不同艾灸距离对Ⅱ型胶原诱导型关节炎大鼠疗效比较的实验研究

发布时间：2018-05-26 05:47

本文选题：距离 + 艾灸　；参考：《南京中医药大学》2017年硕士论文

【摘要】：背景类风湿关节炎(Rheumatoid Arthritis,RA)是一种原因不明的,以关节滑膜炎症为特征的慢性自身免疫性疾病,主要累及小关节[1]。RA会引起患者出现关节损伤,甚至最终导致残疾,降低了患者的生活质量,甚至可以引起心血管及其他合并症。改善病情的抗风湿药物(Disease-modifying antirheumatic drugs,DMARDs)为治疗 RA 的关键药物,其通过减少滑膜炎症和全身炎症从而改善关节功能。当关节炎不受控制或者DMARDs联用产生效应时,应使用生物制剂。但是产生感染、结核、肿瘤的危险以及昂贵的价格令患者难以承受,从而限制了其在临床上的使用。艾灸作为中医外治法的重要组成部分,可以用于治疗多种慢性疾病。利用艾叶燃烧而产生的温通温补作用及艾叶的药物作用,经过经络的调节,发挥补益气血阴阳,驱寒散湿的功效。艾灸能够温通经络,活血化瘀,不仅对炎症反应局部有抗炎调节的作用,而且能够通过调节神经-内分泌网络系统而对整个机体进行调控,从而达到治疗类风湿关节炎的目的。但是,艾灸操作大多凭借经验,临床缺乏标准,艾灸距离成为影响艾灸疗效的重要因素之一。目的通过比较不同距离艾灸治疗类风湿关节炎的疗效差异,探索治疗RA最佳的艾灸距离。为艾灸在临床的推广与应用提供科学依据,以及指导未来的科学实验及临床应用。方怯(1)选用SPF级雄性SD大鼠40只,随机抽取8只做为正常对照组,余32只全部建立胶原诱导性关节炎(Collagen-Induced Arthritis,CIA)大鼠模型。32只大鼠全部造模成功,按随机数字表法进行随机分组,分为模型组、艾灸1cm组、艾灸2cm组、艾灸3cm组,每组8只。正常对照组和模型组不予艾灸治疗,但给予相同的时间固定,每次40min。艾灸1cm组每天灸双侧"肾俞"和"足三里"穴,每天1次,每个穴位10min,共40min,保持灸条距穴位皮肤的距离维持在1cm。艾灸2cm组与艾灸3cm组分别保持灸条距离穴位皮肤为2cm、3cm,其余同艾灸1cm组。每6 d为1个疗程,共干预3个疗程,疗程间休息1d。(2)基线比较:比较造模前各组大鼠体质量、足趾容积。(3)观察各组大鼠造模前、干预前、艾灸3周后的一般情况,包括被毛色泽、摄食情况、四肢活动情况等。(4)测量并比较各组大鼠干预前、艾灸3周后的体质量变化。(5)比较各组大鼠干预前、艾灸3周足趾容积变化。(6)根据大鼠四肢的足趾肿胀及变形程度等来评定大鼠在干预前、艾灸3周的关节炎指数(Arthritis Index,AI)得分变化,正常对照组无此项得分。(7)艾灸3周后,腹主动脉采血、离心,进行酶联免疫吸附剂测定(Enzyme Linked ImmunosorbentAssay,ELISA),比较各组大鼠血清白细胞介素1β(IL-1β)、肿瘤坏死因子α(TNF-α)的水平。(8)艾灸3周后,收集各组大鼠一侧踝关节病理组织标本进行病理组织形态学观察。结果(1)基线比较:各组大鼠造模前体质量、足趾容积无统计学差异(P0.05),具有可比性。(2)一般情况:造模前,各组大鼠一般情况良好;造模后,模型组、艾灸各组大鼠均出现关节红肿,部分大鼠出现注射局部皮肤溃疡;给予艾灸治疗3周后,模型组大鼠症状持续加重,关节肿胀明显,不能负重,活动受限,且部分大鼠出现跛行;艾灸各组大鼠饮食尚可,活动较模型组更为活跃,关节肿胀明显改善,但仍有轻微关节红肿;正常对照组表现无异常。(3)体质量比较:造模前,各组大鼠体质量差异无统计学意义(P0.05),基线一致。造模后干预前,与正常对照组相比,模型组、艾灸各组体质量增长缓慢,差异有显著统计学意义(P0.01)。艾灸各组和模型组体质量比较,差异无统计学意义(P0.05)。艾灸3周后,正常对照组大鼠体质量持续增长明显;与正常对照组相比,模型组大鼠体质量增长缓慢,差异有显著统计学意义(P0.01);与模型组相比,艾灸3cm组体质量增长较快,差异具有统计学意义(P0.05);与模型组相比,艾灸1cm组、艾灸2cm组大鼠体质量增长明显,差异有显著统计学意义(p0.01);艾灸各组之间无统计学差异(p0.05)。(4)足趾容积比较:艾灸干预前,与正常对照组相比,模型组和艾灸各组大鼠足趾容积增加明显,P0.01,差异具有显著统计学意义;与模型组相比,艾灸各组足趾容积无统计学差异(P0.05),具有可比性。艾灸3周后,与正常对照组相比,模型组大鼠足趾容积增长明显,P0.01,差异具有显著统计学意义;与模型组相比,艾灸各组足趾容积明显下降,P0.01,差异具有显著统计学意义;与艾灸3cm组相比,艾灸1cm组与艾灸2cm组大鼠足趾容积下降明显,P0.01,差异具有显著统计学意义。艾灸1cm组与艾灸2cm组之间无统计学差异(P0.05)。(5)关节炎症评分比较:干预前,除正常对照组外,各组大鼠AI评分均大于4分,显示造模成功,组间比较,P0.05,差异无统计学意义,具有可比性。艾灸3周后,模型组AI评分持续增高,艾灸各组AI评分较模型组明显下降(P0.01)。艾灸各组之间比较,AI评分差异无统计学意义(P0.05)。(6)IL-1β水平:艾灸3周后,与正常对照组相比,模型组血清IL-1β水平明显升高,P0.01,差异有显著统计学意义。与模型组相比,艾灸各组血清IL-1β水平明显下降,P0.01,差异有显著统计学意义。与艾灸3cm组相比,艾灸1,2cm组血清IL-1β水平较明显降低(P0.01),艾灸1cm组与艾灸2cm组之间差异无统计学意义(P0.05)。(7)TNF-α水平:艾灸3周后,与正常对照组相比,模型组血清INF-α水平明显升高,P0.01,差异有显著统计学意义。与模型组相比,艾灸各组血清TNF-α水平明显下降,P0.01,差异有显著统计学意义。与艾灸3cm组相比,艾灸1,2cm组血清TNF-α水平较明显降低(P0.01),艾灸1cm组与艾灸2cm组之间差异无统计学意义(P0.05)。(8)病理HE染色:正常对照组大鼠踝关节病理HE染色未见明显异常。模型组可见关节腔变窄,狭窄的关节腔内可见炎细胞;炎细胞严重浸润,血管翳形成;滑膜细胞肿胀且排列无序,有明显增生,并且可见严重变性坏死。艾灸各组可见中度炎细胞浸润;滑膜细胞稍有增生或中度肿胀;与模型组比较,血管翳明显减少,表明具有明显好转。艾灸2cm组大鼠滑膜肿胀、炎细胞浸润程度较艾灸1cm组和艾灸3cm组有所改善,而艾灸1cm组与艾灸2cm组间无明显差异。结论(1)艾灸3周治疗后,与模型组相比,艾灸各组一般情况改善,足趾红肿相对较轻,病理染色显示滑膜变薄、炎细胞轻度浸润。血清IL-1β水平、TNF-α水平均低于模型组。证明艾灸在改善关节功能、调节免疫方面具有一定作用。(2)艾灸治疗CIA大鼠,在艾灸距离为1～2cm时,艾灸治疗效果更佳。(3)从耐受性、接受度、疗效等方面综合考虑,艾灸2cm组为艾灸治疗类风湿关节炎大鼠的最佳距离。本实验研究结果为临床规范化应用艾灸治疗RA提供重要的参考价值和借鉴意义。
