| Objectives:Hyperplasia of Mammary Gland, HMG,,also known as "breast cystic hyperplasia", is mammary epithelial hyperplasia,which is common and multiple in female in recent years and the incidence of which is rising too.2%of patients with breast hyperplasia have secondary cancer rates. However, the current treatment of breast hyperplasia is not very satisfactory.The experiment has been established to verify the feasibility of mesenchymal stem cells in the treatment of breast hyperplasia by intravenously injecting the rat model of mammary gland with bone marrow mesenchymal stem cells (bone marrow mesenchymal stemcells, BMSCs). The experiment uses flow cytometry and immunohistochemistry to detect β-catenin protein content and intracellular expression to explore whether mechanisms of mesenchymal stem cells for the treatment of breast hyperplasia is that dkk-1which is secreted by mesenchymal stem cells can be used to treat breast hyperplasia by inhibiting β-catenin protein expression in WNT pathway.Methods:The50SD rats were divided into five groups, including control group, model group, Rupixiao group, the tamoxifen group, BMSCs group,10in each group. The rats in Model group, Rupixiao group, tamoxifen group, BMSCs were injected with estradiol benzoate and progesterone to establish mammary hyperplasia model. After the establishment, the rats in both the control group and the model group were normally fed. Rats in the Rupixiao group were given Rupixiao orally once every day for28days, while the rats in Tamoxifen group were administered tamoxifen for28days. Rats in the Rupixiao group were given Rupixiao orally once every day for28days, while the rats in Tamoxifen group were administered tamoxifen for28days. The rats in the EMSCs group were injected with BMSCs via the tail vein in the1st,7th,14th, and21st day. Weekly measurements was implemented to detect the height and diameter of the second right nipples. After the experiment, blood was taken from the abdominal aorta to measure estrogen and progesterone levels,measuring the thymus, spleen, uterus, ovaries and other organs index.The methods of content flow cytometry and immunohistry were used to detect the content, the intracellular and extracellular expressions of β-catenin. HE slices are made to observe the morphology of the tissue cells.Results:1Nipple diameter variationThe nipple diameters of the rats in the control group and the model group changed slightly after therapy compared with those before therapy. The nipple diameters of the rats in the BMSCs group had the greatest changes and then came the Rupixiao group, followed by the tamoxifen group whose change was relatively small. Pearson correlation coefficient between the control group and that of other groups was statistically significant (P<0.05). Pearson correlation coefficient between the model group and other groups was statistically significant (P<0.05). And Pearson correlation coefficient between the BMSCs group and the Rupixiao group and the tamoxifen group was statistically significant (P<0.05). However, Pearson correlation coefficient between the Rupixiao group and the tamoxifen group was not statistically significant (P>0.05). Tip:BMSCs in the treatment of breast hyperplasia is more efficient than Rupixiao and tamoxifen. The effects of Rupixiao and tamoxifen in the treatment of breast hyperplasia has no significant difference.2Nipple height variationThe nipple height in the control group and the model group has changed a little, compared with that before therapy. The nipple height had the greatest change in BMSCs group after therapy, then came the Rupixiao group, followed by tamoxifen group whose change was relatively small. Pearson correlation coefficient between the control group and that of other groups was statistically significant (P<0.05). Pearson correlation coefficient between the model group and other groups was statistically significant (P<0.05). And Pearson correlation coefficient between the BMSCs group and the Rupixiao group and the tamoxifen group was statistically significant (P<0.05). However, Pearson correlation coefficient between the Rupixiao group and the tamoxifen group was not statistically significant (P>0.05). Tip:BMSCs in the treatment of breast hyperplasia is more efficient than Rupixiao and tamoxifen. The effects of Rupixiao and tamoxifen in the treatment of breast hyperplasia has no significant difference.3Hormone levelsEstrogen and progestin in each group was not statistically significant in Pearson correlation coefficient (P>0.05). The ratio between estrogen and progesterone was statistically significant (P<0.05). Tip:Hyperplasia may be caused by estrogen and progesterone hormone imbalance. Whether it is caused by estrogen imbalance alone or progesterone hormone imbalance alone needs further verification.4Organ index4.1Uterine IndexThe uterine index of the hyperplasia rats was smaller than that of the normal rats. Pearson correlation coefficient between the control group and that of other groups was statistically significant (P<.05). Pearson correlation coefficient between the model group and other groups was statistically significant (P<0.05). And Pearson correlation coefficient between the BMSCs group and the Rupixiao group and the tamoxifen group was statistically significant (.P<0.05). Pearson correlation coefficient between the Rupixiao group and the tamoxifen group was statistically significant (P<0.05). The weight of uterus in rats with hyperplasia was relatively small compared with that of normal rats.4.2Ovarian indexThe ovarian index of the hyperplasia rats was smaller than that of the normal rats. Pearson correlation coefficient between the control group and that of other groups was statistically significant (P<0.05). Pearson correlation coefficient between the model group and the BMSCs group was statistically significant (P<0.05). Tip:The weight of ovary in hyperplasia rats was relatively small, compared with that of normal rats.4.3Thymus indexThere was no statistical difference between the groups (P<0.05). Tip: Hyperplasia may have no effect on the thymus.4.4Adrenal IndexPearson correlation coefficient among normal group, Rupixiao group, and tamoxifen group was statistically significant (P<0.05).5β-catenin expressionThe rats in the control group had the lowest β-catenin level. The P-catenin level of the rats in BMSCs group and model group was high, while the β-catenin level of the rats in Rupixiao group and tamoxifen group was relatively low. Pearson correlation coefficient between the BMSCs group and tamoxifen group or control group was statistically significant (P<0.05). Pearson correlation coefficient between BMSCs group and model group or the Rupixiao group was not statistically significant (P>0.05). Pearson correlation coefficient between the Rupixiao group and tamoxifen group was not statistically significant (P>0.05). Tip:Mesenchymal stem cells did not affect the expression of β-catenin protein in breast tissue.6Immunohistochemistry case of p-cateninIn the control group, P-catenin was cell membrane staining, while in the model group it was plasma and nuclear staining. β-catenin in both Rupixiao group and was tamoxifen group cytoplasmic staining. β-catenin in BMSCs was plasma and nuclear staining. The staining degree in model group was similar to that in BMSCs group, and the staining degree in Rupixiao group was similar to that in tamoxifen group. Tip:Mesenchymal stem cells did not affect the expression of β-catenin protein in breast tissue.7Tissue cell morphology observation under microscopeBMSCs treatment reduced the breast hyperplasia syndrome in the breast tissue, the degeneration of epithelial cells disappeared, and most of tissue structures returned to regular ones. Hyperplasia of the breast tissue became slight by applying Rupixiao and tamoxifen, and the tissue structure had been restored to some extent.Conclusions:1Mesenchymal stem cell is more effective than tamoxifen and Rupixiao in the treatment of breast hyperplasia. Mesenchymal stem cells have the potential to become a new strategy for the treatment of breast hyperplasia.2This experiment suggests that mesenchymal stem cells may regulate estrogen, progesterone levels to treat breast hyperplasia. Whether breast hyperplasia is caused by estrogen imbalance alone or progesterone hormone imbalance alone needs further verification.3Mesenchymal stem cells do not affect the expression of β-catenin protein in breast tissue. More experiment are needed to done to illustrate that the breast hyperplasia treatment is achieved by prohibiting WNT pathway. |
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