| Objective:Modern medicine holds that there is a certain correlation between breast hyperplasia and canceration,The syndrome of liver depression is one of the important TCM syndromes of breast hyperplasia.Clinical and experimental studies have shown that Shugan Jieyu Formula-Chaihu Shugan Powder has a certain curative effect on the prevention and treatment of breast hyperplasia.This article suggests that there may be some kind of correlation between liver depression syndrome,breast hyperplasia,and breast cancer.Therefore,in the replication of syndrome models,We should observe changes in the gonad axis and mammary glands of the rat model and explore the expression of mammary gland hyperplasia and related precancerous genes at the molecular level.In addition,based on the theory of correlation between prescription and syndrome,the regulating effect of Chaihu Shugan Powder on the symptoms of hepatic depression and gonadal axis in the model rats and the mechanism of prevention and treatment of mammary gland hyperplasia were observed.The paper includes two parts:literature review and experimental research.The literature review focuses on the collection and management of literature on the prevention and treatment of breast hyperplasia and the relationship between breast hyperplasia and breast cancer.Experimental study explores the TCM syndromes of female rats under this model,the Abnormal Mechanism of Precancerous Lesion of Breast Cancer and the Correlative Effects of Chaihu Shugan Powder.Methods:Rats were randomly divided into normal control group,liver depression syndrome(model group),Chaihu Shugan Powder High Dose Group(H group)and Chaihu Shugan Powder Low Dose Group(L group).The other three groups were consecutively established by chronic restraint stress for 4 weeks,except the normal group.After modeling for 14 days,the rats in the H group and L group were given the Chaihu Shugan Powder for the dose of 1.24g/kg and 0.3 1g/kg(About 2 times and 0.5 times adult dose)once a day for 2 weeks,the rats in Normal groups and model groups were given equal amounts of distilled water.The vaginal smears of each group were prepared on the 27th day of the experiment to observe the estrus cycle,and the rats in the pre-estrus stage/moral stage(28-30 days of experiment)were selected for anesthesia to kill.Blood was taken from the abdominal aorta to separate serum and plasma;hypothalamus and mammary gland tissue were removed at low temperature for detection.Observing indicators:(1)Liver depression related indicators:1)General appearance:During the modeling period,the changes of appearance behavior and body weight of each group of rats were observed and recorded per week.Sugar water consumption test was performed at the end of the 2nd and the 4th weekend,and the open field test was performed at the 4th weekend.2)Liver depression related laboratory indicators:CRH,ACTH,CORT,5-HT,DA,E and NE.(2)Sex hormones and their axis of regulation:GnRH,FSH,LH,PRL,P,T,E2.(3)Breast organ:Breast histopathological observation.(4)Pre-cancerous genetic indicators:TFF1,TFF3,TIMP-2 and Survivin.(5)PRL-mediated signal pathway related indicators:PRLR,AKT,BAD,Bcl-2,MDM2 and P53.All data were expressed as x±S,and one-way ANOVA was used to test the comparison between groups using the Student Newman Keuls Test.All data are statistically processed using SPSS 20.0 for windows software.All charts were made with GraphPad Prism 7 software.Results:(1)Syndrome indicators:1)General appearance:Compared with the normal group,the behavioral scores of the rats in the model group increased significantly from week 2 to week 4(p<0.01),Obvious decrease in body weight(p<0.01);The model rat's SPT was significantly increased at the 2nd week,and significantly decreased at the 4th week(p<0.01),The stay time,the number of crawls,and the climbing mileage of the central area of the open field test were all significantly reduced at the 4th week(p<0.01).Compared with the model group,the behavior scores of the rats in the H group were significantly decreased in the 3rd to 4th week(p<0.01),and the body weight was significantly increased in the 4th week(p<0.01).The SPT in the H group was significantly increased in the 4th week(p<0.01),The stay time,number of crawls,and number of climbs in the central area of the open field test in the H group were significantly higher(p<0.01).There was no significant change in the above indicators in the L group.2)Changes in laboratory indicators:Compared with the normal group,The levels of CRH in the hypothalamus,ACTH,CORT and E in the blood increased significantly in model rats(p<0.01 or p<0.05),Blood levels of 5-HT,NE,and DA show significant reductions(p<0.01 or p<0.05).Compared with the model group,The levels of CRH in the hypothalamus,ACTH,CORT in the blood decreased significantly in H group rats(p<0.01 or p<0.05)5 Blood levels of 5-HT and NE increased significant(p<0.01 or p<0.05),Blood levels of ACTH decreased significantly in L group rats(p<0.05),Blood levels of 5-HT and DA increased significant(p<0.01).Compared with the H group,The levels of CRH in the hypothalamus,DA in the blood increased significantly in L group rats(p<0.01 or p<0.05),Blood levels of 5-HT decreased significant(p<0.01).