Studies On The Safety And The Preventive And Therapeutic Effects Of Traditional Prescription Alatanqiqige-3 On Rat Model Of FD Based On Mongolian Medicine Theory

Objective The safety of Alatanqiqige-3 was evaluated by acute toxicity test in mice.The FD animal model preliminarily,which was in accordance with the etiology,pathogenesis and symptoms of'xila'Diffusion Syndrome by'Heyi'was established,and Alatanqiqige-3was used for treatment to evaluating model and to exploring the efficacy of Alatanqiqige-3.Methods 1.Determinated the oil content after processing the Momordica Cochinchinensis.According to 1:2.5:5 mix processed Momordica Cochinchinensis which was Within the range of oil content of processed products and Terminalia chebula,and the buds of Rosa rugosa Thunb.to the powder without agglomerate.2.80 healthy SPF mice,half male and half female,were randomly divided into 7administration group and?group A-G?and 1 control group?Group H?,10 in each group.The maximum lethal dose(D_max)and the minimum lethal dose(D_min)of single dose were obtained through the pre experiment.Then,the single dose of each group was calculated according to the K value.The dose was given twice in 0.4ml/10g volume with an interval of6h.The control group was given 1%CMC-Na solution twice in equal volume with an interval of 6h.The changes of body weight,poisoning and pathological changes of important organs were observed,Then calculate LD₅₀.3.SD male rats were divided into control group?10 rats?,model group?14 rats?,domperidone treatment group?14 rats?,Alatanqiqige-3 high dose group?14 rats?,Alatanqiqige-3 medium dose group?14 rats?,Alatanqiqige-3 low dose group?14 rats?.The control group had no stimulation.The other groups were given the following 7 stimulate:Fasting for 24 hours,water prohibition for 24 hours,tail clamping for5 minutes,45?environment for 30 minutes,Running on the treadmill for 30minutes,light and dark inversion?12/12 h?,scaring for 45 minutes,giving a stimulus at random every day,make the occurrence of the stimulus can not be expected,and the stimulus mode is different for two consecutive days.7 species were stimulated for 21 days.Except for the control group,rats in the other groups were given 200ml strong dark tea every other day for10 days.At the same time,the high dose group,middle dose group and low dose group of Alatanqiqige-3 were given 9g/70kg×6.3×2=1.62g/kg,9g/70kg×6.3=0.81g/kg,9g/70kg×6.3/2=0.405g/kg respectively.The domperidone treatment group was given domperidone 0.43mg/70kg×6.3=0.0387mg/kg suspension.The control group and model group were given 1%CMC-Na solution of equal volume every day.During the experiment,the appearance,tongue,feces and mental state of rats were observed,and the body weight,body temperature,record the amount of food and drinking water.After 21 days,the nutrition semi-solid paste was given on an empty stomach.10 minutes later,the stomach and intestine were anesthetized and the residual rate and intestine propulsion rate were measured.Blood was collected from abdominal aorta,then centrifuged,take the supernatant,and after gastric and intestinal residues were removed,carry out tissue homogenate.The levels of GAS,MTL,PP,CCK,ACh,5-HT,NPY,NO and VIP in serum and gastrointestinal tissues were measured.The liver,stomach and intestine of rats were taken for routine pathological sections,and the pathological changes were observed.Results 1.The oil content of the processed products was 27.35%,which was in the range of25%?30%,then the uniform powder of Alatanqiqige-3 was obtained.2.In the acute toxicity experiment,within 4 hours after administration,some mice showed decreased spontaneous activity,shrinking but no mice died.Mice died 6 hours after administration,and no mice died 76 hours later.After 3 days of observation,the feeding and drinking water of the surviving mice basically returned,weight began to grow steadily to normal,and there was no other abnormal phenomenon on 7 days of observation.The morphology,color and texture of heart,liver,spleen,lung and kidney of all mice were normal,and no obvious pathological changes were observed by naked eyes.Histopathological observation showed that:The degree of vacuolation of hepatocytes in the administration group was higher than that in the control group,In group B,there was a little pigmentation in the liver,and in group G,there was a little oval cell proliferation in the portal area.A small number of cells in the spleen were punctate necrotic and the nuclei were broken.There was no obvious pathological changeswere found in lung,kidney and heart.The LD₅₀ was 16.03 g·kg-1 and 95%CI was 11.38-22.60 g·kg-1.3.Pharmacological test results:?1?appearance:In the model group,the hair was disordered,yellow and dul,dry tongue with yellow and white tongue coating.Compared with the model group,the hair of the domperidone treatment group and the middle dose group was smooth without caking,pink,moist tongue with whitish tongue coating.There was no significant difference in the hair of high and low dose group compared with the model group,the tongue was dark pink,moist,with little white coating.?2?Mental state:The rats in the control group had good mental state,sensitive response,arounded the ice bags under high temperature,tends to be cool.In the model group,the rats became sensitive and easily frightened,sat