[Abstract]:Background rheumatoid arthritis (Rheumatoid Arthritis, RA) is a chronic autoimmune disease characterized by joint synovitis. The main involvement of the small joint [1].RA can cause joint injury, even eventually lead to disability, reduce the quality of life of the patient, even cause cardiovascular and other complications. Disease-modifying antirheumatic drugs (DMARDs) is a key drug for the treatment of RA. It improves joint function by reducing synovitis and systemic inflammation. Biological agents should be used when arthritis is not controlled or DMARDs is produced. The expensive price makes the patient difficult to bear, which limits its clinical use. Moxibustion is an important part of the external treatment of traditional Chinese medicine. It can be used to treat a variety of chronic diseases. Effect. Moxibustion can warm the meridian, promote blood circulation and dissipate blood stasis, not only has the effect of inflammation and regulation of inflammation, but also can regulate the whole body by regulating the nervous endocrine network system, so as to achieve the purpose of treating rheumatoid arthritis. However, moxibustion is mostly operated by the experience, the clinical lack of standard, moxibustion distance becomes Objective one of the important factors affecting the therapeutic effect of moxibustion is to compare the difference of therapeutic effect of different distance moxibustion on the treatment of rheumatoid arthritis, explore the best distance of moxibustion for the treatment of RA, provide scientific basis for the popularization and application of Moxibustion in clinical practice, and guide the future scientific experiment and clinical application. (1) 40 male SD rats were selected. 8 rats were randomly selected as the normal control group, and the other 32 rats were all established with collagen induced arthritis (Collagen-Induced Arthritis, CIA) rat model. All the rats were successfully modeled. The random numbers were randomly divided into model group, moxibustion 1cm group, moxibustion 2cm group, moxibustion 3cm group, and group of moxibustion 3cm group, with 8 rats in each group. Moxibustion treatment, but given the same time fixed, each 40min. moxibustion 1cm group each day moxibustion bilateral "Shenshu" and "Zusanli" point, 1 times a day, each point 10min, a total of 40min, keep the distance between the moxibustion bar and the point skin on the 1cm. moxibustion 2cm group and moxibustion 3cm group to maintain the distance point skin for 2cm, 3cm, the rest of the moxibustion and 1cm group. Every 6 d. 1 courses of treatment, a total of 3 courses of intervention, interval of rest 1D. (2) baseline comparison: compare the mass of rats in each group before making the model and the volume of the toe. (3) observe the general condition of the moxibustion before 3 weeks of moxibustion before the intervention, including the hair color, feeding situation, and the living conditions of the limbs. (4) measure and compare the moxibustion for 3 weeks before the intervention of rats in each group. Change of body mass. (5) compare the volume changes of moxibustion for 3 weeks before intervention. (6) according to the toe swelling and deformation degree of the limbs of the rats, the scores of Arthritis Index (AI) of the moxibustion for 3 weeks before intervention were evaluated, and there was no score in the normal control group. (7) the abdominal aorta was collected and centrifuged after 3 weeks of moxibustion. Enzyme Linked ImmunosorbentAssay (ELISA) was used to compare the level of serum interleukin 1 beta (IL-1 beta) and tumor necrosis factor alpha (TNF- alpha) in all groups of rats. (8) after 3 weeks of moxibustion, pathological histological specimens of one side of the ankle joint were collected for pathological histomorphology. Results (1) baseline comparison: the model of rats in each