| Purpose:To optimize the effect of Kuijie Prescription on HIF-2α pathway in UC spleen deficiency damp-heat syndrome and the expression of hypoxia-inducing factor HIF-2α in ulcerative colitis.Materials and METHODS: A total of 15 UC patients with spleen deficiency and dampness-heat syndrome admitted to the anorectal outpatient Department of the Third Affiliated Hospital of Liaoning University of Traditional Chinese Medicine and the Department of Traditional Chinese Medicine of Northern Theater General Hospital from November 2021 to July 2022 were enrolled as the UC treatment group.Kuijie prescription was optimized to intervene the patients with spleen deficiency and dampness-heat syndrome of UC,and was treated continuously for 4 weeks.Fifteen healthy subjects were recruited as healthy control group.Colonoscopy,modified Mayo score,endoscopic Mayo EMS score,C-reactive protein,erythrocyte deposition rate,intestinal mucosal biopsy HE staining,serum HIF-2α,IL-1β,IL-6 were detected in UC treatment group(before and after medication)and healthy control group.The expression of HIF-2α in intestinal mucosa was detected by IHC.The expression of HIF-2α mRNA in intestinal mucosa was detected by RT-qPCR.Serum HIF-2α expression level was correlated with improved Mayo score,CRP,ESR,inflammatory cytokines IL-1β and IL-6.Results:1.General information: There were 15 UC patients and 15 healthy patients in the study,and there were no significant differences in baseline data(gender,age and BMI)between the two groups(P > 0.05).2.Mayo score and EMS score: To optimize the prognosis of Rhizoma cuijiefang,there were statistical differences in the improvement of Mayo score and EMS score in UC treatment group(after medication)(P < 0.001),both of which were improved compared with before medication.3.CRP and ESR: The contents of CRP and ESR in UC treatment group(before medication)were significantly higher than those in healthy control group(P < 0.001),showing statistical difference;By optimizing the prognosis of kuijiefang Gan,CRP and ESR contents in UC treatment group were significantly decreased(P < 0.001),and no local or systemic adverse reactions were observed in UC treatment group.4.HE staining: In the healthy control group,the intestinal epithelial cells were arranged neatly,with normal shape and complete structure.The mucosa layer and mucosal muscle layer were evenly thin and thick,and no obvious inflammatory injury was observed.In the UC treatment group(before medication),there was damage and loss of intestinal mucosal epithelium,and a large number of inflammatory cells infiltrated with bleeding in the mucosa lamina propria.The state of intestinal mucosal inflammatory cell infiltration and bleeding in UC treatment group was improved compared with that before treatment.5.ELISA results: Serum HIF-2α,IL-1βand IL-6 contents in UC treatment group(before medication)were significantly higher than those in healthy control group(P < 0.001);The contents of HIF-2α,IL-1β and IL-6 in serum of UC treatment group(after treatment)were significantly lower than those before treatment(P < 0.01).6.IHC detection results: The expression of HIF-2α in intestinal mucosal tissues of UC treatment group(before medication)was significantly higher than that of healthy control group(P < 0.01);After treatment with optimized Kuijie formula,the expression of HIF-2α in intestinal mucosa of UC treatment group was significantly decreased(P < 0.01).7.RT-qPCR results showed that the mRNA expression of HIF-2α in intestinal mucosal tissues of UC treatment group(before medication)was significantly higher than that of healthy control group(P < 0.001);The mRNA expression of HIF-2α in intestinal mucosa was significantly decreased after treatment with the optimized Kuijie formula(P < 0.001).8.Correlation analysis: Serum HIF-2α expression level was positively correlated with the expression levels of modified Mayo score,CRP,ESR,inflammatory cytokines IL-1 β and IL-6(r=0.801,0.812,0.710,0.723,0.742,P < 0.001).Conclusions:1.The content of HIF-2α in serum and intestinal mucosal tissue of UC treatment group(before medication)was significantly higher than that of UC treatment group(after medication)and healthy control group,indicating that hypoxia indeed exists in UC and is related to the degree of disease.HIF-2α may play an important regulatory role in the pathogenesis of UC.2.Optimized Kuijie formula has a significant therapeutic effect on UC patients with spleen deficiency and dampness heat,and its mechanism of action may be related to HIF-2α pathway in many aspects.With the remission of the severity of the disease,the expressions of IL-1β,IL-6,CRP and ESR in serum of UC patients decreased significantly,and HIF-2α expression was positively correlated with the content of the above inflammatory factors.These results suggest that HIF-2α may affect the progression of UC disease through immune response.It is speculated that the optimized Kuijie formula may improve the changes of hypoxic microenvironment under the condition of intestinal mucosal inflammation by down-regulating the expressions of HIF-2α,IL-1 β and IL-6,relieve the clinical symptoms,relieve the inflammatory response,and promote the repair and healing of intestinal mucosa. |
