model is certain.The estrogen-releasing thermo-sensitive hydrogel system has high application value and is worthy of promotion.