group There was no statistical difference between the mass of the precursor and the volume of the toe (P0.05). (2) general situation: before the model, the rats in each group were in good condition. After the model, the model group and the moxibustion group all had joint redness and swelling, and some rats were injected with local skin ulcers. After the moxibustion treatment was given for 3 weeks, the symptoms of the rats in the model group continued to aggravate and joint swelling. Obviously, the weight was not heavy, the activity was limited, and some rats were limping; the rats in the moxibustion group were more active in diet, more active than the model group, and the joint swelling was obviously improved, but there was still slight joint swelling and swelling in the normal control group. (3) the body mass comparison: before the model, the body mass difference was not statistically significant (P0.05), baseline a Before the model intervention, compared with the normal control group, the body mass of the model group and moxibustion had a significant statistical significance (P0.01). There was no significant difference between moxibustion and model group body mass (P0.05). After 3 weeks of moxibustion, the body mass of the normal control group continued to increase obviously; compared with the normal control group, the model group was compared with the normal control group. The body mass of rats increased slowly, the difference was significant (P0.01). Compared with the model group, the body mass of moxibustion 3cm group increased rapidly, and the difference was statistically significant (P0.05). Compared with the model group, the body mass of moxibustion 1cm group and moxibustion 2cm group increased significantly (P0.01), and there was no statistical difference between moxibustion groups (P0.01); there was no statistical difference between moxibustion groups. (P0.05). (4) comparison of toe volume: before moxibustion, the volume of toe in model group and moxibustion group increased significantly compared with that of moxibustion group, P0.01, the difference had significant statistical significance. Compared with model group, there was no statistical difference between moxibustion and moxibustion in each group (P0.05). After moxibustion, moxibustion was compared with the normal control group, after 3 weeks, the model was compared with the normal control group. The volume growth of the toes in the group of rats was significantly increased, P0.01, the difference had significant statistical significance. Compared with the model group, the volume of the toe of moxibustion was significantly decreased, P0.01, the difference had significant statistical significance. Compared with the moxibustion 3cm group, the volume of the toe of Moxibustion 1cm group and moxibustion 2cm group decreased significantly, and the difference was significant statistically significant. Moxibustion 1cm was significant. There was no statistical difference between group and moxibustion 2cm group (P0.05). (5) comparison of arthritis score: before intervention, except normal control group, the AI score of rats in each group was more than 4 points, which showed that the model was successful, the difference between groups, P0.05, the difference was not statistically significant. After 3 weeks of moxibustion, the AI score of the model group continued to increase, and the AI score of moxibustion groups was more than that of