摘要
目的探讨温肾化瘀法对子宫内膜异位症大鼠妊娠结局的影响及其调控机制,为中药治疗该病提供理论依据。方法将60只雌性大鼠随机分为空白组、模型组、温肾化瘀低剂量组、温肾化瘀高剂量组、西药组,模型组20只,其余组每组10只。除空白组外,其余组大鼠均采用自体子宫内膜组织移植法构建子宫内膜异位症模型。证实造模成功后,温肾化瘀低、高剂量组分别给予1.8 g/kg、5.4 g/kg温肾化瘀方灌胃,西药组给予0.36 mg/kg达那唑灌胃,空白组和模型组均给予2 mL蒸馏水灌胃,均1次/d,连续灌胃4周。灌胃结束后,按照1∶1的比例将雌性大鼠与性成熟的雄性大鼠分笼进行合笼交配,雌鼠出现阴栓时标记为妊娠的第1天。妊娠第4天,各组随机抽取5只大鼠,采用ELISA法检测血清同源框基因A10(HOXA10)和同源框基因A11(HOXA11)、雌激素受体(ER)、孕激素受体(PR)水平,采用Western blot法检测子宫内膜组织中HOXA10、HOXA11、ER、PR蛋白表达情况,采用PCR技术检测子宫内膜组织中HOXA11和白血病抑制因子(LIF)mRNA表达情况;妊娠第20天,统计各组剩余大鼠内异灶面积及大鼠妊娠率、活胎率。结果与空白组比较,模型组血清HOXA10、HOXA11、PR水平和子宫内膜组织中HOXA10、HOXA11蛋白相对表达量及HOXA11 mRNA相对表达量均明显降低(P均<0.05),子宫内膜组织中LIF mRNA相对表达量明显升高(P<0.05)。与模型组比较,温肾化瘀高剂量组血清HOXA10、HOXA11水平和子宫内膜组织中HOXA11、ER、PR蛋白相对表达量及HOXA11 mRNA相对表达量均明显升高(P均<0.05),西药组血清HOXA11水平和子宫内膜组织中HOXA11蛋白及mRNA相对表达量均明显升高(P均<0.05),温肾化瘀低剂量组子宫内膜组织中HOXA11、PR蛋白相对表达量均明显升高(P均<0.05);各药物组子宫内膜组织中LIF mRNA相对表达量均明显降低(P均<0.05)。各药物组大鼠内异灶面积均低于模型组,妊娠率和活胎数量均高于模型组,但差异均无统计学意义(P均>0.05)。结论温肾化瘀法可能通过上调HOXA10、HOXA11、ER、PR表达,下调LIF表达,从而提高子宫内膜容受性,改善妊娠结局。
Objective It is to investigate the effect and its regulatory mechanism of therapy for warming kidney and dissipating blood stasis(WKDBS)on pregnancy outcomes of rats with endometriosis,and to provide some theoretical basis for the treatment of this disease with traditional Chinese medicine.Methods Sixty female rats were randomly divided into blank group,model group,low-dose WKDBS group,high-dose WKDBS group and western medicine group,with 20 rats in the model group and 10 rats in each of the remaining groups.The rats of all groups except for the blank group were used to establish endometriosis models by autologous endometrial tissue transplantation method.After successfully modeling,the low-dose WKDBS group and high-dose WKDBS group were given decoction for WKDBS 1.8 g/kg and 5.4 g/kg by gavage,respectively,and the western medicine group was given danazol 0.36 mg/kg by gavage,while the blank group and the model group were given 2 mL distilled water by gavage,all once daily,continuously treated for 4 weeks.At the end of gavage,the female rats were mated with sexually mature male rats in separate cages in a 1∶1 ratio,and the presence of vaginal plug in females was marked as day 1 of gestation.On the 4th day of pregnancy,5 rats were randomly selected from each group,the serum levels of homeobox A10(HOXA10),homeobox A11(HOXA11),estrogen receptor(ER)and progesterone receptor(PR)were detected by ELISA,and the protein expressions of HOXA10,HOXA11,ER,and PR in endometrial tissues were detected by Western blot,and the mRNA expressions of HOXA11 and leukemia inhibitory factor(LIF)in endometrial tissues were detected by PCR;on the 20th day of pregnancy,the area of endometriosis focus,pregnancy rate and live birth rate of the remaining rats in each group were counted.Results Compared with the blank group,the serum levels of HOXA10,HOXA11,PR and relative expressions of HOXA10,HOXA11 proteins and relative expression of HOXA11 mRNA in endometrial tissues of the model group were significantly decreased(all P<0.05),and relative expression of LIF mRNA in endometrial tissues was significantly increased(P<0.05).Compared with the model group,the serum levels of HOXA10 and HOXA11 and relative expressions of HOXA11,ER and PR proteins as well as relative expression of HOXA11 mRNA in endometrial tissues were significantly increased in the high-dose WKDBS group(all P<0.05),the serum level of HOXA11 and relative expressions of HOXA11 protein and mRNA in endometrial tissues were significantly increased in the western drug group(all P<0.05),and the relative expressions of HOXA11 and PR proteins in endometrial tissue were significantly increased in the low-dose WKDBS group(all P<0.05);the relative expression of LIF mRNA in endometrial tissue were significantly decreased in the treatment groups(all P<0.05).The area of endometriosis focus in rats of each treatment group was lower,while the pregnancy rate and live birth rate were higher than those of the model group,but the differences were not statistically significant(all P>0.05).Conclusion Therapy for warming the kidney and removing blood stasis can increase the endometrial receptivity and improve the pregnancy outcome of rats with endometriosis via up-regulating expressions of HOXA10,HOXA11,ER,PR and down-regulating expression of LIF.
作者
郝秀芳
王国华
和翔宇
唐磊
HAO Xiufang;WANG Guohua;HE Xiangyu;TANG Lei(The Third Affiliated Hospital of Beijing University of Chinese Medicine,Beijing 100029,China;Shunyi Hospital,Beijing Traditional Chinese Medicine Hospital,Beijing 101300,China)
出处
《现代中西医结合杂志》
2024年第24期3359-3364,3370,共7页
Modern Journal of Integrated Traditional Chinese and Western Medicine
基金
北京中医药大学青年教师项目(2019-JYB-JS-166)。
关键词
子宫内膜异位症
温肾化瘀
子宫内膜容受性
妊娠结局
同源框基因
endometriosis
warming kidney and dissipating blood stasis
endometrial receptivity
pregnancy outcome
homeobox gene
