Discuss Mechanism Of Gastrointestinal Dysfunction In Liver-stagnation Pattern Of Perimenopausal Rats From MuYu KeTu

ObjectiveFrom the perspective of intestinal flora,gastrointestinal hormones and gastrointestinal motility to discuss the mechanism of gastrointestinal dysfunction in liver-stagnation of perimenopausal rats,in order to enrich the theoretical connotation of MuYu KeTu.Methods1.2-month-old of forty-eight SPF-grade female SD rats were randomly divided into sham operation group?sham group?,perimenopausal group?PMP group?,liver-stagnation pattern in perimenopause group?model group?and Chaihu Shugan San group?CSS group?according to body mass of rats after 1 month of adaptive feeding.Adipose tissue near the ovaries was removed from the sham group,and the ovaries was removed from the other three groups.Liver-stagnation pattern of model group and CSS group were established by Chronic Unpredictable Mild Stress?CUMS?.2.Smears of the vaginal exfoliated cells of rats were observed by Hematoxylin-eosin Staining?HE?.3.Sham group,PMP group and model group were given saline,while CSS group was given Chaihu Shugan San once a day for 21 days consecutively from the first day of CUMS.4.Body mass,feed consumption,total moving distance,vertical times,central area time,forced swimming immobility time and 1%sugar water consumption rate of rats was observed.5.Gastrointestinal motility of rats were detected by gastric emptying and intestinal propulsion rate.6.Serum estradiol?E₂?,5-hydroxytryptamine?5-HT?,substance P?SP?and motilin?MTL?of rats were detected by enzyme-linked immunosorbent assay?ELISA?.7.Intestinal flora in faeces of rats were detected by 16s rDNA amplification sequencing.8.Colon tissues mRNA of Occludin and Zona Occludens 1?ZO-1?of rats were detected by real-time fluorescence quantitative PCR?q-PCR?.9.Colon tissues protein of Occludin and ZO-1 of rats were detected by Western Blot?WB?.10.Serum interleukin-1??IL-1??,interleukin-6?IL-6?and tumour necrosis factor??TNF-??of rats were detected by suspension microarray.Results1.The exfoliated cells in the vagina of sham group was composed of epithelial cells,keratinocytes,mixture of inclusions keratinocytes and epithelial cell,and leukocytes.The other three groups of vaginal exfoliated cells present continuous white blood cells,which means lost the regular estrus cycle,and continue in the interestrus period.2.In terms of open-field experiment of rats,The total movement distance and vertical times of model group was decreased before and after modeling?P<0.01?.In terms of forced immobility time of rats,the model group was higher than sham group?P<0.05?.CSS group was decreased compared with model group?P<0.01?.In terms of 1%sugar water consumption rate of rats,model group was decreased compared with PMP group?P<0.05?.CSS group was increased compared with model group?P<0.01?.3.In terms of body mass of rats,PMP group was heavier than sham group?P<0.01?,and model group was lighter than PMP group?P<0.01?.In the terms of weight growth rate of rats,PMP group increased compared with sham group?P<0.01?,model group was decreased compared with PMP group?P<0.01?,and CSS group was increased compared with model group?P<0.05?.In terms of feed consumption of rats,there was a decreasing trend in model group compared with PMP group,and an increasing trend in CSS group compared with model group.4.In terms of serum E₂ of rats,PMP group,model group and CSS group were decreased compared with sham group?P<0.01?.However,there was no significant difference between PMP group,model group and CSS group.In terms of serum 5-HT of rats,model group was decreased compared with PMP group?P<0.01?,CSS group was increased compared with model group?P<0.01?.5.In terms of gastric residual rate of rats,model group was higher than PMP group?P<0.01?,CSS group was lower than model group?P<0.01?.In terms of intestinal propulsion rate of rats,model group was lower than PMP group?P<0.01?,CSS group was higher than model group?P<0.01?.6.In terms of serum MTL of rats,model group was decreased compared with sham group?P<0.01?,CSS group was increased compared with model group?P<0.01?.In terms of serum SP of rats,PMP group was decreased compared with sham group?P<0.05?,model group was decreased compared with PMP group?P<0.01?,CSS group was increased compared with model group?P<0.01?.7.In terms of MTL and SP of rats'small intestine,model group was decreased compared with PMP group?P<0.01?,CSS group was increased compared with model group?P<0.01?.8.In terms of rats'intestinal flora,species diversity was changed in each group,and there was a significant difference between each groups?P<0.01?.There was a trend of decrease in Lactobacillus of rats'intestinal flora between sham group and PMP group.Model group was decreased compared with PMP group?P<0.01?.CSS group was increased compared with model group?P<0.01?.9.In terms of Occludin mRNA expression of rats'colon tissues,model group was decreased compared with PMP group?P<0.01?,CSS group was increased compared with model group?P<0.05?.In terms of ZO-1 mRNA expression of rats'colon tissues,model group was decreased compared with PMP group?P<0.05?,CSS group was increased compared with model group?P<0.01?.In terms of expression of Occludin and ZO-1 protein of rats'colon tissue,compared with sham group,PMP group had a decreased trend in Occludin,and the ZO-1 decreased significantly?P<0.01?.Compared with PMP group,model group decreased significantly in both Occludin and ZO-1?P<0.01?.CSS group increased significantly compared with model group in both?P<0.01?.10.In terms of serum IL-1?of rats,PMP group was showed an increasing trend compared with sham group,model group was increased compared with PMP group?P<0.05?,CSS group was showed a downward trend compared with model group.In terms of serum IL-6 of rats,PMP group was increased compared with sham group?P<0.05?,model group was increased compared with PMP group?P<0.01?,CSS group was decreased compared with model group?P<0.05?.In terms of serum TNF-?of rats,PMP group was increased compared with sham group?P<0.01?,model group was increased compared with PMP group?P<0.01?,CSS group was showed a downward trend compared with model group.Conclusions1.Feed consumption reduction and weight loss were showed in rats of model group.2.The mass loss caused by liver-stagnation in perimenopausal rats were closely related to the gastrointestinal dysfunction caused by decrease of gastrointestinal hormones,increase of inflammatory factors,increase of intestinal mucosal permeability,decrease of gastrointestinal motility,and imbalance of intestinal flora.3.CSS could improve the conditon of liver-stagnation in liver-stagnation Pattern of perimenopausal rats,through up-regulate the gastrointestinal hormones,down-regulate the inflammatory factors,inhibit the imbalance of intestinal flora,and thus restore the gastrointestinal function.