| This paper is divided into two parts: The first part is animal experimental research and the second part is literature review.In the part of animal experiments,Part of the mechanism of long snake moxibustion on intestinal immunity and inflammation in mice with ankylosing spondylitis was discussed.The literature review part discusses the understanding and pathogenesis of ankylosing spondylitis from TCM/Western medicine,as well as treatment of TCM/Western medicine,animal model and other aspects.Research Background:Ankylosing spondylitis(AS)is one of the most intractable immune system diseases in the world,which has a long course of disease and is prone to recurrent attacks,and it is mainly in young and middle-aged males.It requires long-term treatment and has no specific drug or cure method.With the progression of ankylosing spondylitis patients may eventually lead to joint deformity and lifelong disability,which will not only have a great impact on the physical and mental health of AS patients,but also cause certain living pressure and economic burden to their families and society,which has become a hot issue of public health concern.In recent years,a large number of studies have been conducted on the etiology,pathogenesis and treatment of ankylosing spondylitis.A large number of clinical reports have been made on the treatment of ankylosing spondylitis with traditional Chinese medicine,which gradually deepens the understanding of ankylosing spondylitis and provides more effective methods and evidence support for clinical treatment of AS.However,the pathogenic factors,pathogenesis and therapeutic mechanism of ankylosing spondylitis have not been fully elucidated,especially there are few studies on the therapeutic mechanism.Previous related clinical studies of the tutor group have shown that in addition to having a good regulation effect on inflammation and immunity of AS,the long snake moxibustion has a significant effect on gastroenteritis symptoms such AS abdominal pain and diarrhea,which gives us a clue: In addition to regulating the immune function and improving the inflammatory state of AS patients,is the therapeutic effect of long snake moxibustion also related to the targeted regulation of intestinal immunity and inflammatory reaction of AS patients? If so,what is the mechanism of action? Based on the above background,this study systematically evaluated part of the mechanism of action of long snake moxibustion in the treatment of intestinal immunity and inflammatory response of ankylosing spondylitis through rigorous animal experimental design.This experiment comes from the National Natural Science Foundation project : “To investigate the mechanism of long snake moxibustion intervention in ankylosing spondylitis based on IL-23/Th17 immunoinflammatory axis”.Research Objective:In this paper,through the establishment of ankylosing spondylitis mice animal model,to explore the regulation effect of long snake moxibustion on intestinal immune and inflammatory response of mice model of ankylosing spondylitis,suggesting that part of the molecular mechanism of this therapy in regulating immune dysfunction and inhibiting inflammatory response in mice model of ankylosing spondylitis then providing theoretical basis for the study and clinical application of the effect mechanism.Research methods:On the basis of previous studies that have proved that long snake moxibustion has a good effect on ankylosing spondylitis,a mouse model of Proteoglycan Aggrecan-Induced Arthritis(PGIA)was induced by subcutaneous injection of bovine cartilage-derived proteoglycans into the intraperitoneal of mice,which was used as an animal model of ankylosing spondylitis.Thirty-two female BALB/c mice were randomly divided into four groups(8 mice in each group),which were blank control group,model group,long snake moxibustion group and sulfasalazine group.Except the blank control group,the other 24 mice underwent PGIA modeling.On day 0,day 20 and day 40,mice in the model group,long snake moxibustion group and sulfasalazine group were injected with the same amount of bovine cartilage-derived proteoglycans subcutaneously in the intraperitoneal,while mice in the blank control group were injected with 0.9%normal saline subcutaneously in the intraperitoneal.After the completion of modeling,during the 41-70 days(30 days)of the experiment,mice in the long snake moxibustion group were given long snake moxibustion treatment(twice a week),and clinical equivalent dose of sulfasalazine solution was given in the sulfasalazine group(once a day),and the blank control group and model group were given 0.9% normal saline gavage equal to sulfasalazine group(once a day).The above intervention measures were carried out according to the requirements of the experimental design.After the end of the experiment,the mice were treated with detection and observation in various relevant experimental indexes.The main indexes of this study were as follows: The international animal arthritis evaluation standard was used to score arthritis in mice;Spleen index(SI)and thymus index(TI)were measured;The expression levels of TNF-α,IL-17 and IL-23 in serum were determined by enzyme-linked immunosorbent assay(ELISA);The expression levels of IL-17 and IL-23 in ileum were detected by Western Blot;Histopathological observation of sacroiliac joint and posterior claw joint with HE staining.Research results:1.Arthritis score: Intra-group comparison: After treatment,the arthritis score of the long snake moxibustion group and the sulfasalazine group were significantly decreased compared with that before treatment,with highly significant statistical significance(P<0.01).Comparison among groups: After modeling,compared with blank control group,arthritis scores of model group,long snake moxibustion group and sulfasalazine group were significantly increased,with highly significant statistical significance(P<0.01);After modeling,there was no significant difference in arthritis score among model group,long snake moxibustion group and sulfasalazine group(P>0.05).After