Research Of Effects And Mechanisms Of Different Extracts Of 5 Kinds Of Traditional Chinese Medicines Of Alpinia On Rats Induced Gastric Ulcer With Cold Syndrome

Objective: rats are administered by gavage with acetic acid and ice anemarrhena,while cold factors gave for the rats to copy gastric ulcer mo-del with cold syndrome,respectively taking four different polar fraction-s of five flavour herbals of Alpinia（petroleum ether,ethyl acetate,nbutanol and water extracts）to study the efficacy and mechanism of action.Observeing effects of different polar parts of five flavour Alpinia of Chinese traditional medicine for stomach tissue pathological changes,pepsin activity,testing stomach tissue c AMP and c GMP value,and calculate the c AMP/c GMP ratio,illustrating the actions of effective parts of five flavour herbals of Alpinia for rats made gastric ulcer with cold syndrome.At the same time,detecting values of EGF,PGE₂,TGF-alpha in gastric tissue,and detecting values of PAF,NO,ET,IL-8,TNF alpha,NT,VEGF,b FGF in serum,primarily study the mechanism of action of effective parts of five flavour herbals of Alpinia for gastric ulcer with cold syndrome.And interpretating the link between the law of "relationship、Chinese medicine efficacy and pharmacological effect",in order to further ascertain common effect of the material basis of five flavour herbals of Alpinia to provide the reference.Methods: 450 SD male rats,10 rats randomly were taken to blank group,others rats were filled with 15% of ice acetic acid and 4 ℃ rhizoma anemarrhenae decoction liquid to the stomaches for duplicating gastric ulcer model with cold syndrome,after the models copied successfully,setting the disease syndrome model group,2 positive control groups,40 subjects drug groups,each group of 10 rats.Blank group and model group were given 0.5% sodium carboxymethyl cellulose solution,the other groups were given the subjects medicine groups respectively.observing stomach tissue pathological changes after HE dyeing,determining pepsin activity;taking ELISA determination to detect values of PAF,NO,ET,IL-8,TNF alpha,NT,VEGF,b FGF in serum of rats,and PGE₂,EGF,TGF-alpha,c AMP and c GMP in stomach tissue,calculating the c AMP/c GMP ratio.SPSS20.0 is adopted to improve the statistics,with P < 0.05 that was statistically significant.Results: 1.The pharmacodynamics study of five flavour herbals of Alpinia on gastric ulcer（1）the effects of pathological changes of gastric mucosa in rats with gastric ulcer Compared with the model group,the rats gastric tissue pathological score of five flavour herbals of Alpinia in petroleum ether and ethyl acetate fractions are significantly more decreased（P< 0.01 or P < 0.05）;three flavour herbals of Alpinia in n-butanol site groups（Alpinia galanga,galangal fruit,Alpinia Katsumadai Hayata）,rats gastric tissue pathological score in water site grops（Alpinia galanga Willd,Alpinia Katsumadai Hayata,Alpinia oxyphylla）were lower（P<0.01 or P<0.05）,pathology of gastric tissue of rats in other groups were not significantly decreased.（2）The influence of pepsin activity Compared with model group,the high and low dose group of 5 flavor drugs were tested in the petroleum ether and ethyl acetate parts,the high dose group of 3 flavor drugs were tested in n-butanol part（Alpinia officinarum Hance,Alpinia galanga Swartz,fructus alpiniae galangae）,the high dose group of 3 flavor drugs were tested in water site（Alpinia officinarum Hance,Alpinia galanga Swartz,Alpinia oxyphylla）,the low dose group of Alpinia officinarum Hance in water site all could significantly lower values of pepsin activity（P< 0.01 or P<0.05）,the rest of the polarity parts each dose group rats pepsin activity change is not obvious.（3）The influences of values of c AMP、c GMP to rats made gastric ulcer with cold syndrome Compared with model group,petroleum ether and ethyl acetate parts of high dose group of five taste reagents,n-butanol part high dose group of three taste reagents（fructus alpiniae galangae,Alpinia katsumadai,Alpinia oxyphylla）in rats stomach tissue c AMP value（P<0.01 or P<0.05）;the difference of c GMP value in all of rats stomach tissue is not obvious,have no statistical significance（P>0.05）;petroleum ether part high dose group and the ethyl acetate parts of high dose group of five taste reagents,galangal fruit and fructus alpiniae oxyphyllae n-butanol part c AMP/c GMP ratio high dose group were significantly increased（P <0.05）.2.Studying of the mechanism about 5 kinds of Alpinia herbs on rats with gastric mlcer and cold Syndrome（1）Effects of different polar parts of chinese herbal medicine on the expression of TGF-α,PGE₂ and EGF in stomach tissue of rats with gastric ulcer and cold syndrome