(2)Changes in sex hormones and hormones:Compared with the normal group,The levels of GnRH in the hypothalamus,FSH,LH,PRL,T,E2,P in serum increased significant in the model group(p<0.01).Compared with the model group,The levels of GnRH in the hypothalamus,LH,PRL,T,P in serum decreased significant in the H group(p<0.01 or p<0.05),The serum levels of LH,PRL,T,E2 decreased significant in the L group(p<0.01 or p<0.05).Compared with the H group,The levels of GnRH in the hypothalamus increased significant in the L group(p<0.01);There was no significant difference in E2/P between the four groups.(3)Breast changes:In the normal group,the acinar shape of the breast was regular,there was no expansion of the lumen of the gland,and no secretions were seen in the acinar and duct.Compared with the normal group,The acinar shape of the mammary gland was irregular in the model group,the lumen of the glandular duct was dilated,and acinar and ductal secretions increased.Compared with the model group,The shape of the acinar of the mammary glands of rats in the H group was regular,but it was still accompanied by irregular acini,with few acini and endocrine secretions,mammary glands see rules,But most acini are irregular in rats of L group.(4)Changes in pre-breast cancer genes:Compared with the normal group,The levels of Survivin and TIMP-2 in mammary glands of model rats were significantly increased(p<0.01),The serum levels of TFF1 and TFF3 decreased significantly(p<0.05).Compared with the model group,The levels of Survivin and TIMP-2 in mammary glands of rats in H group were significantly decreased(p<0.01),The levels of TIMP-2 in mammary glands of rats in L group were significantly decreased(p<0.01).Compared with the H group,The levels of Survivin in mammary glands of rats in L group were significantly increased(p<0.01).(5)Change of PRL-mediated signal pathway related indicators:Compared with the normal group,The levels of PRLR,AKT,BAD and Bcl-2 in mammary glands of model rats were significantly increased(p<0.01 or p<0.05),There was no significant difference in MDM2 and P53.Compared with the model group,The levels of PRLR,AKT and BAD in mammary glands of rats both in H and L group were significantly decreased(p<0.01),The levels of Bcl-2 in mammary glands of rats in L group were significantly decreased(p<0.05).Conclusions:The female rats with liver depression syndrome model reproduced by the chronic binding method showed changes in appearance behaviors and laboratory indicators related to TCM syndromes.In addition,it is accompanied by hyperactivity of the gonadal axis and pathological hyperplasia of the breast.Breast tissue associated with pre-cancerous gene expression and abnormalities of PRL-related signaling pathways,suggesting that the molecular mechanism of precancerous lesions of mammary gland hyperplasia of liver depression syndrome.Chaihu Shugan Powder not only has a good effect on improving the TCM syndromes and laboratory indexes of female rats with liver depression syndrome,but also has certain preventive and therapeutic effects on the abnormality of gonadal axis and mammary gland hyperplasia in model rats.The formula also modulates breast cancer/suppressor gene expression in rat mammary glands while improving mammary gland hyperplasia and serum PRL in model rats,involving regulation of PRL-related signaling pathways.The results of this study provide some biological evidence for the speculation of the Chinese medicine "Liver Depression Syndromes--serum level of PRL ?--mammary hyperplasia--aberrant expression of related oncogenes--regulatory effect of Chaihu Shugan Powder".This study provides the basis understanding of the scientific connotations of the association between traditional Chinese medicine "Liver depression syndrome" and female breast glands,and the TCM syndrome differentiation and treatment of "prevention and control before getting sick". |
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