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目的:探究乌鸡白凤丸对衰老血虚大鼠子宫内膜VEGF及其受体表达的影响,并从分子生物学水平进一步探讨乌鸡白凤丸治疗肾虚血亏型月经不调的作用机理。方法:采用放血、饥饿加灌服羟基脲的方法制备肾虚血亏大鼠模型,并用乌鸡白凤丸进行干预。病理切片HE染色观察子宫内膜形态学结构变化;放射免疫法测定各组大鼠血清雌二醇、孕酮的表达水平;免疫组化检测子宫内膜VEGF、KDR及Flt-1蛋白的表达。结果:乌鸡白凤丸干预后能有效增加肾虚血亏大鼠子宫内膜的厚度;与空白组比较,模型组血清雌二醇水平明显降低(P<0.01)、孕酮水平明显增高,均具有显著统计学意义(P<0.01);子宫内膜VEGF蛋白、KDR蛋白、Flt-1蛋白表达减少(P<0.01);与模型组比较,乌鸡白凤丸高剂量组雌二醇水平增高,具有统计学意义(P<0.05);子宫内膜VEGF蛋白、KDR蛋白、Flt-1蛋白表达明显增加,具有显著统计学意义(P<0.01)。与模型组比较,乌鸡白凤丸低剂量组与中剂量组血清雌二醇、孕酮水平有升高的趋势,不具备统计学意义(P>0.05);子宫内膜子宫内膜VEGF蛋白、KDR蛋白、Flt-1蛋白表达明显增加,具有显著统计学意义(P<0.01)。结论:乌鸡白凤丸发挥作用的可能机制为通过升高血清雌二醇水平、降低孕酮水平来提高VEGF蛋白、KDR蛋白、Flt-1蛋白的表达,从而促进子宫内膜修复,达到调节月经的作用。
Objective: To investigate the effect of Wuji Baifeng Wan on endometrial VEGF and its receptor expression in aging blood deficiency rats, and to further explore the mechanism of Wuji Baifeng Wan in treating irregular menstruation caused by kidney deficiency and blood deficiency from the molecular biology level. Methods: The model of kidney deficiency and blood deficiency was prepared by blood-letting, starvation and irrigating with hydroxyurea. Intervention was made with Wuji Baifeng Wan. The morphological changes of endometrium were observed by HE staining. The expression of estradiol and progesterone in serum was detected by radioimmunoassay. The expressions of VEGF, KDR and Flt-1 protein in endometrium were detected by immunohistochemistry. Results: Wuji Baifeng Wan intervention could effectively increase the thickness of endometrium in rats with deficiency of kidney deficiency; compared with the blank group, the serum estradiol level was significantly lower (P <0.01) and the progesterone level was significantly higher in model group (P <0.01). The expression of VEGF protein, KDR protein and Flt-1 protein in endometrium were decreased (P <0.01). Compared with the model group, the estradiol level in high-dose group of Wuji Baifeng Wan increased (P <0.05). The expressions of VEGF protein, KDR protein and Flt-1 protein in endometrium were significantly increased (P <0.01). Compared with the model group, the serum estradiol and progesterone levels in low-dose group and middle-dose group of Wuji Baifeng Wan had no significant difference (P> 0.05); The expression of VEGF protein in endometrium, KDR protein and Flt-1 protein expression increased significantly, with significant statistical significance (P <0.01). Conclusion: The possible mechanism of Wuji Baifeng Wan is to improve the expression of VEGF, KDR and Flt-1 by increasing the level of serum estradiol and lowering the level of progesterone, so as to promote the repair of endometrium and regulate menstruation Role.