In terms of TGF-α,compared with the model group,5 flavor drugs were tested in four different polar parts of the high dose group,4 flavor drugs were tested in the low dose group of petroleum ether（Alpinia officinarum Hance,Alpinia galanga Swartz,fructus galangae,Alpinia oxyphylla）,3 flavor drugs were tested in the low dose group of ethyl acetate parts（Alpinia officinarum Hance,fructus galangae,Alpinia katsumadai）,3 flavor drugs were tested in the low dose group of n-butanol parts（Alpinia officinarum Hance,Alpinia galanga Swartz,Alpinia oxyphylla）,3 flavor drugs were tested in the low dose group of water parts（Alpinia galanga Swartz,Alpinia katsumadai,Alpinia oxyphylla）all coμLd enhance the expression of TGF-α in gastric tissue,the difference was significantly obvious（P<0.01 or P<0.05）.The changes of TGF-α in gastric tissue of rats with different polarities were not obvious.In terms of PGE₂,compared with the model group,5 flavor drugs were tested in two high-dose groups（petroleum ether and ethyl acetate sites）,3 flavor drugs were tested in the low dose group of petroleum ether（Alpinia officinarum Hance,Alpinia galanga Swartz,fructus galangae）,5 flavor drugs were tested in the low dose group of ethyl acetate,4 flavor drugs were tested in high dose group of n-butanol site（Alpinia officinarum Hance,Alpinia galanga Swartz,fructus galangae,Alpinia katsumadai）,in the low dose group of n-butanol site of fructus alpiniae galangae,3 flavor drugs were tested in the high dose group of water parts（fructus galangae,Alpinia katsumadai,Alpinia oxyphylla）all could enhance the expression of PGE₂ in gastric tissue,the difference was significantly obvious（P<0.01 or P<0.05）.The changes of PGE₂ the other polar parts in gastric tissue of rats were not obvious.In terms of EGF,compared with model group,4 different polar parts of the high dose group of 5 flavor drugs were tested,the low dose group of petroleum ether location of Alpinia katsumadai,the low dose group of ethyl acetate fraction of Alpinia galanga Swartz and fructus galangae,the low dose group of n-butanol site and water site of Alpinia galanga Swartz all could enhance the expression of EGF in gastric tissue,the difference was significantly obvious（P<0.01 or P<0.05）.The changes of EGF of the other polar parts in gastric tissue of rats were not obvious.（2）Effects of different polar parts of chinese herbal medicine on the contents of serum ET,NO,IL-8,TNF-α,VEGF,b FGF,PAF and NT in rats induced gastric ulcer and cold syndrome.In terms of ET,compared with model group,3 different polar parts of the high dose group of 5 flavor drugs were tested（petroleum ether,ethyl acetate and water）,the low dose group of petroleum ether location of Alpinia galanga Swartz and fructus alpiniae galangae,the low dose group of ethyl acetate fraction of 4 flavor drugs were tested（Alpinia galanga Swartz,fructus alpiniae galangae,Alpinia katsumadai,Alpinia oxyphylla）,4 flavor drugs were tested in the high dose group of n-butanol fraction and the low dose group of water site（Alpinia officinarum Hance,Alpinia galanga Swartz,fructus alpiniae galangae,Alpinia katsumadai）,the low dose group of n-butanol fraction of fructus alpiniae galangae all coμLd significantly reduce the expression of ET in serum of rats（P < 0.01 or P < 0.05）,the rest of the polarity parts each dose group of rats serum ET value change is not obvious.In terms of NO,compared with model group,2 different polar parts of the high dose group of 5 flavor drugs were tested（petroleum ether and water）,4 flavor drugs were tested in the low dose group of petroleum ether location（Alpinia officinarum Hance,Alpinia galanga Swartz,fructus alpiniae galangae,Alpinia katsumadai）,the high and low dose group of ethyl acetate fraction of 5 flavor drugs were tested,4 flavor drugs were tested in the high dose group of n-butanol fraction（Alpinia officinarum Hance,Alpinia galanga Swartz,fructus alpiniae galangae,Alpinia oxyphylla）,the low dose group of n-butanol fraction of Alpinia galanga Swartz,the low dose group of water site of Alpinia galanga Swartz and fructus alpiniae galangae all could significantly enhance the expression of NO in serum of rats（P < 0.01 or P < 0.05）,the rest of the polarity parts each dose group of rats serum NO value change is not obvious.In terms of IL-8,compared with model group,except the low dose group of petroleum ether part of fructus alpiniae galangae,the rest of the petroleum ether part high dosage group,low dose group of rats serum IL-8 value significantly decreased（P < 0.01 or P < 0.05）;five flavor drugs were tested of ethyl acetate part and n-butanol part of high and low dose group of rats serum IL-8 value significantly decreased（P < 0.01 or P < 0.05）;high