Abstract:

Objective

To investigate the effects of luole weimi ointment on oxidative stress and inflammatory cytokines expression in human esophageal epithelial cells （HEEC） induced by acid stimulation.

Methods

HEEC cells were cultured and the experiment was further divided into 5 groups：Control group（n=6）, acid stimulation group（n=6）, luoliwei pretreatment group （LL group, n=6）, pingkui jiawarixi maierwayite honey ointment pretreatment group （PK group, n=6） and esomeprazole pretreatment group（AS group, n=6）. The acid was stimulated with pH value acid medium （pH=4） 3 times a day, 15min each time, for 48 h； LL group, PK group and AS group were added with 1 mmol/L LL, PK and AS for 4 h, and then were stimulated with acid for 48 h. Total ribonucleic acid （RNA） was extracted from each group, and quantitative real-time polymerase chain reaction （RT-PCR） was performed to detect the mRNA expression levels of nicotinamide adenine dinucleotide phosphate oxidase-4 （Nox-4）, antioxidant enzymes ［manganese superoxide dismutase （Mn SOD）, glutathione peroxidase （GSH-Px）, catalase （CAT）］ and inflammatory factor indicators ［monocyte chemotactic protein 1 （MCP-1）, interleukin-6 （IL-6） and tumor necrosis factor α （TNF-α）］.

Results

After pH=4 acid stimulation for 48 h, the mRNA expression level of Nox-4 in HEEC cells was significantly increased （P＜0.0001）； the high expression of Nox-4 mRNA was inhibited in LL, PK and AS group （P＜0.0001）； the effect of LL group on inhibiting Nox-4 mRNA was better than that of PK group （P＜0.01）. pH=4 acid stimulation significantly reduced the expression of antioxidant enzymes （MnSOD,GSH-Px, CAT） mRNA in HEEC cells （P＜0.0001）. The mRNA expression levels of the above antioxidant enzymes were significantly increased in LL group, PK group and AS group （P＜0.0001）； compared with the PK group, the mRNA expression level of the antioxidant enzymes in LL group was significantly increased（P＜0.05）. Acid stimulation significantly increased the mRNA expression levels of inflammatory factors（MCP-1, IL-6, TNF-α） in HEEC cells （P＜0.0001）. The mRNA expression levels of the above inflammatory factors were significantly decreased in LL, PK and AS group （P＜0.0001）； it is noteworthy that compared with the PK group, the mRNA expression level of the inflammatory factors in LL group was significantly decreased （P＜0.01）.

Conclusion

Luole weimi ointment can significantly inhibit the oxidative stress and the inflammatory cytokines in HEEC cells induced by acid stimulation, and its mechanism needs further study.

Key words: Human esophageal epithelial cells, Luole weimi ointment, Acid stimulation, Oxidative stress, Inflammatory cytokines