treatment,compared with model group,arthritis scores of long snake moxibustion group and sulfasalazine group were significantly decreased,with statistically significant differences(P<0.05).After treatment,there was no significant difference in arthritis scores between the long snake moxibustion group and sulfasalazine group(P>0.05).2.Spleen index(SI)and thymus index(TI): Comparison between groups: After the intervention of long snake moxibustion,compared with blank control group,SI of model group,long snake moxibustion group and sulfasalazine group were significantly increased,the differences were statistically significance(P<0.05);After the intervention of long snake moxibustion,SI of long snake moxibustion group was significantly lower than that of model group,and the difference was highly significant statistical significance(P<0.01).After the intervention of sulfasalazine,SI of sulfasalazine group showed a decreasing trend compared with model group,but there was no statistically significant difference(P>0.05).There was no significant difference in SI between Long snake moxibustion group and sulfasalazine group(P>0.05).Compared with blank control group,TI in model group was significantly increased,and the difference was highly significant statistical significance(P<0.01),while there was no significant difference in TI of long snake moxibustion group and sulfasalazine group compared with blank control group(P>0.05).After treatment,TI in the long snake moxibustion group and sulfasalazine group was significantly lower than that in the model group,with highly significant statistical significance(P<0.01).There was no significant difference in TI between the long snake moxibustion group and sulfasalazine group,with no statistical significance(P>0.05).3.Serum TNF-α: Comparison between groups: After the intervention of long snake moxibustion,compared with blank control group,serum TNF-α in model group,long snake moxibustion group and sulfasalazine group was significantly increased,and the differences were statistically significant(P<0.05);Compared with model group,serum TNF-α in long snake moxibustion group and sulfasalazine group were significantly decreased,with highly significant statistical significance(P<0.01).There was no significant difference in serum TNF-α between long snake moxibustion group and sulfasalazine group(P>0.05).4.Serum IL-17 and serum IL-23: Comparison between groups: After the intervention of long snake moxibustion,compared with blank control group,serum IL-17 and serum IL-23 in model group,long snake moxibustion group and sulfasalazine group were significantly increased,with highly significant statistical significance(P<0.01);Compared with model group,serum IL-17 in long snake moxibustion group and sulfasalazine group was significantly decreased,the difference was highly significant statistically significance(P<0.01),serum IL-23 in long snake moxibustion group and sulfasalazine group was significantly decreased,the difference was statistically significant(P<0.05).There was no significant difference in serum IL-17 and IL-23 expression between long snake moxibustion and sulfasalazine(P>0.05).5.IL-17 and IL-23 in ileum: After long snake moxibustion and other interventions,the histone expression levels of IL-17 and IL-23 in ileum of model group were significantly increased compared with blank control group;The histone expression levels of IL-17 and IL-23 in ileum of long snake moxibustion and sulfasalazine groups were increased compared with blank control group,but lower than that of model group,while the histone expression levels of IL-17 and IL-23 in ileum of long snake moxibustion group were lower than that of sulfasalazine group.6.HE stained pathological tissue sections of sacroiliac joint and posterior claw joint: After long snake moxibustion and other interventions,compared with the blank control group,the surface edge of the sacroiliac joint appeared coarse erosion,joint fibrochondrocytes proliferation,cartilage tissue degeneration and partial fibrosis,and inflammatory cells infiltration increased in the model group.In the model group,there were more inflammatory cells infiltration(deformed irregular lymphocytes)around the joint of the hind paw,erosion and destruction of the articular cartilage surface,and narrowing of the joint space.Compared with the model group,the damage and erosion of the sacroiliac joint surface margin in the long snake moxibustion group and the sulfasalazine group were less,the fibrous tissue hyperplasia was not obvious,and the inflammatory cells infiltration was reduced.Moreover,the improvement of the sacroiliac joint surface erosion,fibrous tissue hyperplasia and inflammatory cells infiltration in the long snake moxibustion group was more obvious than that in the sulfasalazine group.The inflammatory cells infiltration around the hind paw joint in the long snake moxibustion group and sulfasalazine group were less than that in the model group,and the inflammatory cells infiltration in the long snake moxibustion group was less than that in the sulfasalazine group.In addition,the joint cartilage surface erosion and joint space narrowing were improved in the long snake moxibustion group and sulfasalazine group compared with the model group,and the improvement of joint cartilage surface erosion and joint space narrowing in the snake moxibustion group was more obvious than that in sulfasalazine group.Research conclusions:Intestinal immune imbalance and inflammatory reaction were found in AS animal model(PGIA mice).Long snake moxibustion can inhibit the histone expression levels of IL-17 and IL-23 in ileum,and inhibit immune hyperfunction of SI and TI,moreover regulate cellular immune disorders and inflammatory responses caused by such disorders(TNF-α and inflammatory cells infiltration).Thereby playing a therapeutic role in intestinal immune imbalance and inflammatory reaction of ankylosing spondylitis,which may be one of the mechanisms of long snake moxibustion in the treatment of AS. |
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