the model group. There was no significant difference (P0.01). There was no significant difference in AI score between groups of Moxibustion (P0.05). (6) IL-1 beta level: after 3 weeks of moxibustion, the level of serum IL-1 beta in the model group was significantly higher than that of the normal control group, and the difference was statistically significant. Compared with the model group, the level of serum IL-1 beta in moxibustion groups decreased significantly, and P0.01, the difference was significant. Statistical significance. Compared with the moxibustion 3cm group, the serum IL-1 beta level of moxibustion 1,2cm group was significantly lower (P0.01), the difference between moxibustion 1cm group and moxibustion 2cm group was not statistically significant (P0.05). (7) the level of TNF- A: after 3 weeks of moxibustion, the level of serum INF- alpha in the model group was significantly higher than that in the normal control group, and the difference of P0.01, with significant statistical significance, was significant. Compared with the moxibustion group, the serum TNF- alpha level was significantly decreased, P0.01, the difference was significant statistically significant. Compared with the moxibustion 3cm group, the serum TNF- alpha level of the moxibustion 1,2cm group was significantly lower (P0.01), the difference between the moxibustion 1cm group and the moxibustion 2cm group was not statistically significant (P0.05). (8) pathological HE staining: pathological HE staining of the ankle joint in the normal control group. No obvious abnormality was found in the model group. Inflammatory cells were seen in the joint cavity; inflammatory cells were seriously infiltrated and pannus formed; the synovial cells were swollen and disordered, with obvious hyperplasia and severe degeneration and necrosis. The moxibustion groups showed moderate inflammatory cells in each group; the synovial cells were slightly proliferated or moderately swelled; and the model group was a model group. Compared with the moxibustion 2cm group, the synovial swelling of the moxibustion group was obviously better than that of moxibustion 1cm group and moxibustion 3cm group, but there was no significant difference between moxibustion 1cm group and moxibustion 2cm group. Conclusion (1) after moxibustion for 3 weeks, the moxibustion was compared with the model group, the general condition of moxibustion was improved and the toes red and swelling were relatively more than that of the model group. Light, pathological staining showed that the synovial membrane was thinner and the inflammatory cells were slightly infiltrated. The level of serum IL-1 beta and TNF- alpha water were lower than the model group. It was proved that moxibustion had a certain role in improving the function of joint and regulating immunity. (2) moxibustion was used to treat CIA rats with moxibustion at the distance of 1 to 2cm, and the effect of moxibustion was better. (3) from tolerance, acceptability and curative effect. Comprehensive consideration, moxibustion 2cm group is the best distance of moxibustion for the treatment of rheumatoid arthritis rats. The results of this study provide important reference value and reference value for the clinical standardization of the application of moxibustion for the treatment of RA.
【学位授予单位】：南京中医药大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R245

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