Zheng Juan, Tan Yong
Henan Provincial Hospital of Traditional Chinese Medicine, Zhengzhou 450002, China.
Nanjing University of Chinese Medicine, Nanjing 210046, China.
Sichuan Da Xue Xue Bao Yi Xue Ban. 2021 Mar;52(2):235-240. doi: 10.12182/20210160508.
To explore the influence for combination of Dingkun Dan with estradiol valerateon in treating rats with thin endometrium with Kidney-Yang deficiency based on Wnt/β-catenin signaling pathway.
The estrous period 40 rats were randomly divided into the normal control group, the model group, the Dingkun Dan group, the estradiol valerateon group and the combination group, with 8 rats in each group. In addition to the normal control group, the rat model of thin endometrium with Kidney-Yang deficiency was established in other groups. The control group used free diet, the model group was given distilled water, the estradiol valerateon group was treated with progynova by gavage at 0.3 mg/(kg·d) , Dingkun Dan group was treated with Dingkun Dan by gavage at 2.26 g/(kg·d), and the combined group was given Dingkun Dan at 2.26 g/(kg·d) on the basis of progynova at 0.3 mg/(kg·d). After 3 estrous cycles, the rats were killed and harvested. HE staining was used to observe histopathologic changes in endometrium. The expression of VEGF in rats endometrium were detected by immunohistochemistry. The expression of β-catenin, E-cadherin and MMP-9 protein in rat endometrium was detected by Western blot.
Compared with the control group, the uterine cavity was narrowed or enlarged, the endometrium glands and blood vessels were sparse, and the endometrium was thinner significantly in the model group ( <0.01); the levels of VEGF was decreased significantly ( <0.01), while β-catenin, E-cadherin and MMP-9 were increased significantly ( <0.01). Compared with the model group, more endometrial glands, rich intimal vessels, the endometrium were thickened significantly in the 3 treatment groups ( <0.01 or <0.05); the levels of VEGF was increased differently. The protein levels of β-catenin and E-cadherin were significantly decreased in each treatment group ( <0.01), and MMP-9 were significantly decreased in the Dingkun Dan group and in the combination group ( <0.01). Compared with the estradiol valerateon group, the level of β-catenin in Dingkun Dan group was higher, and MMP-9 was lower ( <0.05 or <0.01). The levels of β-catenin and MMP-9 in the combination group were significantly decreased ( <0.01). Compared with the combination group, the levels of β-catenin was increased significantly, while decreased signicantly in Dingkun Dan group ( <0.01 or <0.05).
Dingkun Dan combined with estradiol valerateon can increase the thicken of the endometrium by up-regulation of VEGF, while down-regulate of β-catenin, E-cadherin and MMP-9 in rats with Shen-Yang deficiency and thin endometrium.
基于Wnt/β-连环蛋白信号通路探讨定坤丹联合戊酸雌二醇对肾阳虚型薄型子宫内膜大鼠的影响。
将40只处于发情期的大鼠随机分为正常对照组、模型组、定坤丹组、戊酸雌二醇组和联合组,每组8只。除正常对照组外,其余各组建立肾阳虚型薄型子宫内膜大鼠模型。对照组自由饮食,模型组给予蒸馏水,戊酸雌二醇组按0.3 mg/(kg·d) 灌胃给予补佳乐,定坤丹组按2.26 g/(kg·d) 灌胃给予定坤丹,联合组在按0.3 mg/(kg·d) 给予补佳乐的基础上按2.26 g/(kg·d) 给予定坤丹。3个发情周期后处死大鼠并取材。采用HE染色观察子宫内膜组织病理学变化。采用免疫组化法检测大鼠子宫内膜VEGF表达。采用蛋白质印迹法检测大鼠子宫内膜β-连环蛋白、E-钙黏蛋白和基质金属蛋白酶-9蛋白表达。
与对照组比较,模型组子宫腔狭窄或增大,子宫内膜腺体和血管稀疏,子宫内膜明显变薄(P<0.01);VEGF水平明显降低(P<0.01),而β-连环蛋白、E-钙黏蛋白和基质金属蛋白酶-9明显升高(P<0.01)。与模型组比较,3个治疗组子宫内膜腺体增多,内膜血管丰富,子宫内膜明显增厚(P<0.01或P<0.05);VEGF水平不同程度升高。各治疗组β-连环蛋白和E-钙黏蛋白蛋白水平明显降低(P<0.01),定坤丹组和联合组基质金属蛋白酶-9明显降低(P<0.01)。与戊酸雌二醇组比较,定坤丹组β-连环蛋白水平较高,基质金属蛋白酶-9较低(P<0.05或P<0.01)。联合组β-连环蛋白和基质金属蛋白酶-9水平明显降低(P<0.01)。与联合组比较,定坤丹组β-连环蛋白水平明显升高,而基质金属蛋白酶-9明显降低(P<0.01或P<0.05)。
定坤丹联合戊酸雌二醇可上调肾阳虚型薄型子宫内膜大鼠VEGF表达以增加子宫内膜厚度,同时下调β-连环蛋白、E-钙黏蛋白和基质金属蛋白酶-9表达。