Effects Of Yujin Powder On Gastrointestinal Hormone And Hypothalamus-pituitary-adrenal Axis Of Large Intestine Dampness-heat Syndrome Rat Model

Large Intestine Dampness-heat Syndrome?LIDHS?of the livestock and poultry mainly has the symptoms of abdominal pain,diarrhea,bloody mucopurulent stools,fever and thirsty,rapid and slippery pulses,yellowish tongue.It is a common syndrome and is seen acute gastroenteritis and bacillary dysentery.And dampness-heat type diarrhea is one of the major types,the incidence of which is rising with the increase of intensive farming and mobility of livestock and poultry such as pigs,cattle and chickens.It has caused huge economic losses to the breeding industry.According to clinical practice showing,Yujin Powder?YJP?,one classical prescription in Traditional Chinese Veterinary Medicine?TCVM?,has good effects on LIDHS.This study is based on the determination of active components of YJP,the rat model of LIDHS was established by complex factors.It was treated with high,middle and low dose of YJP.In order to explore the essence of the LIDHS and the treatment mechanisms of YJP from gastrointestinal hormones and HPA axis,the contents of gastrointestinal hormone in serum and ileum and colon tissue,adrenal index and the contents of CRH,ACTH and CORT were detected.Results were as follow:1.Determination of active contents in YJP A high performance liquid chromatography?HPLC?method was established for simultaneously determining seven active components---baicalin,geniposide,paeoniflorin,gallic acid,berberine,curcumin and emodin in YJP.The seven active components of YJP were separated on ZORBAX SB-C18 column?4.6×150 mm,5?m?at a column temperature of 25?.The mobile phases were composed of?A?aqueous phosphoric acid?0.2%,v/v?and?B?methanol,?A?aqueous acetic acid?10%,v/v?and?C?acetonitrile with gradient elution at the flow rate of 1.0 mL/min.The three detection wavelengths were set at 240,275and 425 nm,respectively.The injection volume was 20?L.The results showed that the mass concentrations of baicalin,geniposide,paeoniflorin,gallic acid,berberine,curcumin,and emodin were as follow respectively:1.373²1.24?g.mL^-1?r=0.9994?,1.438²3.00?g.mL^-1?r=0.9999?,1.900³0.50?g.m L^-1?r=0.9998?,10.00¹61.0?g.mL^-1?r=0.9996?,11.90¹90.0?g.m L^-1?r=0.9998?,0.00748⁰.036?g.mL^-1?r=0.9994?,0.0038⁰.019?g.mL^-1?r=0.9991?.All calibration curves showed good linear regressions?R²>0.9991?within test ranges.The results of recovery test were99.9%-103.6%with a RSD value of 0.3%-1.7%.This method was simple,reliable and controllable,which can be used to quickly detect the content of 7 active components in YJP.2.Establishment of LIDHS model and observation of general behavior Sixty Wistar rats?half males and half females?were randomly divided into normal control group,LIDHS model group,self-healing group,high,middle and low dose of YJP group.The LIDHS model was established through the following steps:let the rats drink honey water freely;in the first 10 days,fasting and sufficient feed combined with gavage of lard were alternately performed;the rats were gavaged with wine and put in high temperature and high humidity environment(8h·d^-1)during the following 5 days,then they were injected with Escherichia coli once again after 24 hours,After the establishment of the model,the rats were treated with high,middle and low dose of YJP,respectively.During the experiment,the clinical symptoms and signs of rats were observed.Then blood routine,blood biochemistry,serum inflammatory cytokines?TNF-?,IL-1?,IL-6,IL-10,IL-17 and IL-23?levels and the histopathological changes of ileum and colon were detected and observed,respectively.The results showed that the clinical symptoms and signs of the model rats were consistent with the diagnostic criteria of LIDHS,moreover,there was obvious systemic inflammatory response,meanwhile the blood lipid increased,intestinal mucosa was damaged and extensive congestion.And after treatment with YJP in different dosages,the clinical symptoms and signs of the rats with LIDHS were improved;the indexes of blood routine,blood biochemistry and inflammatory cytokines levels tended to be normal,and histopathological changes of ileum and colon were alleviated.There was an obvious dose-effect relationship,in which the effects of high dose was the best.3.The studies of gastrointestinal hormones in serum and intestinal tissue For each group,the contents of gastrointestinal hormones in serum,ileum and colon tissue were detected.Results were as following:Compared with normal group,the contents of motilin?MTL?,gastrin?GAS?,substance P?SP?increased significantly?P<0.01 or P<0.05?in serum,ileum and colon tissue in model and self-healing group.Somatostatin?SS?decreased significantly?P<0.01?and vasoactive intestinal peptide?VIP?showed no significant changes.Compared with self-healing group,after treatment with various doses of YJP,the above-mentioned indexes tend to normal condition to varying degrees,especially in the high dose group.In the high dose group,the contents of MTL,GAS and SP decreased significantly?P<0.01 or P<0.05?,and SS increased significantly?P<0.01 or P<0.05?in serum,ileum and colon tissue.4.Research on HPA axis function For each group,adrenal index and the contents of CRH,ACTH and CORT in serum were detected.Results were as following:Compared with normal group,adrenal index and the contents of CRH,ACTH and CORT in serum increased significantly?P<0.01 or P<0.05?in model and self-healing group.Compared with self-healing group,after treatment with various doses of YJP,the above-mentioned indexes tend to normal condition to varying degrees,especially in the high dose group.In conclusion,in this study,the method was established for simultaneous determination of 7 active components in YJP by HPLC.The rat model of LIDHS was successfully established,YJP had a good effect on it by anti-diarrhea,anti-inflammatory,lowing blood lipid,protecting intestinal mucosa,adjusting the disorder state of gastrointestinal hormones and HPA axis function.