Study On The Immune Regulation Mechanism Of Yukouning And Its Clearing Heat And Detoxifying Prescription In Rats With Recurrent Aphthalmus Of Heart-Spleen Accumulation Type

Background:Recurrent aphthous ulcer(RAU)is the most common oral mucosal disease in clinic.High incidence,high recurrence rate and obvious local pain are the main characteristics of the disease.At present,symptomatic treatment(local anti-inflammatory pain relief+systemic immune regulation)is the main treatment of RAU.Commonly used topical application or gargle such as aureomycin can produce side effects such as tooth staining,blackening of tongue fur and abnormal taste.Hormones and Immunoenhancers are not only proved to severe side effects such as Cortical hyperfunction and Osteoporosis,but also can not prevent the recurrence of this disease.Therefore,RAU is still one of the oral refractory diseases.It is important to actively develop new treatment methods for RAU patients.Object:To study the immunoregulatory mechanism of Yukouning and its decomposed prescription for clearing away heat and detoxifying toxins in rats with recurrent aphthous ulcer caused by accumulation of heat in the heart and spleen.Methods:40 RAU(Heart-Spleen Accumulation-Heat Syndrome)SD rats with oral ulcer were prepared by immune induction combined with model irritation and high-fat spicy diet.They were randomly divided into four groups,Yukouning,Yukouning’s prescription for clearing away heat and separated recipes,Levamisole Hydrochloride and normal saline were administered for 14 consecutive days.T lymphocyte subsets in peripheral blood of rats were detected and the pathological changes of ulcer mucosa were observed under light microscope.The oral ulcer model of 40 RAU(Heart-Spleen Accumulation Heat Syndrome)SD rats was prepared by immune induction combined with model irritation and high-fat spicy diet.The rats were randomly divided into four groups,Yukouning,Yukouning’s prescription for clearing away heat and separated recipes,Levamisole Hydrochloride and normal saline were given for 14 consecutive days.The levels of EGF and TGF-β were detected by ELISA and flow cytometry,respectively.The expressions of EPI3K/AKT signal transduction pathway proteins and related transcription factors in ulcer mucosa were detected by Western blot.According to the 40 cases of patients who were diagnosed as RUA(heart spleen and heat syndrome)from March 2017 to March 1818 in March,they were classified according to the order of the treatment,and were divided into odd and even groups according to odd and even numbers.Twenty patients in the levamisole group were treated with yoghurt and levamisole for 4 weeks.The clinical effects of the two groups were observed,and the blood values of CD4+,CD8+,CD4+/CD8+were observed.Results:Compared with normal saline group,Yukouning,Yukouning’s prescription for clearing away heat and separated recipes and Levamisole Hydrochloride could promote the healing of RAU ulcer.Flow cytometry was used to detect and compare the changes of T cell subsets in Normal saline group,Yukouning,Yukouning’s prescription for clearing away heat and separated recipes and Levamisole Hydrochloride.Compared with Normal saline group,the levels of CD4+and the ratio of CD4+/CD8+in Yukouning group,Yukouning’s prescription for clearing away heat and separated recipes group and Levamisole Hydrochloride group increased significantly(P<0.05),while the levels of CD8+decreased significantly(P<0.05).Compared with Normal saline group,the EGF levels in Yukouning group,Yukouning’s prescription for clearing away heat and separated recipes group and Levamisole Hydrochloride group were significantly higher(P<0.05),and the expression levels of PI3K,p-PI3K,Akt,p-Akt and NF-κB related genes in PI3K/AKT signaling pathway were increased.Research conclusions:Yukouning can regulate the immune function of RAU(Heart-Spleen Accumulation-Heat Syndrome),which may be an important mechanism of Yukouning in promoting ulcer healing and reducing recurrence.Yukouning and its prescription for clearing away heat and separated recipes can regulate EGF,activate PI3K/Akt signal transduction pathway,regulate BCR signal transduction,activate Bregs,regulate immune function,promote the healing of recurrent aphthous ulcer of the type of accumulation of heat in the heart and spleen,and reduce the recurrence of ulcer.