不同治疗方法对大鼠子宫内膜异位症的疗效比较

Comparison of different curative methods on endometriosis in rats

目的探讨亮丙瑞林和重组人干扰素-α-2b对Sprague-Dawley(SD)大鼠子宫内膜异位症(EMs)的疗效。方法 2016年8月至11月,选择50只无特殊病原体(SPF)级健康、性成熟的雌性SD大鼠为研究对象。通过自体子宫内膜移植方法建立大鼠腹壁EMs模型,将建模成功的SD大鼠按照随机数字表法,将其分为实验A组(n=15)、实验B组(n=15)和对照组(n=15)。3组大鼠体重、日龄和健康状况等一般资料比较,差异均无统计学意义(P>0.05)。实验A组大鼠皮下注射重组人干扰素-α-2b,剂量为每次10万IU,每隔48h1次,共计3次;实验B组大鼠皮下注射亮丙瑞林20μg/(kg·d)×28d;对照组大鼠皮下注射生理盐水0.1mL/d×28d。治疗4周后,开腹观察各组大鼠子宫内膜异位病灶生长情况,对比分析3组大鼠治疗前及治疗4周后子宫内膜异位病灶体积变化情况,并采用免疫组织化学染色检测异位子宫内膜组织的血管内皮生长因子(VEGF)蛋白和肿瘤坏死因子(TNF)-α蛋白的表达水平。实验过程中对动物的处置符合动物伦理学要求。结果 (1)50只SD大鼠中,EMs建模成功率为90%(45/50)。(2)治疗4周后,3组大鼠子宫内膜异位病灶体积比较,差异有统计学意义(F=139.332,P<0.001),其中,实验A组、实验B组SD大鼠子宫内膜异位病灶体积均较对照组明显减小,差异有统计学意义(LSD-t=13.460、15.280,P<0.001);实验B组大鼠子宫内膜异位病灶体积较实验A组减小,但差异无统计学意义(LSD-t=1.820,P=0.076)。此外,实验A组、实验B组大鼠子宫内膜异位病灶体积均较治疗前减小,并且差异均有统计学意义(t=9.413、14.545,P<0.001)。(3)治疗4周后,3组SD大鼠血清病灶组织中VEGF蛋白、TNF-α蛋白表达水平比较,差异均有统计学意义(F=224.261、225.391,P<0.001)。实验A组、实验B组大鼠VEGF蛋白及TNF-α蛋白表达水平均明显低于对照组,并且差异均有统计学意义(实验A组vs对照组:LSD-t=17.258、19.121,P<0.001;实验B组vs对照组:LSD-t=19.260、17.552,P<0.001)。结论重组人干扰素-α-2b对SD大鼠EMs模型具有明显的治疗作用,并且疗效与亮丙瑞林相当,下调VEGF、TNF-α的表达水平是其可能作用机制之一。但重组人干扰素-α-2b能否代替亮丙瑞林作为新型药物应用于临床,尚待进一步的大样本动物实验及临床循证医学证据以证实。

Objective To discuss therapeutic effects of recombinant human interferon-α-2b and leuprorelin on rat endometriosis （EMs） model .Methods From August to November ,2016 ,a total of 50 healthy specific pathogen fee （SPF） female Sprague-Dawley （SD） rats were chosen as research subjects .The rat model of abdominal wall EMs was established by autologous uterine transplantation .The successfully established rat EMs model were randomly divided into experimental group A （n=15） ,experimental group B （n= 15） and control group （n= 15）according to random number table . There were no significant differences among three groups in the aspects of basic information （ P〉0.05） . The rats in experimental group A received 3 times of 100000 U of recombinant human interferon-α-2b by subcutaneous injection every 48 h .The experimental group B was daily treated with leuprorelin 20 μg/kg by subcutaneous injection ,a total of 28 days .The control group was daily given subcutaneous injection of saline 0 .1 mL ,a total of 28 days .After the end of 4 weeks treatment ,the growth of ectopic lesions in 3 groups were observed ,and the expression of vascular endothelial growth factor （VEGF） protein and tumor necrosis factor （TNF）-α protein in ectopic endometrium tissues were detected by immunohistochemical method . All experimental protocols were approved by the Institutional Animal Care and Use Committee .Results ① Among 50 EMs rats ,the successful rate of EMs rat establishment model was 90% （45/50） .② After 4 weeks treatment ,there was significant difference in ectopic lesions volume among three groups （F=139 .332 ,P〈0 .001） .Among them ,the ectopic lesions volume of rats in experimental group A and experimental group B were both smaller than that of rats in control goup （LSD-t=13 .460 ,15 .280 ;P〈0 .001）;the ectopic lesions volume of rats in experimental group B was smaller than that of rats in experimental group A ,which had no significant （LSD-t=1 .820 ,P=0 .076） .Besides ,there were significant differences in ectopic volume of rats before and after the treatment in experimental group A and B （t=9 .413 ,14 .545 ;P〈0.001） .③ After 4 weeks treatment ,there were no significant in the expression of VEGF protein and TNF-αprotein among 3 groups （F=224 .261 ,225 .391 ;P〈0 .001） .The expression of VEGF protein and TNF-αprotein of experimental group A and B were significant lower than those of control group （experimental group A vs control group ：LSD-t=17 .258 ,19 .121 ;P〈0 .001 ;experimental group B vs control group ：LSD-t=19 .260 ,17 .552 ;P〈0 .001） .Conclusions Recombinant human interferon-α-2b has therapeutic effect on rats with EMs .Downregulating the expression of VEGF and TNF-α is probably one of the mechanisms .Whether recombinant human interferon-α-2b can replace leuprolide as a new drug in clinical application or not ,we still need large-scale animal experiments and evidence-based medicine to confirm .