| Objective Mongolian medicine mammary gland-Ⅰ(M-Ⅰ)has achieved satisfactory results in the treatment of hyperplasia of mammary glands(HMG).This study aims to explore the therapeutic effect and mechanism of Mongolian medicine Mammary Gland-Ⅰon mammary hyperplasia by regulating the expression of mitogen activated extracellular signal regulated kinase(MEK),extracellular signal regulated kinase(ERK)and proapoptotic protein cysteine containing aspartate-specific protease-3(Caspase-3)based on MEK / ERK pathway.Methods Among 36 SPF-grade SD healthy unpregnant female rats,30 rats were randomly selected to establish the mammary gland hyperplasia model by the sequential method of estrogen and progesterone,and 6 rats were treated as the normal control group(NC).After successful modeling,Thirty mammary gland hyperplasia model rats were randomly divided into model control group(MC),positive drug control group(PC),mammary gland-Ⅰlow dose group(M-ⅠL),mammary gland-Ⅰmiddle dose group(M-ⅠM),mammary gland-Ⅰhigh dose group(M-ⅠH)(n=6).After successful modeling,each group was given medicine for 4 weeks.M-ⅠL group was given M-Ⅰ0.5g /(kg·d),M-ⅠM group was given M-Ⅰ1 g /(kg·d),M-ⅠH group was given M-Ⅰ3 g /(kg·d),PC group was given tamoxifen citrate tablets 1.8 mg /(kg·d),NC group and MC group were given normal saline 0.5 ml / d.The body weight of rats and the diameter of the third pair of nipples under left axillary were recorded every week.At the end of the experiment,the third left breast tissue of rats was taken,and the pathological morphology of breast tissue was observed and graded by hematoxylin-eosin(HE)staining.Immunohistochemistry(IHC)and Western blot(WB)were used to detect the protein levels of MEK,ERK and Casepase-3 in the third right breast tissue of rats.Results The researchers observed the weight change of rats and found that there was no significant difference in the weight of rats between groups after the successful modeling(P>0.05).At the second week of drug administration intervention,M-ⅠH was significantly higher than that in MC group(P<0.01).At the third and fourth week,compared with NC group,the body weight of rats in MC group was significantly decreased(P<0.01).Compared with MC group,the body weight of rats in M-ⅠM and M-ⅠH groups was significantly increased,and was positively correlated with the dosage(P<0.01).Compared with M-ⅠL group,the body weight of rats in M-ⅠH group was significantly increased(P<0.01),which was closer to the normal group.The researchers observed the changes of rat nipple diameter and found that after the successful modeling,compared with the NC group,the nipple diameter of the model group was significantly widened(P<0.05).At the first week of intervention,compared with NC group,the diameter of MC group was still significantly widened(P<0.01),and compared with MC group,the diameter of M-ⅠH group was significantly decreased(P<0.01);at the second week,compared with NC group,the diameter of MC group was still significantly widened(P<0.01).Compared with MC group,the diameter of M-ⅠL,M-ⅠM and M-ⅠH groups was significantly decreased(P<0.01).Compared with PC group,the diameter of M-Ⅰ M and M-Ⅰ Hgroups was significantly decreased(P<0.01).Compared with M-IL group,the diameter of M-ⅠH group was significantly decreased(P<0.01).At the third and fourth week,compared with NC group,the diameter of MC group increased significantly(P<0.01).Compared with MC group,the diameter of PC,M-ⅠL,M-ⅠM and M-ⅠH groups decreased significantly(P<0.01).Compared with PC group,the diameter of M-ⅠL,M-ⅠM and M-ⅠH groups decreased significantly(P<0.01).Compared with M-ⅠL group,the diameter of M-Ⅰ H group decreased significantly(P<0.01).The researchers observed the pathological morphology of mammary gland under microscope and found that the number of mammary lobules in NC group was small and scattered.Acinar epithelial cells are mostly monolayer,acinar cavity is small,no secretions;in MC group,the structure of mammary gland was disordered,lobules and acinars of mammary gland were increased,multi-layer epithelium was visible,and some of them could be accompanied by papillary changes.Acinar expansion is obvious,a large number of secretions can be seen in the cavity;in the PC group,the fiber tissue was more in the lobule of breast,the number of acinar and ductal epithelial cells was 2-4 layers,and a little secretion was seen in the acinar cavity.M-Ⅰgroup with the increase of dose,M-ⅠL,M-ⅠM and M-ⅠH group acinar number decreased,epithelial cell layers decreased,acinar cavity narrowed,cavity secretions decreased,gradually tend to normal breast tissue.Researchers found after scoring,compared with NC group,the hyperplasia of mammary glands in MC group was severe(P<0.05),the hyperplasia of mammary glands in PC group was less than that in MC group(P<0.05),and the reduction in M-ⅠL,M-ⅠM and M-ⅠH groups was more significant(P<0.01).Compared with PC group,the hyperplasia of mammary gland in M-ⅠH group was significantly reduced(P<0.05).The researchers used immunohistochemical method to detect MEK,ERK and Caspase-3,and found that under the microscope,the positive staining particles of MEK,ERK and Caspase-3clustered or scattered in the cytoplasm of mammary gland were light brown staining particles.Compared with NC group,the expression of MEK and ERK in MC group was significantly increased(P<0.01).Compared with MC group,the expression of MEK and ERK in PC,M-Ⅰ L,M-ⅠM and M-ⅠH groups was significantly decreased(P<0.01).Compared with M-ⅠL and M-ⅠM groups,the expression of ERK in M-ⅠH group was significantly decreased(P<0.01).Compared with NC group,the expression of Caspase-3 in MC group was significantly decreased(P<0.01).Compared with MC group,the expression of Caspase-3 in PC,M-ⅠL,M-ⅠM and M-ⅠH groups was significantly increased(P < 0.01).Western blot was used to detect MEK,ERK and Caspase-3,and it was found that the expressions of MEK and ERK in PC,M-ⅠL,M-ⅠM and M-ⅠH groups were significantly lower than those in MC group(P<0.01).Compared with M-ⅠL group,the expression of MEK and ERK in M-ⅠH group was significantly decreased(P<0.01).Compared with the MC group,the expression of Caspase-3 in PC,M-Ⅰ L,M-Ⅰ M and M-Ⅰ H groups was significantly increased(P<0.01).Compared with M-ⅠL group,the expression of Caspase-3 in M-ⅠH group was significantly increased(P<0.01).Conclusion(1)Mongolian medicine Mammary I can significantly reduce the degree of mammary gland hyperplasia in rats and reduce the diameter of nipple.The effect of high dose Mammary gland-I is the most obvious,the earliest,better than tamoxifen.(2)High dose Mammary gland-I has the least effect on body weight of rats,the earliest effect and the best therapeutic effect.(3)In rat mammary gland tissues,MEK,ERK and Caspase-3 proteins were localized in cytoplasm.(4)Mongolian medicine Mammary-I can treat hyperplasia of mammary glands by inhibiting the expression of MEK / ERK cell signaling pathway and promoting the expression of pro-apoptotic protein Caspase-3,especially in high-dose M-I group.The mechanism of Mongolian medicine Mammary Gland-I in the treatment of HMG may be related to inhibiting the excessive proliferation of breast cells and promoting apoptosis. |
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