Molecular Mechanism Of Sinomenine Hydrochloride Mediated Notch Signaling Pathway In The Treatment Of Ulcerative Colitis In Mice

Objective: To study the therapeutic effect of sinomenine hydrochloride（SH）on ulcerative colitis（UC）in mice as an animal model and the changes of Notch signaling pathway in colon tissue of mice after treatment.Methods: The experiment set up four groups: normal control group,model group,SH low-dose treatment group（20 mg/kg）,and SH high-dose treatment group（60 mg/kg）,with 6 BALB/c mice in each group.The experimental mice established UC model by drinking 4% dextran sodium sulfate（DSS）aqueous solution freely for 7 days.The normal control group drank purified water freely and was gavaged with purified water for 7 days;the model group drank 4% DSS aqueous solution freely and gavaged with purified water for 7 days;SH low-dose and high-dose treatment groups freely drank 4% DSS aqueous solution while ad libitum.SH 20mg/kg and SH 60mg/kg were given by gavage for 7 days respectively.The disease activity index（DAI）,colon length,colon histological injury pathology score of mice in each group were recorded and compared;q RT-PCR was used to detect the gene expression level of Notch signaling pathway Notch1 receptor in colon specimens of mice;The expression levels of Notch signaling pathway Notch1,NICD1,Jagged1,and Hes1 proteins were detected by western blot;the expression levels and expression sites of Notch signaling pathway Notch1,NICD1,Jagged1,and Hes1 proteins were detected by immunohistochemistry;ELISA was used to detect the cytokine expression levels of TNF-α,IL-1β,and IL-6.Results: Compared with the normal control group,the DAI of the mice in the model group was significantly increased（P<0.001）,the length of the colon was significantly shortened（P<0.001）,the pathological score of colon histological damage was significantly increased（P<0.001）,the expression of Notch1 receptor gene in colon tissue was significantly increased（P<0.001）,and Notch1,NICD1,Jagged1,Hes1 protein expression levels in the colon tissue were significantly increased（P<0.001）,and serum cytokines TNF-α,IL-1β,IL-6 expression levels were also significantly increased（P<0.001）;Compared with the model group,the SH treatment group（low and high doses）showed a significant decrease in DAI（P<0.001）,colon shortening was significantly improved（P<0.05）,and the pathological score of colon histological damage was significantly decreased（P<0.01）,the expression of Notch1 receptor gene in colon tissue was significantly decreased（P<0.001）,the protein expression levels of Notch1,NICD1,Jagged1,and Hes1 in colon tissue were significantly decreased（P<0.01）,and Notch1 and Jagged1 were mostly expressed in the cell membrane,while NICD1 and Hes1 were mostly expressed in the nucleus.The expression levels of TNF-α,IL-1β and IL-6 were also significantly decreased（P<0.001）.Conclusion: SH has a protective effect on DSS-induced UC in mice,and the mechanism may be related to its inhibition of the abnormal activation of Notch signaling pathway in colon tissue and the inhibition of the expression of pro-inflammatory cytokines TNF-α,IL-1β,and IL-6.