Preparation Of Galla Chinensis Oral Solution As Well As Its Secretory Diarrhea Inhibition Activity Evaluation

Galla chinensis contains a generous amount of tannins,has been used for a long time for the treatment of diarrhea,prolonged coughing and spontaneous perspiration in traditional Chinese medicine.In fact,many medical plants which are rich in tannins were proved effective in diarrhea.However,there are few detailed studies about pharmaceutical preparations of Galla Chinensis tannin extract.In the present experiments,for better application and to find the possibility that Galla Chinensis tannin extract can be used as an antidiarrheal drug,we prepared Galla Chinensis oral solution?GOS?and evaluated its antibacterial and antidiarrheal activities.The methods and results are as followed:1.Preparation of Galla Chinensis oral solutionThe tannin extract was extracted from Galla Chinensis,and the tannin content of the extract was measured.GOS was prepared with the tannin extract,purified water,appropriate excipients?sucrose,benzoic acid,ethyl p-hydroxybenzoate and natrium pyrosulfurosum?,and evaluated its physicochemical and microbiological stability at two conservation conditions for six months and twelve months in unopened brown bottles.As results,The tannin content of the extract was 47.75%.In accelerated stability test,after storage for six months and twelve months in different temperature conditions,the tannin content of GOS was still more than 90%,60mg/mL.2.The antibacterial activity of GOS in vitro and in vivoThe in vitro studies were reflected by minimum inhibitory concentration?MIC?and minimum bactericidal concentration?MBC?against Escherichia coli which is the causative agent for diarrhea.The in vivo study,mice were infected with LD₅₀0 of E.coli O101.And the body weight,survival rate,organ cofficients,levels of cytokine and immunoglobulin,cecum bacteria flora,and histopathological examination in colon of mice were evaluated.The MIC and MBC values were ranging from 1.17 to 2.34 mg/mL and 9.38mg/mL,respectively.In vivo studies supported that the survival rates of GOS-test groups are more than 50%,indicating that GOS could inhibit the infection of E.coil O101;there was a very significant difference in small intestine coefficient between negative control and other groups?P<0.01?,GOS 10%and GOS 15%had no significant difference with loperamide?P>0.05?,indicating that the GOS could decrease the fluid secretion of E.coli O101 infection in mice;results suggested that GOS improved the inflammation status of the infected mice by reducing the levels of pro-inflammatory cytokines like interferon??IFN-??,TNF-?,interleukin 1??IL-1??,IL-6,IL-8,and by increasing the levels of the anti-inflammatory cytokine IL-4;The IgG and sIgA levels were significantly higher?P<0.05?than those in negative control on day 4 or day 7,suggesting that GOS could increase the concentrations of immunoglobulin,like IgG and sIgA,to inhibit the infection of E.coil O101;GOS could increase the probiotics number,when the balance of the intestinal flora has been destroyed3.The antidiarrheal activity of GOS in miceThe antidiarrheal activity was determined in a castor oil-induced diarrhea model in mice through diarrhea symptoms,fluid accumulation ratio and percentage of distance moved by charcoal meal.GOS intervention resulted in prevented the structure of mucous layer and showed a better effect on the structure of mucous layer recovery.Meanwhile,GOS showed significant antidiarrheal activity in a castor oil-induced diarrhea model in mice?P<0.01?.In conclusion,the tannin content in GOS was not affected by prolonged?6months and 12 months?storage.GOS has therapeutic and protective effect to inhibit E.coil O101 infection by altering the levels of cytokines and immunoglobulin,regulating the intestinal bacteria and protecting the structure of colonic mucosa.and its antidiarrheal activity against castor oil-induced diarrhea in mice through alleviating diarrhea symptoms and inhibiting small intestinal motility and secretion.The results suggested that GOS is an effective and stable antidiarrheal drug that can be used to complement other therapies.