away from the ice bag under high temperature,which was not like to cool.The mental state of the administration group was significantly better than that of the model group,had good mental state,sensitive response and arounded the ice bags,while the number of rats in high dose group was relatively less than that in the other Administration group.?3?Faeces:The control group had a large amount of faeces,normal size,soft texture,covered with water and sticky;The faeces became smaller,hard and dry in model group,and the amount of faeces decreased significantly?P<0.05?;The faeces was normal,soft and sticky in domperidone treatment group,and the amount of faeces increased significantly?P<0.05?;The faeces in three dosage groups was larger,some of faeces was soft and sticky,the amount of faeces increased,but have no significant difference?P>0.05?.There was no significant change in the color and quantity of urine in the six groups.?4?Body weight and temperature:The body weight of the model group increased slowly and significantly?P<0.05?,the body temperature showed irregular and unstable changes and the amplitude was large;The body weight of the domperidone treatment group increased significantly?P<0.05?;The change of body temperature is small and relatively stable,but there is no significant difference?P>0.05?;The body weight was no significant difference in three dose groups of Alatanqiqige-3?P>0.05?,the change of body temperature is small and relatively stable,but there is no significant difference?P>0.05?.?5?Diet and water intake:The amount of diet intake began to decrease in the second week,but appeared to return to normal in third week in model group and three dose groups of Alatanqiqige-3;The diet of domperidone group was without obvious change;There was no significant difference in the amount of drinking water between the four administration groups in the first two weeks and the model group?P>0.05?,and there was an increasing trend after two weeks.?6?Gastric residual rate and intestinal propulsive rate:Compared with the control group,the gastric residual rate of the model group was significantly increased,and the intestinal propulsive rate was significantly slower?P<0.05?;Compared with the model group,the gastric residual rate of the domperidone treatment group was significantly reduced,and the intestinal propulsive rate was significantly increased?P<0.05?;Compared with the model group,the gastric residual rate of three doses of Alatanqiqige-3 decreased and the intestinal propulsion rate increased,but no significant difference?P>0.05?.?7?Neurotransmitter ACh and NPY:Compared with the control group,the serum and gastrointestinal tissue ACh and NPY levels in the model group were significantly reduced?P<0.05?;Compared with the model group,the serum and gastrointestinal tissue ACh and NPY levels in domperidone treatment group and three doses of Alatanqiqige-3 were increased,but no significant difference in serum ACh level in the low dose group?P>0.05?.?8?Neurotransmitter 5-HT,NO,VIP:Compared with the control group,the serum and gastrointestinal tissue 5-HT,NO,VIP levels in the model group was increased,but no significant difference in serum NO level?P>0.05?;The serum and gastrointestinal tissue 5-HT,NO,VIP levels in the domperidone treatment group was decreased,but no significant difference in gastrointestinal tissue 5-HT,VIP levels?P>0.05?;The levels of 5-HT,NO and VIP in serum and gastrointestinal tissues were decreased in three doses of Alatanqiqige-3,but no significant difference in VIP level in the low dose group?P>0.05?.?9?Digestion related indexes:Compared with the control group,the levels of serum and gastrointestinal tissue PP,GAS,MTL and CCK in the model group were significantly decreased?P<0.05?,and the levels of serum and gastrointestinal tissue PP,GAS,MTL and CCK in each administration group were increased,but no significant difference in PP level in the low dose group?P>0.05?.?10?Histological changes:The structure of liver lobule,stomach and small intestine were clear,arranged orderly,and no obvious pathological changes were found.Conclusion 1.When the single use of Alatanqiqige-3 is too much,it will have a certain degree of influence on the liver and spleen,but it's safe to use in the specified dose range.2.The experimental results showed that the symptoms of the rats were in accordance with the symptoms of'xila'Diffusion Syndrome by'Heyi'and FD,as well as the standard of FD animal model.Therefore,the FD animal model based on Mongolian medicine theory has been established successfully.3.Through the pharmacodynamic experiment,Alatanqiqige-3can effectively improve the symptoms and signs of rats;At the same time,Alatanqiqige-3can increase the levels of digestive enzyme PP,digestive related hormones GAS,MTL,CCK and neurotransmitter ACh,NYP,and reduce the levels of neurotransmitter 5-HT,VIP and NO in serum and gastrointestinal tissues.But has no effect on the liver,stomach and small intestine tissues of rats.Therefore,Mongolian medicine Alatanqiqige-3 has a significant effect on the prevention and treatment of FD model rats based on Mongolian medicine theory,among which the effect of middle dose group of Alatanqiqige-3 is the best,the mechanism may be related to regulating the neurotransmitters and digestive hormones,improving gastrointestinal peristalsis.