Study On Effective Ingredients Of Chinese Medicine Based On Traditional Chinese Medicine Prescription Compatibility Mechanism Reverse Esophageal Precancerous Lesions Induced By4-nitroquinoline-1-oxide In Mice

Objective:Establish an animal model of esophageal precancerous lesions induced bychemical carcinogen4-nitro-quinoline-1-oxide(4NQO) in C57BL/6mice.Observe the reversal effect of “Lian hua shen jia”formula made byForsythia Lignans, Trichosanthin, Cortex Periplocae triterpenoids anddangshen glycoprotein on various stages of esophageal precancerouslesions.Explore the possible mechanism of “Lian hua shen jia” formulareversing esophageal precancerous lesions.Methods:1150male and150female C57BL/6mice were randomly divided into sixgroups,Group A1(normal control group),Group A2(methyl cellulose controlgroup),Group B(simple induced cancer group),Group C(treatmentgroup),Group D(prevention group) and Group E(all-trans-retinoicacid,ATRApositive control group).GroupA1and A2each had30mice, group B,C,D andE had60mice in each group. Dissolved4NQO with sterilizing water to aconcentration of100μg/ml,given to B,C,D,E four groups of mice through thenormal drinking action method.“Lian hua shen jia”formula was made byForsythia Lignans: Trichosanthin: Cortex Periplocae triterpenoids: dangshenglycoprotein according to2mg:1.4mg:1.4mg:0.7mg mixing proportionsdissolved in1.5%methyl cellulose to a concentration of6.25percent.Group Dmice were administrated with “Lian hua shen jia”formula by gastric gavagefrom the beginning of the experiment. Group C mice were administrated with“Lian hua shen jia”formula by gastric gavage at15th week after simpleinduced cancer group B mice appeared atypical hyperplasia, Group E mice were administrated with “all-trans retinoic acid”(ATRA) by gastric gavage atthe same time with group C.Group C,D,E mice were fed by intragastricadministration,0.1ml/time, three times a week.2At12th and15th week, group A1,A2,B and D were killed five mice eachgroup to observe whether esophageal dysplasia appeared. At15th week groupsB,C,DandE mice were fed unifiedly by sterile water inatead of100μg/mlconcentration4NQO solution.At18th and21th week, group A1and A2werekilled five mice each group, group B,C,DandE were killed fifteen mice eachgroup.All the remaining mice in each group were killed at24th week.3Observed the reverse effect of “Lian hua shen jia”formula on esophagealprecancerous lesion of the mice models induced by4NQO with lightmicroscope and transmission electron microscope (TEM). Assessed the lesionseverity of mice’s esophageal tissue in each administration group and controlgroup by HE staining.Research the expression level of B7-H4molecule in theprocess of esophageal precancerous lesions in mice by immunohistochemicalanalysis. Compared the differential expressions of B7-H4molecules in eachadministration group and control group to evaluate the effect of B7-H4in theprocess of esophageal precancerous lesions.Results:1The progression of esophageal lesions in group C and group D wereslower than group B in the same period of the eighteenth,twenty-first andtwenty-fourth week,P＞0.05. There were no significant statistical differencesamong group B,C,D,E in the eighteenth and twenty-first week,but there weresignificant differences at twenty-fourth week, P<0.05. There were significantstatistical differences pairwise comparison between group B and C,group Band D, group B and E at twenty-fourth week, P<0.05. There were nosignificant statistical differences pairwise comparison between group C and D,groupC and E, group D and E at the same time.Showed that at twenty-fourthweek“Lian hua shen jia”formula can slow down the progression of esophageallesions by prevention or treatment,“Lian hua shen jia”formula had the samepharmacodynamic effect as ATRA at the sametime. 2In the twenty-first week,transmission electron microscope observed resultshowed that the esophageal tissue ultrastructure of group A1,A2,C,D allconnected tighter than that of group B, the intercellular space reduced.3In the process of4NQO inducing esophageal precancerous lesions ofmice,including mild dysplasia-moderate dysplasia-severe dysplasia-esophaguscancer,the B7-H4expression level showed significant statistical differences,P<0.05. And with the increased severity of esophageal precancerous,expression of B7-H4was gradually enhanced too,Suggesting that B7-H4molecules may play an important role in the progression of esophagealcarcinoma.4In the eighteenth and twenty-first week,there were no significantstatistical differences in B7-H4expression levels of mice esophageal tissueamong group B,C,D and E,P＞0.05,but in the twenty-fourth week, B7-H4expression levels in mice esophageal tissue among group B,C,D and E showedsignificant statistical differences, P<0.05.Showed that at the twenty-fourthweek, B7-H4expression levels of mice esophageal tissue in“Lian hua shenjia”formula prevention or treatment group were differentially expressedcompared with that in simple induced cancer group.Conclusion:14NQO induced the animal model of mice esophageal precancerous lesionssuccessfully.2The treatment or prevention with“Lian hua shen jia”formula can slowdown the procession of esophageal precancerous lesions development.3The B7-H4molecules may play an important role in the development ofesophageal carcinoma.4The effect of “Lian hua shen jia” formula may be slow down theprocession of4NQO induced mice esophageal precancerous lesions development by inhibiting the expression of B7-H4in esophageal tissue.