左、右归丸对控制性超排卵小鼠子宫内膜VEGF及Ang影响的研究

Effect of Zuogui Wan/Yougui Wan on VEGF and Ang in endometrial of COH mice

目的:基于VEGF及Ang探讨补肾复方左、右归丸对控制性超排卵小鼠子宫内膜容受性的影响。方法:采用GnRH-a+PMSG+HCG建立控制性超排卵小鼠模型,将其分为模型组、左归丸组、右归丸组、阿司匹林对照组、同时设正常对照组。在造模的同时,左、右归丸组分别按6、3、1.5g/kg左、右归丸灌胃阿司匹林对照组按0.2g/kg阿司匹林灌胃,模型组、正常对照组以等体积生理盐水灌胃,每天1次,连用11天。采用免疫组化法对子宫进行病理切片制作,通过IPP7.0图像分析系统检测子宫内膜VEGF、VEGF-R、Ang-1、Ang-2及Tie-2蛋白表达水平,采用荧光定量PCR技术检测阴道组织VEGF、VEGF-R、Ang-1、Ang-2及Tie-2 mRNA表达水平。结果:正常对照组VEGF/VEGF-R蛋白表达、VEGF-R mRNA表达、Ang/Tie蛋白表达和Ang-1 mRNA表达均明显高于模型组,与模型组比较,右归丸3g/kg剂量组及6g/kg剂量组VEGF蛋白表达明显增高;右归丸3g/kg剂量组VEGF、VEGF-1 mRNA表达(2.02±0.92、2.10±0.39),6g/kg剂量组VEGF、VEGF-1 mRNA表达(1.52±0.33、1.89±0.22)明显升高;右归丸3g/kg剂量组及6g/kg剂量组Ang-1、Ang-2蛋白表达明显增高,左、右归丸1.5g/kg剂量组、3g/kg剂量组、6g/kg剂量组tie-2明显增高;右归丸1.5g/kg剂量组Ang-1、Ang-2、Tie-2 m RNA表达(3.52±0.98、1.84±0.23、2.68±0.82)升高;3g/kg剂量组Ang-1、Ang-2、Tie-2 m RNA表达(3.48±0.89、1.88±0.26、2.69±0.71)、6g/kg剂量组Ang-1、Ang-2、Tie-2mRNA表达(3.57±0.98、1.96±0.32、2.64±0.66)明显升高。结论:补肾复方左、右归丸可通过调节控制性超排卵小鼠子宫内膜VEGF/VEGF-R及Ang/Tie蛋白和mRNA表达而改善子宫内膜容受性,其中补肾阳的右归丸比补肾阴的左归丸效果明显。

Objective： To explore the mechanism of Zuo Gui Wan/You Gui Wan in improving endometrial receptivity in COH mice by VEGF and Ang. Methods： COH mice model was established by using GnRHa ＋ PMSG ＋ HCG,and they were divided them into 8 groups i. e. the model group,Zuo Gui Wan/You Gui Wan groups,aspirin group,in addition the control group was set. During mice model was made,mice in Zuo Gui Wan/You Gui Wan（6 g,3 g,1 g/kg） groups,0. 2 g/kg aspirin group and,the model group（saline） and control group（saline）were administered（ig）,respectively,once a day for 11 days. Uterine pathological sections were observed by immunohistochemical method,endometrial VEGF,VEGF-R Ang-1,Ang-2 and Tie-2 protein,and protein expression level were detected by IPP7. 0 image analysis system,endometrial VEGF,VEGF-R Ang-1,Ang-2 and Tie-2 protein mRNA expression level were detected by fluorescence quantitative PCR technique. Results： The expression of VEGF/VEGF-R protein,expressions of VEGF-R and mRNA,expressions of Ang/Tie protein and expressions of Ang-1 and mRNA in the control group were higher than those in the model group（P〈0. 01）. Compared with the model group,expressions of VEGF protein and mRNA（2. 02 ± 0. 92,2. 10 ± 0. 39） in 3 g/kg You Gui Wan group,（1. 52 ± 0. 33、1. 89 ± 0. 22） in 6 g/kg group was significantly increased（P〈0. 05）; Tie-2 in Zuo Gui Wan/You Gui Wan（1. 5 g,3 g,6 g/kg） groups significantly increased（P〈0.05）; Ang-1,Ang-2 and tie-2 mRNA expression（3. 52 ± 0. 98,1. 84 ± 0. 23,2. 68 ± 0. 82） in 1. 5 g/kg You Gui Wan group,（3. 48 ± 0. 89,1.88 ± 0. 26,2. 69 ± 0. 71） in 3 g/kg group,（3. 57 ± 0. 98,1. 96 ± 0. 32,2. 64 ± 0. 66） in 6 g/kg group were significantly increased（P〈0. 05）.Conclusion： Zuo Gui Wan/You Gui Wan improve endometrial receptivity through regulating expressions of VEGF/VEGF-R and Ang/Tie protein and mRNA in the endometrium of COH mice.