| ObjectivesSlow transit constipation(STC)is a common clinical gastrointestinal disease attributed to ineffective colonic propulsion and/or increased resistance to propagation of colonic contents.The gradually rising prevalence rate of STC,which increases with age,is due in large part to the changes of people's life style.Constipation not only seriously affects the quality of life,and also has a close positive relationship with acute cardiovascular and cerebrovascular disease,dementia,colorectal cancer and so on.Aloe is rich in anthraquinone compounds,polysaccharides,amino acids,trace elements,and other variety of chemical constituents.Although aloe for the treatment of constipation have a long history,the mechanism of promotion of intestinal movement mechanism still remains unclear,particularly regulating gastrointestinal hormones have not yet reported.Based on the above points,aloin-the active ingredients of aloe was determined by HPLC,then slow transit constipation animal model was induced by Compound Diphenoxylate and the mechanism of preventive and the therapeutic effect of aloe on STC was observed.We also focus on the different changes of gastrointestinal hormone between STC animal and those treated with aloe in this study.These results might be beneficial to the measures of prevent and treat slow transit constipation.Methods1 Determination of aloin level in different aloe productThe aloin level in different aloe product were determined by HPLC with the following determination conditions:ZORBAX SB-18 column(4.6mm×150mm,5μm),the detection wavelength was 355 nm,the mobile phase of acetonitrile:water (25:75),flow rate:1ml/min,use external reference method to determine aloin in different aloe products by HPLC. 2 Screening on facitating feces excretion functions of different aloe productsThe aloe products(1.0 g/kg·bw)were given mice in advance,then Compound Diphenoxylate(10 mg/kg·bw)was used to induce constipation animal model.The first dejection time,stool amouts within six hours,feces traits,wet and dry faeces weight were observed.3 Effects of aloe on defecation and intestinal motor function of mice3.1 Effects of aloe on defecation function of normal mice:50 male Kunming mice were randomly divided into five groups,i.e.control group,aloe low,middle and high dose group(0.17 g/kg·bw,0.33 g/kg.bw,1.0 g/ kg·bw)and the positive control group(cisapride,2.5 mg/kg·bw).The first dejection time,stool amours within six hours,feces traits,fresh and dry faeces weight were observed after administration,and then fecal water content was calculated.3.2 Effects of aloe on defecation function of costive mice:60 male Kuning mice were randomly divided into 6 groups,aloe low,middle and high dose group(aloe 0.17 g/ kg·bw,0.33 g/kg·bw,1.0 g/kg·bw),the positive control group(cisapride,2.5 mg/ kg·bw),the blank control group and the model control group.Mice of Aloe low, middle and high dose group and the positive control group were given the corresponding dose aloe by gavage for seven days,the mice in blank control group and the model control group given the same volume of distilled water.On the 8thday the mice in model group,positive control group and aloe low,middle and high dose groups were treated with Compound Diphenoxylate(5 mg/kg·bw),mice of the blank control group with the same volume of distilled water.After treating with Compound Diphenoxylate 0.5 hours,aloe low,middle and high dose group and the positive control group were given the ink mixed with corresponding tested samples,while other mice only ink.The first dejection time,stool amours within six hours,feces traits,fresh and dry faeces weight were observed and recorded after Diphenoxylate administration,then fecal water content were calculate.3.3 Effects of aloe on intestinal motor function of mice:Animal groups and the administration method were same as 3.2 After giving ink 0.5 hour,blood were collected from inner canthus and content of NO were determined by using method of nitrate reductase,then mice were sacrificed,the entire intestine were quickly removed, and the intestinal transit ratio were calculated. 4 Effects of aloe on prevention and treatment of slow transit constipation of rats4.1 Effects of aloe on prevention of slow transit constipation of rats:48 male Wistar rats were randomly divided into four groups:control group,model group,low-dose aloe group and high-dose aloe group,with each 12.Model group,low-dose aloe group and high-dose group rats were given Compound Diphenoxylate 8 mg/ kg·bw by gavage,blank control group given the same volume of distilled water.Two hours later, the low-dose aloe group rats were given aloe 0.33 g / kg·bw,and high-dose group 0.66 g / kg·bw,while model group and the blank control group given the same volume of distilled water.Administration lasted eight weeks,gastrointestinal transit time and faeces weight were observed and recorded every fortnight.Eight weeks later, 8 rats in each group were killed and plasma and tissue specimens were collected in order to determine motilin(MTL),substance P(SP),vasoactive intestinal peptide (VIP)in plasma and colon.The rest rats were perfusion-fixed with paraformaldehyde for pathological examination.4.2 Effects of aloe on treatment of slow transit constipation of rats:48 Wistar rats were randomly divided into 2 groups:blank control group(n=12),STC model group (n=36).The rats of STC model group were given Compound Diphenoxylate 8 mg/ kg·bw by gavage every day for 8 weeks,blank control group given the same volume of distilled water.Eight weeks later,gastrointestinal transit time and faeces weight were observed