Effect Of Hawthorn Seeds Extract On Gastrointestinal Function And Preparation Of Effervescent Tablets

Objective: The hawthorn seed is the seed of hawthorn(Crataegus pinnatifida Bunge),a Rosaceae plant,which is rich in various chemical components.However,there are relatively few studies on the extraction process,activity and preparation of hawthorn seeds.In this study,the extraction technology of hawthorn seeds was optimized,its effect on promoting gastrointestinal motility was studied,the optimal active site was screened,its therapeutic effect and potential mechanism on diabetic gastroparesis rats were explored,and solid dispersion effervescent tablets of hawthorn seeds were prepared.Methods: Ethanol reflux extraction was used to optimize the extraction process by single factor screening combined with response surface methodology,using total phenolic acid yield as the index.Hawthorn seeds were extracted by the optimum extraction process,and then petroleum ether,ethyl acetate and n-butanol were used to extract the hawthorn seeds.The effects of different extracts on gastric emptying,small intestine propulsion in mice and gastric juice secretion,total acid excretion and pepsin activity in rats were detected to determine the optimum active site.A rat model of diabetic gastroparesis(DGP)was established to explore the effects of different doses of hawthorn seeds ethyl acetate extract(HEA)on body weight,blood glucose,gastric emptying and small intestine propulsion in DGP rats.The effects of HEA on superoxide dismutase(SOD),malondialdehyde(MDA)and glutathione peroxidase(GSH-Px)in gastric homogenate were measured.The levels of ghrelin,motilin(MTL)and gastrin(GAS)in rat plasma were determined by radioimmunoassay and ELISA,respectively.Western Blot method was used to detect the expression of c-kit,ghrelin and neuronal nitric oxide synthase(nNOS)in the stomach,and immunohistochemical method was used to detect the changes of interstitial cells of Cajal(ICC)in the stomach.Therefore,the mechanism of HEA on DGP rats was explored.With HEA as the main drug and the cumulative dissolution rate of solid dispersion as the index,the preparation methods,carrier types and drug loading ratio of solid dispersion were screened to determine the optimum preparation process of hawthorn seeds solid dispersion.The formulation of effervescent tablets was optimized by single factor test and orthogonal test,using disintegration time,pH value,hardness,foaming volume and sensory evaluation as evaluation indexes.The hawthorn seeds solid dispersion effervescent tablets were prepared by polyethylene glycol(PEG)encapsulation of alkali source and wet granulation and tablet pressing method,and the quality test and acute oral toxicity test were carried out.Results: The optimal extraction technology of hawthorn seeds was 70% ethanol concentration,2.5 h reflux time,22 mL/g liquid-solid ratio,86 ? extraction temperature,twice extraction.Under these conditions,the total phenolic acid content of hawthorn seeds was 4.83 mg/g,and the process was stable and reliable.Compared with the control group,the ethyl acetate fraction of hawthorn seeds had the most significant effect on gastric emptying and small intestine propulsion in normal mice(P<0.01),which was similar to the positive drug morpholine(P>0.05).Meanwhile,the ethyl acetate group had the most strongest effect on gastric juice,total acid excretion and pepsin activity(P<0.01).Different doses of HEA can reduce blood sugar in DGP rats to a certain extent,and significantly inhibit the weight loss of DGP rats.Furthermore,it effectively promoted the gastric emptying and small intestine propulsion(P<0.05 or P<0.01)with a dose-dependent manner.The effect of high doses of HEA is similar to that of positive drugs(P>0.05).In addition,HEA increased SOD and GSH-Px in stomach while decreased MDA,increased plasma ghrelin while decreased MTL and GAS in a dose-dependent manner(P<0.05 or P<0.01).The expression of c-kit,ghrelin and nNOS protein and the number of ICC cells in the stomach also significantly increased(P<0.05 or P<0.01).Hawthorn seeds solid dispersion prepared by solvent method,PVP K30 as carrier and 1:10 drug loading ratio significantly improved the solubility of HEA.The optimum formulation of effervescent tablets was determined as follows: 25% solid dispersion,47.5% total effervescent agent,1:1.5 tartaric acid and sodium bicarbonate and 1.5% aspartame.Conclusions: Hawthorn seeds promote the gastrointestinal motility and the main active part is ethyl acetate extract.HEA improves gastrointestinal motility in DGP rats,and its potential mechanism might be involved in the inhibition of oxidative stress injury induced by hyperglycemia,up-regulating n-NOS,c-kit expression and ICC celles in the stomach,regulating gastrointestinal hormones.The formulation of Hawthorn seeds solid dispersion effervescent tablets was reasonable with a feasible technological conditions.The product was qualified with excellent safety and reliability.