A novel method for establishing a guinea pig model of reflux esophagitis
Objective To explore the effectiveness of establishing a Reflux Esophagitis(RE)mod-el in guinea pigs by ingesting pepsin-containing hydrochloric acid(pH=3)for different durations,determi-ning an optimal duration for a novel modeling approach.Methods A total of 40 male SD guinea pigs aged 3-4 weeks(purchased from Hubei Yizhicheng Biotechnology Co.,Ltd.)were randomly divided into con-trol,14-day model,21-day model,28-day model,and 35-day model groups,with 8 guinea pigs in each group.After 7 days of adaptation,modeling was initiated.The control group received sterile water(500 ml/day)for 28 consecutive days.The model groups were administered 0.1 mol/L hydrochloric acid containing 0.5％pepsin,with a pH of 3,at 500 ml/day,respectively for 14,21,28,and 35 days,re-spectively.General conditions and weight changes of the guinea pigs in each group were observed daily.After sample collection,the macroscopic appearance of the esophageal tissue was recorded,and histopatho- logical observations were conducted using hematoxylin-eosin(HE)staining.Quantitative real-time polymer-ase chain reaction(RT-PCR)was employed to assess the relative mRNA expression levels of interleukin-6(IL-6),interleukin-10(IL-10),tumor necrosis factor-α(TNF-α),Zonulaoccludens-1(ZO-1),and oc-cludin in the esophageal tissues of the guinea pigs.Additionally,protein expression of ZO-1 and occludin was evaluated using Western blotting.Comparison between the two groups was performed by t test.Results Compared to the control group,the activity of guinea pigs in each model group decreased,and their weight gain was slow,with some experiencing diarrhea and rash.Macroscopic observations revealed varying de-grees of congestion in the esophageal tissues of guinea pigs in each model group,with no apparent ulcera-tion.Under the microscope,early inflammatory damage to the esophageal mucosa was observed in all model groups.The 28-day and 35-day model groups exhibited noticeable thickening of the esophageal mucosa,in-flammatory cell infiltration,excessive proliferation of squamous epithelial cells,and elongation of the papil-lae in the submucosa,indicative of esophagitis.With prolonged modeling time,the expression of inflamma-tory factors increased,and the mucosal barrier function was compromised in the esophageal tissues of the model groups.Western blotting results showed that the protein expression levels of the tight junction protein ZO-1 in the 21-day,28-day,and 35-day model groups were lower than those in the control group(0.612± 0.051 vs.1.000±0.157,0.508±0.068 vs.1.000±0.157,0.514±0.094 vs.1.000±0.157,t=4.076,4.979,4.597,P＜0.05).Similarly,the protein expression levels of the tight junction protein Oc-cludin in the 14-day,21-day,28-day,and 35-day model groups were lower than those in the control group(0.758±0.120 vs.1.000±0.089,0.725±0.093 vs.1.000±0.089,0.444±0.054 vs.1.000± 0.089,0.443±0.099 vs.1.000±0.089,t=2.802,3.699,9.259,7.239,P＜0.05).The RT-PCR results indicated that the mRNA expression levels of ZO-1 in the 28-day and 35-day model groups were low-er than those in the control group(0.115±0.066 vs.1.208±0.796,0.130±0.038 vs.1.208±0.796,t=4.105,4.056,P＜0.05).Similarly,the mRNA expression levels of Occludin in the 14-day,21-day,28-day,and 35-day model groups were lower than those in the control group(0.679±0.148 vs.0.978± 0.098,0.551±0.227 vs.0.978±0.098,0.242±0.167 vs.0.978±0.098,0.126±0.103 vs.0.978±0.098,t=4.837,4.884,11.196,16.894,P＜0.05).Moreover,the mRNA expression levels of the inflammatory factor IL-6 in the 28-day and 35-day model groups were higher than those in the control group(10.985±5.300 vs.1.033±0.284,17.192±11.321 vs.1.033±0.284,t=-5.625,-4.281,P＜0.05).Similarly,the mRNA expression levels of the inflammatory factor TNF-α in the 28-day and 35-day model groups were higher than those in the control group(7.565±2.729 vs.1.014± 0.182,13.190±5.530 vs.1.014±0.182,t=-5.449,-4.92,P＜0.05).Additionally,the mRNA expression levels of the anti-inflammatory factor IL-10 in the 14-day,21-day,28-day,and 35-day model groups were lower than those in the control group(0.486±0.273 vs.1.010±0.150,0.309±0.206 vs.1.010±0.150,0.190±0.171 vs.1.010±0.150,0.215±0.126 vs.1.010±0.150,t=5.043,8.253,10.822,12.203,P＜0.05).Conclusion A guinea pig model of reflux esophagitis can be suc-cessfully established through self-administration of acid.The optimal modeling duration is 28 days,laying the foundation for subsequent research.
Reflux esophagitisSelf-administration of acidGuinea pigsAnimal model
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