| Objective:To study the effect of spleen-deficiency constipation model made on the intestinal microbiota,enzyme activities and blood routine in mice, provide basis on the treatment of spleen-deficiency constipation.The relationship between efficacy of ultra-micro powder Dendrobium officinale on the intestinal microbiota, enzyme activities, blood routine,molecular diversity of intestinal Bacteria and Lactobacillus in mice with spleen-deficiency constipation was studied to provide the basis on its mechanism of action.Methods:The mouse models of spleen-deficiency constipation were established with senna water decoction7days, then diet was restricted, to make them hunger satiety disordered. The mouse of nomal group were were given equivalent amount of sterile water, then were fed normally. The models were made in total15days. The body weight, intestinal microbiota, enzyme activities and blood routine were snalysed.The treatment groups were respectively given the traditional decoction of Dendrobium officinale,the full amount,50%amount,25%amount of ultra-micro powder Dendrobium officinale,Simo decoction,the normal group and the model group were given equivalent amount of sterile water.The intestinal microbiota enzyme and activities were snalysed after7days. Then treatment groups were respectively given the traditional decoction of Dendrobium officinale,50%amount of ultra-micro powder Dendrobium officinale,the normal group and the model group were given equivalent amount of sterile water. Adopted the eyeball blood collection method, and used the CA-500automated hematology analyzer for determination.The mouse with spleen-deficiency were constipation in treatment groups respectively given the traditional decoction of Dendrobium officinale,50%amount of ultra-micro powder Dendrobium officinale.The metagenome DNA of intestinal flora was extracted from intestinal contents. Then ARDRA analysis was made after PCR by universal primer of Bacteria and specific primer of Lactobacillus.Results:The results showed that amount and rate of the body weight change of mice in the model group were smaller than those in the normal group (P<0.05or P<0.01), compared to normal group the amount of bacteria was reduced (P<0.01), the amount of Escherichia coli, Lactobacillus, Bifidobacterrium, Fungus were increased. The activities of amylase, protease were receded (PO.05or PO.01), but xylase, cellulase were enhanced compared to normal group mice were different from the mouse in normal group (PO.01). The mice with spleen-deficiency constipation whose PLT (Platelet) and MPV (Mean Platelet Volume) were significantly lower than normal group (P<0.05). And WBC (White Blood Cell) was significantly higher than normal group (PO.05). The other indexes had no significant difference with normal group (P>0.05).Spleen-deficiency constipation had no significant improvement in model group, amount and rate of the body weight change of mouse were smaller than those in the normal group,the intestinal microbiota(ba.cteria,Escherichia coli, lactobacillus, bifidobacterrium, fungus) and the cellulase, xylase, amylase, protease were had significant difference with the normal group(PO.01or P<0.05).The spleen-deficiency constipation had significant improvement in all treatment groups,and whose lactobacillus,bifidobacterrium were more than the normal group(PO,01or PO.05). Escherichia coli reached the level of normal group in all treatment groups except the full amount of ultra-micro powder Dendrobium offidnale(P<Q.05). The fungus reached the level of normal group in50%amount of ultra-micro powder Dendrobium officnaler group and Simo decoction group.The activities of cellulase and amylase in full amount of ultra-micro powder Dendrobium officnaler group, cellulase in50%amount of ultra-micro powder Dendrobium officnaler group, protease in25%amount of ultra-micro powder Dendrobium officnaler group and Simo decoction group were reached the level of normal group(P>0.05). The activities of xylase in all treatment groups were lower than the normal group(P<0.01).After treatment with Dendrobium officnale, platelet(PLT) and thrombocytocrit(PCT) were significantly higher than normal group(P<0.01or P<0.05);red blood cell (RBC) and haematocrit(HCT)were increased; Rbc Volume Distribution Width (RDW), hemoglobin(HGB), white blood cell(WBC)and lymphocytotoxicity (LYM) were reduced;Mean Corpuscular Hemoglobin(MCH) and Mean Corpuscular Hemoglobin Concentration(MCHC) were also reduced,but still were significantly higher than normal group(P<0.01or P<0.05).The RBC and HCT of50%amount of ultra-micro powder Dendrobium officinale group were significantly higher than model group(P<0.05),reached the lever of normal group(i^>0.05).The WBC and LYM of50%amount of ultra-micro powder Dendrobium officinale group were significantly lower than model group and the traditional decoction of Dendrobium officinale group(P<0.01orP<0.05),but had no significant difference with normal group(P>0.05).The results showed that OTUs,Shanon index and Brillouin index in the nomal group, the50%volume of ultra-micro powder group were the same and larger than in the model group and the traditional decoction group. The similarity coefficient which compared with Bacteria community structure in normal group was the largest in the50%volume of ultra-micro powder group, then in the model group and the traditional decoction group. The cluster analysis showed that groups were clustered in2classes when the clustering distance>15and<25,the nomal group and the50%volume of ultra-micro powder group were in one class,then the others. All groups were clustered in4classes when the clustering distance<15,one group was in one class.The principal components analysis suggested that the nomal group and the50%volume of ultra-micro powder group were had greater contribution to PC1,and the model group and the traditional decoction group were had same contribution to PC2.The molecular diversity of intestinal Bacteria in mice changed by the affect of spleen-deficiency constipation model,and the two kinds of decoction of Dendrobium officinale regulated it in different ways. Study the effect of ultra-micro powder Dendrobium officinale on the molecular diversity of intestinal Lactobacillus in mice with spleen-deficiency constipation. The results showed that OTUs,Shanon index and Brillouin index in the nomal group,the traditional decoction group and the1/2volume of ultra-micro powder group were the same and larger than in the model group.The Czekanowski coefficient0.3333was the largestest in the1/2volume of ultra-micro powder group,0.1818in the model group,0.1667was the least in the traditional decoction group,compare with Lactobacillus community structure in the normal group.The cluster analysis and principal components analysis suggested that molecular diversity of intestinal Lactobacillus in mice changed by the spleen-deficiency constipation model made,and the two kinds decoction of Dendrobium officinale regulated it in different ways.Conclusion The spleen-deficiency constipation was made had affect the body weight, had destroyed the balance intestinal microbiota and influnce intestinal enzyme activities. The modeling spleen-deficiency constipation had significantly influence on PLT, MPV and WBC, which affected the function of blood system in mice. The traditional decoction of Dendrobium officinale, and the full amount,50%amount of ultra-micro powder Dendrobium officinale and Simo decoction had curative effect on spleen-deficiency constipation,regulated the balance of intestinal microflora and the intestinal enzyme activity through different pathways.The50%amount of ultra-micro powder and traditional decoction of Dendrobium officinale had the equal curative effect with Simo decoction. Dendrobium officinale had effect on the RBC,HCT,RBC,RDW,HGB,MCH, MCHC,WBCand LYM in mice with spleen-deficiency constipation,and the overall curative effect of50%amount of ultra-micro powder Dendrobium officinale was better than the traditional decoction.The molecular diversity of intestinal Bacteria and Lactobacillus in mice changed by the affect of spleen-deficiency constipation model,and the two kinds of decoction of Dendrobium officinale regulated it in different ways. The efficacy of50%volume of ultra-micro powder Dendrobium officinale was the better than the traditional decoction group. |
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