| Objects:The suitable dose used for external application of rupifang in mammary gland hyperplasia rats model was ascertained from the previous study. To discuss the mechanism of external application of rupifang to mammary gland hyperplasia. And a determinate theoretic basis for clinical prevention and treatment of HMG of traditional Chinese medicine were disscussed for the development and application of new drugs.Methods:30 female adult SPF SD rats were divided into 5 groups randomly, which were normal control group, model group, the sanjierupi group, the tamoxifen group, the rupifang group, with 6 rats in each group. Mammary gland hyperplasia model were established in groups excepted the normal control group, and different of external application drugs were used in each group. Detection were taken for the Measurement of rats'weight, the change of the breasts, the organ coefficient of uterus, liver and spleen, the serum of estradiol and progesterone levels by radioimmunoassay, and the serum SOD activity and MDA level by colorimetric method, the pathology changes and the expression of ER of the breast tissue by immunohistochemical method.Results:1. The rat body weight changes:The rats were weighed every weekend, by repeated measures analysis of variance it showed that:time effect was statistically significant (F= 218.218, P=0.000), time and treatment interaction effect also was significant (with suitable for small samples Huynh-Feldt method, F=2.091, P=0.024).2. The rat nipple diameter and height changes:The first 2,3 nipple diameter and height of the model rats increased significantly compared with the control group (P<0.01); after drugs intervention, each group of nipple height, diameter was less than the model rats (P <0.01 or P<0.05).3. Organ coefficient:The organ coefficient of uterus and liver of model group were increased compared with the normal control group (P<0.01), the organ coefficient of spleen was not significant. After drugs intervention, the rat uterus coefficient reduced (P<0.01), the organ coefficient of liver was significantly increased compared with normal control group (P <0.01), but without statistic difference (P> 0.05), spleen coefficient of each group had no significant difference between the groups.4. Changes of serum sex hormones:The serum estradiol in model rats were significantly higher, but lower in serum progesterone compared with normal control group (P<0.01). After drugs intervention, except sanjierupi group, the other groups were significantly lower serum estradiol and higher serum progesterone (P<0.01). 5. Changes of serum SOD activity and MDA content:Compared with normal control group, serum SOD activity in model group decreased and the serum MDA level was higher significantly (P<0.01). After drugs intervention, except sanjierupi group the other groups had significant effect on SOD activity decreased, the rest of the modeling intervention groups significantly increased the role of SOD activity (P<0.01), After drugs intervention the serum MDA levels in groups of sanjierupi group, tamoxifen group and rupifang group were significantly lower(P<0.01 or P<0.05).6. Histopathology changes:Compared with the normal control group, all the rat breast pathological tissues of model were severe hyperplasia, (P<0.01); after drugs intervention, the breast tissue histopathology of each group of rats were improved significantly (P<0.01 or P <0.05).7. ER expression changes:The normal control group in the mammary gland expression of ER negative, while the other groups in the positive expression of ER compared with normal control group were significantly higher (P<0.01); the tamoxifen group and the rupifang group had a better effect than the group of sanjierupi(P<0.05). The rat mammary gland by tumor grade and statistics suggests that:With the severity of breast tissue proliferation, ER expression positive ratio increased, the pathological grade and ER expression is closely related (Linear trend test, the total variance x2=94.031, v=6, P=0.000; Linear regression component x2=71.664, v=1, P=0.000, Deviate from the linear regression component x2=22.367, v=5, P<0.005; rank correlation coefficient=0.808, P=0.000).Conclusions:The external application of rupifang could largely reduce the degree of hyperplasia of the HMG rats and reduce the nipple diameter and height, lower the organ coefficient of uterus effectively, lower serum estradiol and increased serum progesterone, increase the serum SOD activity, lower serum MDA, improve the proliferation of pathological morphology, inhibition of ER expression in breast tissue of them, effectively control mammary glands of the HMG rats. The mechanism might be regulate the serum hormone levels of the rats of mammary gland hyperplasia by reducing free radical, oxygen free radical damage and lipid peroxidation, inhibiting the expression of hormone receptors, resulting in the role of inhibition and treatment of the HMG.
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