dose group of water area of five flavor drugs were tested IL-8 value in rats serum were significantly lower（P < 0.01）;the rest of the polarity parts each dose group of rats serum IL-8 value changes are not obvious.In terms of TNF-α,compared with model group,2 different polar parts of the high dose group of 5 flavor drugs were tested（petroleum et-her and ethyl acetate parts）,the low dose group of ethyl acetate part of Alpinia officinarum Hance,3 flavor drugs were tested in the high dose group of n-butanol fraction（Alpinia galanga Swartz,fructus alpiniae gala-ngae,Alpinia katsumadai）,high dose group of water area of Alpinia o-xyphylla all could significantly reduce the expression of TNF-α in serum of rats（P<0.01 or P<0.05）,the rest of the polarity parts each dose group of rats serum TNF-α value change is not obvious.In terms of VEGF,compared with model group,2 different polar parts of the high dose group of 5 flavor drugs were tested（petroleum ether and ethyl acetate parts）,the low dose group of 5 flavor drugs were tested in petroleum ether and ethyl acetate parts,4 flavor drugs were tested in the high dose group of n-butanol fraction（Alpinia officinarum Hance,Alpinia galanga Swartz,fructus alpiniae galangae,Alpinia katsumadai）,4 flavor drugs were tested in the high dose group of water area（Alpinia officinarum Hance,Alpinia galanga Swartz,fructus alpiniae galangae,Alpinia oxyphylla）all could significantly reduce the expression of VEGF in serum of rats（P < 0.01 or P < 0.05）,the rest of the polarity parts each dose group of rats serum VEGF value change is not obvious.In terms of b FGF,compared with model group,high dose group of petroleum ether part and n-butanol part of 5 flavor drugs were tested,low dose group of petroleum ether part of Alpinia galanga Swartz and fructus alpiniae galangae,the high dose group of ethyl acetate part and water area of 5 flavor drugs were tested（Alpinia officinarum Hance,Alpinia galanga Swartz,fructus alpiniae galangae,Alpinia oxyphylla）,the low dose group of ethyl acetate part and water area of 2 flavor drugs were tested（Alpinia galanga Swartz and fructus alpiniae galangae）,the low dose group of n-butanol fraction of Alpinia officinarum Hance and fructus alpiniae galangae all could significantly reduce the expression of b FGF in serum of rats（P < 0.01 or P < 0.05）,the rest of the polarity parts each dose group of rats serum b FGF value change is not obvious.In terms of PAF,compared with model group,high and low dose group of 5 flavor drugs were tested in petroleum ether and ethyl acetate parts,high dose group of 5 flavor drugs were tested in n-butyl alcohol parts and water area,the low dose group of 3 flavor drugs were tested in n-butyl alcohol parts（Alpinia officinarum Hance,fructus alpiniae galangae,Alpinia oxyphylla）,the low dose group of 4 flavor drugs were tested in water place all could significantly reduce the expression of PAF in serum of rats（P < 0.01 or P < 0.05）,the rest of the polarity parts each dose group of rats serum PAF value change is not obvious.In terms of NT,compared with model group,the high dose group of 5 flavor drugs were tested in 4 polarity parts,the low dose group of 5 flavor drugs were tested in petroleum ether part,the low dose group of ethyl acetate parts of Alpinia officinarum Hance and Alpinia oxyphylla all could significantly reduce the expression of NT in serum of rats（P < 0.01 or P <0.05）,the rest of the polarity parts each dose group of rats serum NT value change is not obvious.Conclusion: 1.The drug substance basis of five flavour herbals of Alpinia treating rats gastric ulcer induced with cold syndrome are mostly concentrated in the petroleum ether and ethyl acetate parts,a small number parts of drug substance basis are concentrated in the n-butanol site and in the water.2.Rats induced gastric ulcer with cold syndrome are related to the drop of c AMP,PGE₂,EGF,TGF-alpha,NT and NO value;And the rise of IL-8,ET,TNF alpha,b FGF,VEGF and PAF values;Speculating the mechanism of effective parts of five flavour herbals of Alpinia treated ice acetic acid gastric ulcer with cold syndrome may be by reducing pepsin activity,improve the c AMP,PGE₂,EGF,TGF-alpha,NT,NO synthesis and release,inhibiting inflammatory cytokines IL-8 and TNF alpha,reduce ET,b FGF and VEGF,PAF values,playing warming stomach for dispelling cold,so as to repair the damaged gastric mucosa,in order to achieve the effect for the treatment of gastric ulcer cold syndrome.Indicating the relationship of traditional Chinese medicine is closer,its effectiveness is more approximate,the role of effective parts and pharmacodynamics effect are almost similar,its medicinal effect and chemical composition are related.