Research On The Total Phenolic Acid Extraction Of Chebulae Fructus And Its Antidiarrheal Properties

Chebulia is the dried ripe fruit of the Terminalia chebula Retz.or Terminalia chebula Retz.Var.tomentella Kurt.Taste Bitter,acid,acerbity and flat.The main effect of it is anti-cough and anti-diarrheal.Can be used for the treatment of continuous diarrhea of horses,cattle,sheep,pigs and other heavy livestocks.So far,there is no related reports about Chebulia antidiarrheal experiment,therefore,this study on the basis of the optimization of the ultrasonic extraction process of Chebulia total phenolic acids,evaluated the the anti-diarrheal activity of Chebulia,and further carried on the preliminary discussion of the anti-diarrheal mechanism of Chebulia based on ion channels.By using the combined method of single factor and response faces,we determined the ultrasonic extraction technology of Chebulia,the optimum conditions is as follows:68%ethanol concentration,ultrasonic intensity of 3.6 W/cm~2,solid-liquid ratio at 23 mg/mL,particle size of0.18 mm,20 min ultrasonic time,extraction temperature at 70℃,and extract 2 times.In the best condition,the extraction yield of total phenolic acids was 448.7±2.15 mg GAE/g DW close to the predicted value(447.8 mg GAE DW/g).Chebula Fruits extract inhibit production of DPPH and ABTS and super oxygen free radicals,which shows that the fruits possess strong reducibility and antioxidant capacity.The test also shown that,in a certain range of concentration,in vitro antioxidant activity is related with the content of total phenolic acids.The second part of this article,we studied the effect of Chebula Fruits aqueous extract and its different extraction section on antidiarrheal activity.Diarrhea model caused by castor oil in mice or rats was used to verify the Chebula Fruits aqueous extract of its antidiarrheal activity and to find out the active part of the extraction.Verify the effect of Chebula Fruits on small intestine and liver by histopathology.The dose for the trial was set at 200,400 and 800 mg/kg and it present a dose dependent antidiarrheal effect,of which the 400 and 800 mg/kg dose group significantly extend the onset of diarrhea,reduced the frequency of wet/loose stools and the propulsion of charcoal.Chebula Fruits aqueous extract(200,400 and 800 mg/mL,oral)significantly(p<0.05)and dose-dependently reduced the castor oil inducted rat small intestine fluid secretion.Small intestine and liver of castor oil treated mice appeared significantly pathological changes.However,the damage of mice treated with medicine diminished.In addition,the ethyl acetate fraction of Chebula Fruits aqueous extract has significant(p<0.05)antidiarrheal activity,it delayed the onset of diarrhea and reduced the rate of loose stools.The main composition of ethyl acetate fraction is tannins,and the major components of it was shikimicacid,gallicacid,5-O-galloylshikimicacid,corilagin,3,4,8,9,10-Pentahydroxy-dibenzo[b,d]pyran-6-one and ellagic acid by HPLC-ESI-MS analysis.Finally,the article has carried on a preliminary inquiry of Chebula Fruits antidiarrheal mechanism,and the diarrheal was caused by castor oil.Generally,BALB/c mice were divided into 4 groups,administered orally medicine for seven days,then the mouse was sacrificed,the middle part of the jejunum was removed for investigation.application fluorescence quantitative PCRmeasure the variation of exoression quantity of related six ion channel:NHE1,NHE2,NHE3,CFTR,SGLT1 and AQP4.Experimental results showed that:compared with the blank group,the NHE1,NHE2,CFTR,SGLTI and AQP4 ion channel gene transcription level of diarrhea model group were suppressed obviously;Compared with the diarrhea model group,the CFTR,NHE1 and SGLT1 gene expression standard of the treatment group rised in a certain degree,which proved that the Chebula Fruits extracts play a role in the three ion channels.This experiment determined the extraction process of total phenolic acids from Chebulia,and evaluated the antidiarrheal activity of the medicine.Carried on the mechanism of Chebulia anti diarrheal activity.It provided necessary theoretical basisas for the application of antidiarrhoeal medicines in clinic.