and recorded.STC model rats were randomly divided into 3 groups: natural recovery group,low-dose aloe group,high-dose aloe group,with 12 each group.The 10thweek,the low-dose aloe group rats were given aloe 0.33 g/kg·bw, and high-dose group 0.66 g / kg·bw,while model group and the blank control group mice given the same volume of distilled water.Two weeks later,8 rats in each group were killed and plasma and tissue specimens were collected in order to determine motilin,substance P,vasoactive intestinal peptide in plasma and colon.The rest rats were perfusion-fixed with paraformaldehyde for pathological examination.Results1 Determination of aloin level in different aloe product:the aloin level in Aloe vera whole leaf powder is 0.088%,refined powder 0.266%,lyophilized powder leaves 0.908%,the whole foliar spray powder 6.71%,purified aloe powder for medicine 13.97%,aloe capsule 3.41%. 2 Screening on facitating feces excretion functions of different aloe products:Aloe refined powder and purified aloe powder for medicine could shorten the first dejection time,increase stool amouts and weight within six hours.Aloe leaves freeze-dried powder could shorten the first dejection time,increase stool weight.Aloe leaves sprayed powder could increase the weight of the stool.The whole aloe leaf powder has no obvious effects on the constipation.Among all the aloe products that were tested,aloe refined powder was found to be the most effective treatment on constipation.3 Effects of aloe on defecation and intestinal motor function of mice3.1 Effects of aloe on defecation function of normal mice:Aloe could shorten the first dejection time,increase stool amouts and weight,also could increase fecal water content.There were no significant effects on intestinal motor function to be observed in mice treated with Cisapride.3.2 Effects of aloe on defecation function of mice with constipation:aloe could improve stool traits.Compared with constipation model group,aloe in from 0.17g / kg·bw to 1.0 g / kg·bw dose could shorten the first black stool time and increase stool grains and weight in 6 hours of mice with constipation.3.3 Effects of aloe on intestinal motor function of mice:Compared with constipation model group,the propulsion rate of intestines in the aloe group was significantly higher than that of model group as well.The NO level in high-dose aloe group decreased obviously compared with model group(P＜0.05),and there was a negative correlation between the NO level of serum and propulsion rate of intestines in mice (r=-0.346,P＜0.05).4 Effects of aloe on prevention and treatment of slow transit constipation of rats4.1 Effects of aloe on prevention of slow transit constipation of ratsCompared with normal control group,STC model group rats had a longer first black stool time,less stool grains and decreased fecal water content,which showed STC model is induced successful.Compared with the STC model group,aloe prevention group could shorten the first black stool time and increase stool grains and fecal water content.Compared with normal control group,MTL decreased in intestinal tissue in the rats of STC model group(P＜0.05).Compared with the STC model group,high dose aloe could increase MTL content(P＜0.05).Compared with normal control group,SP content decreased in plasma and intestinal tissue in the rats of STC model group. Compared with the STC model group,low dose aloe could increase plasma SP content(P＜0.05),high-dose aloe could increase plasma and intestinal tissue SP content(P＜0.01).Compared with normal control group,VIP decreased in plasma and intestinal tissue in the rats of STC model group(P＜0.01).Compared with the STC model group,the low-dose aloe could increase intestinal tissue VIP content(P＜0.05), high-dose aloe could increase the plasma and intestinal tissues VIP content(P＜0.05).4.2 Effects of aloe on treatment slow transit constipation of rats.The results showed that compared with normal control group,STC model extent first black stool time(P＜0.01),decrease defecation grains(P＜0.01).Compared with the STC model group,aloe group could shorten the first black stool time and increase stool grains and fecal water content(P＜0.01).Compared with STC model group,MTL content increased in the high-dose aloe group(P＜0.05).Compared with normal control group,SP decreased in plasma and intestinal tissue in the rats of STC model group(P＜0.05,P＜0.01).Compared with STC model group,high-dose aloe could increase the plasma and intestinal tissues SP content(P＜0.01).Compared with normal control group,VIP decreased in plasma and intestinal tissue in the rats of STC model group(P＜0.01).Compared with STC model group,VIP levels in the intestinal tissue and plasma both in low-dose and high-dose aloe group increase,and high-dose aloe group has a statistically significant(P＜0.05).Conclusions1 Aloe products had the facitating feces excretion function,and which might be related to the processing methods and other factors.Refined aloe powder has a better role.2 To the normal mice,given the aloe of 1.0g/kg·bw to had the facitating feces excretion function,while to the constipation mice,0.17g / kg·bw had the same effect.3 Aloe could promote the mobility of intestine and ameliorate the constipation induced by Compound Diphenoxylate of mice,which might attribute to the increase of fecal water content and decrease of the serum NO level.4 Aloe had effect in the prevention and treatment of STC developed by Compound Diphenoxylate,which may be related to raise the MTL,SP,VIP content in plasma and intestinal tissue. |
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