Study On Effect Of Rupifang By External Use In Treating Hyperplasia Of Mammary Gland Rat's Model With Stagnation Of Liver-Qi Type

Objective:To observe the effects of Rupifang on the levels of hormone in serum and pathological changes of Hyperplasia of Mammary Gland (HMG) rat's model with stagnation of liver-qi type and explore the mechanism of Rupifang by external use in preventing and treating HMG in order to offer scientific basis on the guidance to treat HMG effectively with Traditional Chinese Medicine in clinical.Method:Rat's model of HMG with stagnation of liver-qi type was made by combination of injecting benzestrofol and progesterone into the muscle and clamping the rat's tail. Meanwhile, low,middle and high doses of Rupifang was administrated by external use to treat HMG, with tamoxifen as a positive control. After 30 days, body weight, contour of breast and organ index were measured. Radio immunity method was used to detect the levels of sex hormone E2 and P in the serum. Mammary gland tissue was isolated, fixed in formalin and imbeded in paraffin, followed by HE staining to observe the pathological changes of mammary gland tissue. Immunohistochemistry staining was used to detect the expression level of C-erbB-2 and p16.Result:1. Reduction of the diameter and height of mammary papilla of the rat's HMG modle treating with different doses of Rupifang by external use was observed. More effect was found in the groups with higher doses of Rupifang than other groups (P＜0.01).2. The level of sex hormone E2 in the serum of rat's HMG modle with tagnation of liver-qi type was lower in the group treating with Rupifang by external use than other groups. More effect was found in the groups with low and high doses of Rupifang than other groups (P＜0.01).3. As for the pathological changes, improvement of hyperplasia level of mammary glands of the rat's HMG modle with Stagnation of Liver-Qi type were observed in the group treating with Rupifang than other groups including the group treating with tamoxifen, a standard therapeutic drug to treat HMG in clinic (P<0.01). More effect was found in the groups with middle and high doses of Rupifang than that with low doses (P<0.01).4. The expression level of C-erbB-2 was significantly lower in the groups treating with middle and high doses of Rupifang and tamoxifen compared with other groups (P<0.01). The expression level of C-erbB-2 was lower in the groups treating with middle and high doses of Rupifang than that with low doses (P<0.01).5. The expression level of p16 was significantly higher in the groups treating with different doses of Rupifang than that of other groups including the group treating with tamoxifen. The expression level of p16 in the groups of combination of diseases and symptom treating with middle doses of Rupifang was higher than that in the groups of simple deseases treating with Rupifang (P<0.05).Conclusion:Our data suggest that the external use of Rupifang is an effective method to prevent and treat HMG with stagnation of liver-qi type. It is supposed that Rupifang mayb play an important role in regulating the levels of hormone, improving the blood circulation, and finally preventing the hyperplasia of mammary glands effectively. Our data also show that Rupifang can decrease the expression level of C-erbB-2 and enhance the expression level of p16, which imlpy that Rupifang can influence the proliferation of the mammary glands and promote the cellular apoptosis to prevent or reverse the development of breast cancer originated from hyperplasia of mammary glands.