| Objective: Salsola collina is a genus Salsoloideae,widely distributed in saline and alkaline areas such as Central and Southwest Asia,the Mediterranean and North Africa.It is rich in a variety of chemical components,but there is no report on the activity and preparation of ethyl acetate extract of Salsola collina(EES).In this study,the composition of EES was studied,the effect and potential mechanism of treating diabetic gastroparesis(GDP)were discussed,and the preparation technology of its dispersible tablets was optimized.Methods: The alcohol extract of Salsola was prepared by ethanol reflux method,and EES was obtained by extraction.The composition of EES was analyzed by high performance liquid chromatography(HPLC)and high performance liquid chromatography-mass spectrometry(LC-MS).The DGP model was established by injecting streptozocin(STZ)into rats and feeding them with high sugar and high fat(hshf)feed irregularly.The changes of food intake,body weight and blood glucose were recorded.After the experiment,gastric emptying was measured by phenol red labeling.Determination of nitric oxide(no),total cholesterol(TC)and triglyceride(TG)in rat serum by enzyme-linked immunosorbent assay,as well as superoxide dismutase(SOD),malondialdehyde(MDA),catalase(CAT)and glutathione peroxidase(GSH-Px)content in gastric tissue.The expression of neuronal nitric oxide synthase(nNOS)and protein gene product 9.5(PGP9.5)were detected by Western blot and immunohistochemistry.The concentrations of ghrelin,gastrin(GAS),somatostatin(SS)and vasoactive intestinal peptide(VIP)in the serum of rats were determined by the kits.The pathological changes of gastric tissue were observed by hematoxylin eosin(HE)staining.The expression of c-kit and its natural ligand stem cell factor(SCF)in gastric tissue was detected by real-time polymerase chain reaction(Real-Time PCR),Western blot and immunofluorescence.The mechanism of EES on DGP rats was discussed.With EES as the main drug,the powder was pressed directly,and the appearance and disintegration time of the tablet were taken as the inspection indexes.Through single factor test and multi factor orthogonal test,the optimal preparation technology of the target dispersible tablet was determined,and its quality inspection,stability test and toxicity test were carried out.Results: EES is mainly composed of D-galacturonic acid,orsellinic acid,protocatechuic acid,caffeic acid,salicylic acid,vanillic acid,syringic acid,p-coumaric acid,ferulic acid and p-hydroxybenzoic acid.Compare with the DGP rats,food intake,weight and blood glucose in rats treated with different doses of EES were improved to varing degrees(P < 0.05-0.01);serum TC and TG decreased(P < 0.05-0.01)while serum NO increased(P < 0.05-0.01);The concentration of SOD,CAT and GSH-PX in gastric tissue increased,while the concentration of MDA decreased(P < 0.05-0.01);The expression of nNOS and PGP9.5 in gastric tissue increased significantly(P < 0.05-0.01).At the same time,serum ghrelin and GAS increased,while SS and VIP decreased after EES intervention(compared with DGP group,P < 0.05-0.01).EES can also restore the gastric injury and increase the expression of c-kit and SCF protein(compared with DGP group,P < 0.05-0.01).The best prescription of EES dispersible tablets was determined as follows: 30% for EES,37% for MCC and erythritol(10:8),30% for dry starch,PVPP and L-HPC(2:1:1),and 3% for micronized silica gel and magnesium stearate(5:2).The stability of EES dispersible tablets was proved to be good and safe.Conclusions: EES promotes gastric emptying and reduce gastric injury in DGP rats.Its mechanism may be related to its hypoglycemic and lipid-lowering activities,inhibition of oxidative stress and the increase of the number of gastric neurons.It is related to the regulation of gastrointestinal hormone secretion and c-kit / SCF signal pathway by EES.This multi-channel and multi-mechanism therapeutic effect makes it important to develop the application value of EES.The formulation of EES dispersible tablet is reasonable,the process conditions are feasible,the product quality is qualified,which is safe and reliable. |
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