| Radix Codonopsis Lanceolata is a kind of common herbal medicine used by the people.A large number of traditional Chinese medicine records indicate that the Radix Codonopsis Lanceolata has the effect of stimulate the menstrual flow and treating postpartum lactation.Polysaccharide is one of the most important biological activities and has been extensively studied.And a large number of clinical data show that obesity reduces the breastfeeding success rate of women and the quality of breast milk during lactation.However,the mechanisms involved and better preventive treatment options are not fully understood.Based on this,this paper uses the Radix Codonopsis Lanceolata polysaccharide(PCL)as experimental material,mainly to study for 6 weeks of puberty,10 weeks of maturity,7-9days of pregnancy,12-14 days of pregnancy,day 1 of lactation and day 7 of lactation.During these six periods,PCL was used to suppress phenomena caused by HFD,including slow duct development,reduced breast cell proliferation and differentiation,postpartum hypoglycemia,and the decline in the quality of milk ingredients.The experimental results of this paper are mainly:(1)Long-term HFD do affect the growth of mammary ducts in mice,including excessive accumulation of mammary fat pads,and uneven distribution and limited development of mammary ducts.The results of the whole body carmine staining showed that PCL can significantly improve the phenomenon caused by the HFD,including the decrease of mice mammary duct density,branching and uneven distribution.In addition,HE staining results further indicate that PCL can improve the reduction of the thickness of the mammary duct epithelial layer in mice due to the HFD.These results indicate that PCL can significantly improve the phenomenon caused by HFD,including decreased ductal duct density,uneven distribution,and limited development.(2)PCL can improve the symptoms of decreased proliferation and differentiation of mice mammary ductal epithelial cells induced by HFD,so as to promote the development of mice mammary gland.The results of immunohistochemistry by α-SMA and KI67 showed that PCL can improve the linear encapsulation integrity of mammary gland epithelial cells and endothelium in model group and the degree of breast cell proliferation.The results of the flow cytometry showed that compared with the LFD group mice,the HFD caused abnormal changes in the ratio of basal cells and luminal cells in the breast,and the increase of luminal progenitor cells.However,PCL can significantly ameliorate theproportion of basal cells and luminal cells in the breast,and reduce the number of luminal progenitor cells to promote the developmental maturation of the luminal cells.At the same time,the results of β-casein immunohistochemistry further proved that the model group was delayed in the pre-lactation stage and the degree of differentiation was lower in the late lactation stage,while PCL could significantly increase the differentiation of the luminal cells in the pre-lactation and late lactation stages.These results indicate that PCL can significantly improve the symptoms of decreased proliferation and differentiation of ductal epithelial cells caused by HFD,and promote the differentiation and maturation of luminal cells,thereby promoting the process of lactation in mice.(3)PCL can improve a series of phenomena induced by HFD,including decreased postpartum lactation,decreased in the quality of milk ingredients,slow growth of postpartum pups and low survival of offspring.By counting the lactation yield,the weight change of postpartum pups and the survival rate of the offspring,it was found that the postpartum lactation yield of mice induced by HFD was significantly lower than that of LFD group and PCL group.Moreover,the weight of the postpartum pups in HFD group was much lower than that of the LFD group and the PCL group.At the same time,the HFD group caused a certain degree of mortality in postpartum pups.In addition,the content of triglyceride,lactose and casein,the main ingredients of postpartum milk,was analyzed.The triglyceride,lactose and casein of the model group were found to be lower than the LFD group and the PCL group.These results indicate that PCL can improve a series of phenomena induced by HFD,including decreased postpartum lactation,decreased in the quality of milk ingredients,slow growth of postpartum pups and low survival of offspring.(4)Postpartum hypogalactia induced by HFD can be improved by PCL intervention in the relevant signaling pathways of postpartum lactation regulation.A high-fat diet can cause a decrease in the amount of prolactin in the blood of mice,which in turn affects the gene expression and activity changes of a series of key proteins downstream,and ultimately leads to postpartum hypogalactia.The results of this experiment showed that in the comparison model group,PCL was found to increase the content of prolactin in serum.Furthermore,by Western-blot analysis,it was found that PCL can increase the protein expression levels of pJAK2(phosphorylated JAK2),PRLR(prolactin receptor)and p63 in mouse mammary gland,and decrease the protein expression level of Nrg1.In addition,immunohistochemistry results showed that PCL could increase the expression of pSTAT5(phosphorylated STAT5).PCR results showed that the mRNA expression levels of ERBB4,p63 and Gata-3 in the PCL group were higher than those in the model group,while the PCL group of the mRNA expression of Nrg1 was lower than that of the model group.Theseresults indicate that for HFD-induced postpartum hypogalactia model,PCL can promote the proliferation,differentiation and maturation of breast cells to achieve the final secretion of milk by interfering with the relevant signaling pathways of postpartum lactation regulation.The above results indicate that PCL can improve the proliferation of mammary gland cells and the differentiation and maturation of luminal cells by reducing the accumulation of mammary fat pad in mice induced by HFD and finally achieve the goal of improving postpartum hypogalactia. |
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