| The thesis consists of two parts,the first part is an animal experimental study;the second part is a literature review.In the literature review part,the etiology and pathogenesis of ankylosing spondylitis,Chinese medicine treatment and Western medicine treatment are discussed;the animal experimental study part explores the partial mechanism of action of long snake moxibustion on BALB/c mice induced ankylosing spondylitis from IL-23/Th17 axis.Research BackgroundAnkylosing Spondylitis(AS)is a chronic,progressive inflammatory disease with predominantly sacroiliac joint and spine involvement,starting with dull lumbosacral pain and morning stiffness in the early stages,which can be relieved by activity,but progressing to severe spinal ankylosis and severe joint dysfunction in the later stages.The cause of AS is still unclear and there is a lack of specific treatment.The incidence of AS in China is about0.3%.Although modern medical research on ankylosing spondylitis has been highly successful and there is a profound understanding of the pathogenesis and clinical features of this disease,it remains a common and difficult to treat disease.Current clinical treatment is focused on relieving pain,halting its progression,improving spinal mobility and maximizing the patient’s quality of life.In recent years,although there has been a great deal of research on the etiology,pathogenesis and treatment of ankylosing spondylitis,the use of Chinese medicine for the treatment of ankylosing spondylitis has made a large number of clinical reports,and the mentor group’s previous relevant clinical studies have also shown that the efficacy of long-snake moxibustion for the treatment of AS is certain,and long-snake moxibustion can not only significantly reduce or even eliminate the symptoms of AS patients,improve immune and inflammatory indicators to varying degrees,and slow down the development of the central axis and other The development process of the joints.However,few studies have been conducted on the mechanism of AS efficacy.What is the mechanism through which long snake moxibustion works in treating AS? Based on these facts,this study will explain the scientific implications of the mechanism of action of long snake moxibustion for AS from the perspective of AS and IL-23/Th17 immune inflammatory axis.We will explore the regulatory role of long snake moxibustion on IL-23/Th17 immune inflammatory axis and reveal some of the molecular mechanisms of this therapy in regulating the inflammatory response due to immune disorder of AS.Research ObjectivesIn this study,we constructed an animal model of ankylosing spondylitis(PGIA mouse model)to investigate the regulatory effect of long snake moxibustion on IL-23/Th17 immune inflammatory axis in ankylosing spondylitis mouse model,to reveal some of the molecular mechanisms of this therapy in suppressing inflammatory response and regulating immunity,and to provide theoretical basis for the study of the effect mechanism and clinical application of long snake moxibustion in treating ankylosing spondylitis.Research methodsIn the present study,32 female BALB/c mice were randomly generated into an empty control group,an AS-model group,a long snake moexib group and a celecoxib group,8 mice in each group except the empty group,the remaining 24 mice were generated using PGIA.BALB/c mice were injected intraperitoneally subcutaneously with bovine gastric proteoglycan to induce murine arthritis(PGIA)as an animal model of ankylosing spondylitis.Equal amounts of bovine proteoglycan were injected subcutaneously into the abdominal cavity of mice on days 0,20 and 40,respectively,and equal amounts of 0.9% saline were injected subcutaneously into the abdominal cavity of empty control mice.After a successful simulation,on days 41-70(1 month),the same amount of 0.9% saline as the celecoxib group was injected into the fasted control and model group mice,the long snake moccalysis group received long snake moccalysis and the celecoxib group received a clinically equivalent amount of celecoxib saline,and the above interventions were performed according to the experimental protocol [twice weekly for one month(8 times)].At the end of the experiment,the mice were treated and the experimental parameters were controlled and monitored.The main experimental parameters were:(i)hindfoot swelling in mice was assessed using the International Animal Arthritis Scale;(ii)serum was collected and expression levels of TNFa,IL-17 and IL-23 were determined by enzyme-linked immunosorbent assay(ELISA);(iii)swelling of the spine,sacrum and hindfoot of mice was monitored by DR;(iv)pathological