The MRNA Expressions Of NF-kappa B And Epigenetics-related Factors In GES-1 Cells Infected By Helicobacter Pylori Treated With Human Medicated Serum Of Banxia Xiexin Decoction

Objective:From the perspective of NF-?B signal pathway and epigenetic related factors,the possible mechanism of Banxia Xiexin Decoction in the treatment of peptic ulcer was studied by detecting the expression levels of NF-?B,NF-?Bp65,I?B?,Dnmt1,MBD2 and kaiso mRNA after the intervention of GES-1 cells in different periods,so as to enrich the scientific connotation of Banxia Xiexin Decoction in the treatment of peptic ulcer.Methods:1.Collection of clinical human drug-containing serum:clinical data sources were divided into three groups,namely,healthy control group,western medicine group and traditional Chinese medicine group.The health control group came from 40 healthy volunteers from the Health Examination Center of the Second People's Hospital Affiliated to Fujian University of Traditional Chinese Medicine,half male and half male,aged 20-50 years old.Eg drug treatment group for peptic ulcer came from patients with cold and heat mixed syndrome of Hp-related PU in Xiamen Traditional Chinese Medicine Hospital and National Medical Hall affiliated to Fujian Traditional Chinese Medicine.Each drug treatment group of peptic ulcer was randomly split into two groups,40 men and women,aged 20-50 years old.Sign informed consent.Western medicine group was to lansoprazole+clarithromycin+metronidazole,twice a day,oral;Chinese medicine group to Banxia Xiexin decoction,twice a day,oral.Intravenous blood was collected before treatment and 7 days after treatment,and serum was collected and separated for later use.2.Cell Intervention and Collection:GES-1 cells were cultured in a high glucose DMEM medium with penicillin,streptomycin and 10%fetal bovine serum,which was placed in a37?incubator and 5%CO₂.Use antibiotic-free high-sugar DMEM medium when mixed with Hp.After grouping,the corresponding experimental serum culture was given.GES-1culture medium without Hp was used as blank control group.In addition,the mixture of Hp and GES-1 was cultured in a 6-part culture box for 24 hours?starting from the addition of Hp?,1 part without serum intervention?that is,the Hp model group?,and the other 5 parts were added to the serum intervention:the healthy serum control group added the drug-free serum of the healthy patient;the pre-drug group added the pre-treatment serum of the patient;the western medicine serum group added the drug-containing serum of the western medicine group;and the serum concentrations of the intervention culture of the traditional Chinese medicine A and B groups were 10%and 20%,respectively.Each group collected cells after 1,2 and 3 days of continuous culture.3.RT-qPCR was used to detect the expression levels of NF-?B,NF-?Bp65,I?B?,Dnmt1,MBD2 and kaiso mRNA in GES-1 cells after 24h,48h and 72h.And draw their respective dissolution curves..Results:1.According to the PCR amplification dissolution curve,the dissolution temperatures of NF-?B and GAPDH were about 82,86and86,respectively,indicating that the PCR amplification product was a single product.The amplification efficiency of NF-?B and GAPDH is basically the same.2.RT-qPCR was used to detect the expression levels of,NF-?B NF-B?p65 and I?B?mRNA in each group after 24 hours,48 hours and 72 hours of GES-1 cells.The results showed that compared with the blank group,NF-?B NF-B?p65 and I?B?mRNA increased and I?B?mRNA decreased in the Hp infection model group?P<0.05?;compared with the Hp infection model group,the expression of NF-?B,NF-?Bp65 mRNA decreased in the western medicine serum group and Banxia Xiexin decoction serum group.Compared with the pre-medication serum group,the expression of NF-?B and NF-B?p65 mRNA in the western medicine serum group and 20%Banxia Xiexin Decoction serum group decreased,and the expression of I?B?mRNA increased?P<0.05?.There was no difference in the expression of mRNA between Banxia Xiexin Decoction serum group and western medicine serum group?P>0.05?.3.RT-qPCR was used to detect the expression of Dnmt1,MBD2 and kaiso mRNA in GES-1 cells after 24h,48h and 72h.The results showed that compared with the blank group,the expression of Dnmt1 and kaiso mRNA in the Hp infection model group was increased?P<0.05?;compared with the Hp infection model group,there was no difference in the expression of mRNA in the healthy serum group?P>0.05?.The expression of A was decreased?P<0.05?;compared with the pre-medication serum group,the expression of Dnmt1 and kaiso mRNA in the western medicine serum group and 20%Banxia Xiexin Decoction serum group was decreased?P<0.05?;there was no difference in the expression of mRNA between the Banxia Xiexin Decoction serum group and the western medicine serum group?P>0.05?,but there was no significant difference in the expression of MBD2 mRNA before and after medication?P>0.05?.Conclusion:1.The drug-containing serum of Banxia Xiexin Decoction could reduce the expression level of NF-?B and NF-?Bp65 mRNA in GES-1 cells and increase the expression level of I?B?mRNA.2.The human drug-containing serum of Banxia Xiexin Decoction could reduce the expression of Dnmt1 and kaiso mRNA in GES-1 cells.3.The mechanism of Banxia Xiexin Decoction in the treatment of peptic ulcer is related to the inhibition of pro-inflammatory activity by lowering the expression levels of NF-?B and NF-?Bp65,increasing the expression level of I?B?,reducing the phosphorylation degradation of I?B?,inhibiting the activation of NF-?B,regulating the signal pathway of NF-?B,and then reducing the inflammatory response of Hp-related PU.It may also be one of the mechanisms to participate in the regulation of cell DNA methylation by regulating or inhibiting the expression of Dnmt1 and kaiso mRNA.