补肾活血序贯法对薄型子宫内膜大鼠内膜形态及VEGF、Vimentin、CK12蛋白表达的影响

Effects of Sequential Method of Invigorating Kidney and Activating Blood on Endometrial Morphology and Expressions of VEGF,Vimentin and CK 12 Protein of Rats with Thin Endometrium

目的:观察补肾活血序贯法中药对薄型子宫内膜大鼠内膜形态、血管内皮生长因子(VEGF)蛋白、波形蛋白(Vimentin)和组织细胞角蛋白12(cytokeratin 12,CK12)表达的影响。方法:将SPF级雌性SD大鼠连续灌胃羟基脲8 d复制肾虚薄型子宫内膜模型,造模成功后50只大鼠随机分为模型组、中药低剂量组、中剂量组、高剂量组及补佳乐组(阳性对照组)每组10只,另取10只大鼠为假手术组。假手术组和模型组灌胃等量蒸馏水,补佳乐组灌胃补佳乐0.3 mg/kg。其余3组动情期分别灌胃给予低、中、高剂量的消癥合剂;动情前期、动情后期和动情间期分别灌胃给予低、中、高剂量的养膜助孕包,连续给药3个动情周期,共15 d。干预结束后,HE染色观察大鼠子宫内膜形态;免疫组化检测大鼠子宫内膜VEGF、Vimentin和CK12表达。结果:模型组大鼠子宫内膜厚度、腺体数和血管数明显低于假手术组(P<0.01)。中药中、高剂量组及补佳乐组大鼠子宫内膜厚度、腺体数和血管数明显高于模型组(P<0.05或0.01)。中药中、高剂量组和补佳乐组大鼠子宫内膜组织VEGF的表达量均明显多于模型组(P<0.01),中药高剂量组子宫内膜组织Vimentin蛋白表达量明显高于模型组(P<0.01),中药低、中、高剂量组和补佳乐组子宫内膜组织CK12的表达明显多于模型组(P<0.05或0.01)。结论:补肾活血序贯法中药可以增加肾虚薄型子宫内膜模型大鼠的子宫内膜厚度及内膜组织VEGF、Vimentin、CK12蛋白表达。

Objective:To observe the effects of the sequential method of invigorating kidney and activating blood on the morphology of endometrium and the expressions of endometrial VEGF,vimentin and CK 12 protein of rats with thin endometrium.Methods:50 female SD rats of SPF grade were randomly divided into model group,low-dose group,medium dose group,high-dose group,Progynova group(positive group)after 8 days of continuous administration of hydroxyurea with each 10 rats and 10 rats were taken as sham operation group.The sham operation group and model group were gavaged with 10 mL/kg distilled water,and the Progynova group was gavaged with 0.3 mg/kg Progynova,the three dose of Chinese medicine groups were given low,medium and high doses of Xiaozheng Mixture by gavage during estrus,low,medium and high doses of Yangmo Zhuyun Mixture were given by gavage in the proestrus stage,estrus stage and metestrus stage,respectively,for 3 estrus cycles,a total of 15 days.After the intervention,HE staining was used to observe the morphology of rats’endometrium;The expressions of vascular endothelial growth factor(VEGF),vimentin and cytokeratin 12(CK12)were detected by immunohistochemistry.Results:The endometrial thickness,the number of glands and blood vessels in the model group were significantly lower than those in the normal group(P<0.01).The endometrial thickness,the number of glands and blood vessels in the medium and high dose groups and Progynova groups were significantly higher than those in the model group(P<0.05 or 0.01).The expression of VEGF in the endometrium of the medium and high dose groups and the Progynova group was significantly higher than that of the model group(P<0.01).The expression of vimentin protein in endometrium of high dose group was significantly higher than that of model group(P<0.01).The expression of CK12 in endometrium of low,medium and high dose groups and Progynova group was significantly larger than that of model group(P<0.05 or 0.01).Conclusion:The sequential method of invigorating kidney and activating blood can increase the endometrial thickness and the expressions of VEGF,vimentin and CK12 protein in the endometrium of thin endometrium model rats.