tissue sections of the sacrum and hindfoot were stained with HE.Results1.Arthritis score: Compared with the blank control group,the arthritis and swelling scores of all groups of mice after PG moulding increased,and the difference was statistically significant(P<0.05),indicating that the moulding was successful,and after long snake moxibustion treatment,the arthritis scores decreased compared with the pre-treatment scores and the model group,and the difference was statistically significant(P<0.05),indicating that long snake moxibustion treatment may have an effect on PGIA mice by In addition,although both the long snake moxibustion and celecoxib groups were effective,there was no significant difference between the arthritis scores of the two groups,indicating that the difference between them was not statistically significant(P>0.05).2.serum TNF-α: Relative to the blank control group,serum TNF-α levels were significantly increased after PG modelling,with a statistically significant difference(P<0.05),indicating the presence of inflammatory responses and manifestation of IL-23/Th17 axis imbalance in the mice.After treatment with long snake moxibustion,the serum TNF-α of mice in the long snake moxibustion and celecoxib groups was significantly reduced compared with that in the model group,and the difference was highly statistically significant(P<0.01),indicating that under the intervention of long snake moxibustion,the inflammatory response of the body could be improved by reducing the concentration of TNF-α in the serum.In contrast,the serum TNF-α expression levels were similar in the long snake moxibustion group compared with the celecoxib group,indicating that the difference between them was not statistically significant(P>0.05).3.Serum IL-17,serum IL-23: Relative to the blank control group,the serum IL-17 and IL-23 levels were significantly increased after PG modelling,and the differences were all highly statistically significant(P<0.01),indicating the presence of inflammatory responses and manifestation of IL-23/Th17 axis imbalance in the mice.After treatment with long snake moxibustion,serum IL-17 was significantly lower in the long snake moxibustion and celecoxib groups compared with that in the model group,with a statistically significant difference(P<0.05);serum IL-23 was significantly lower in the long snake moxibustion and celecoxib groups compared with that in the model group,with a statistically significant difference(P<0.01),indicating that under the intervention of long snake moxibustion,serum IL-23 was significantly lower in the model group.It indicates that under the intervention of long snake moxibustion,the inflammatory response of the body can be improved by reducing the concentration of IL-17 and IL-23 in the serum.In contrast,serum IL-17 and IL-23 expression levels were similar in the long snake moxibustion group compared to the celecoxib group,indicating that the difference between the two was not statistically significant(P>0.05).4.Mice with swollen spine,sacroiliac joint and paw joint X-ray radiographs(DR): after modeling,compared with the blank control group,most mice showed different degrees of joint deformation and joint swelling in the paw joint and deformation in the sacroiliac joint,indicating successful modeling.5.HE-stained pathological tissue sections of sacroiliac and hind paw joints: In the blank control group,the margins of the sacroiliac and posterior claw joints were neat and the marginal cartilage was evenly distributed.In the model group,the sacroiliac and posterior claw joints had a large number of inflammatory cell infiltrations and chondrocyte hyperplasia,narrowing of the sacroiliac joint space,joint fusion,and gross and fibrotic sacroiliac joint margins.A small number of inflammatory cells were also seen in the sacroiliac and posterior claw joints of the long snake moxibustion and celecoxib groups,but the long snake moxibustion group had fewer irregularly deformed lymphocyte infiltrations,and the margins of the sacroiliac joint surfaces were neater and the articular fibrochondrocyte hyperplasia was not obvious,indicating that long snake moxibustion was more advantageous in improving joint bone destruction.Research ConclusionsThe animal model of AS(PGIA mouse model)has an imbalance of IL-23/Th17 inflammatory axis in vivo,and long snake moxibustion can exert immunomodulatory and anti-inflammatory effects by regulating IL-23/Th17 axis,and the immunomodulation and anti-inflammation are interrelated,which may be one of the mechanisms of long snake moxibustion for AS. |
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