【作者】 黄瑶；

【导师】 张勤修；

【作者基本信息】 成都中医药大学 ， 中医五官科学， 2020， 硕士

【摘要】 目的:本研究从迎香穴穴位埋线对大鼠鼻腔黏膜上皮细胞的树突状细胞与AR的发病机制出发,建立AR大鼠模型,分组进行迎香穴穴位埋线、假埋线以及丙酸氟替卡松干预,干预后进行行为学评分测定,并提取所有大鼠的鼻黏膜和股动脉血清。检测鼻黏膜组织的病理情况,Western blot分析CD80、CD86的表达情况,通过ELISA检测血清中s Ig E、IL-2、IL-4、IL-5、IL-6、IL-10和IL-12细胞因子的分泌水平,探讨迎香穴穴位埋线通过作用于DC而调节Th1/Th2的平衡,调节鼻腔黏膜的高反应状态,从而达到缓解和治疗AR目的。方法:采购40只健康的SD大鼠,随机分为A组空白组、B组AR模型组、C组丙酸氟替卡松组、D组穴位埋线组、E组假埋线组,每组8只。除空白组以外的所有大鼠均用卵清蛋白联合氢氧化铝造模法制作AR大鼠模型。模型成功制作后,AR模型组和空白组不予干预;丙酸氟替卡松组以丙酸氟替卡松滴鼻,每日1次,共2周;穴位埋线组迎香穴埋线,每周1次,连续2周;假埋线组行埋线假动作,不埋线,每周1次,连续2周。观察并记录各组大鼠相关行为学进行打分,通过ELISA检测血清中s Ig E、IL-2、IL-4、IL-5、IL-6、IL-10和IL-12细胞因子的分泌水平,HE染色检测鼻黏膜组织的病理情况,Western blot分析各组大鼠鼻黏膜组织CD80、CD86蛋白表达情况,最后对所有结果进行统计分析。结果:1.各组大鼠AR相关行为学评分结果:与A组相比,B组、E组行为学评分评分明显升高,具有统计学意义(P<0.05);和B组、E组相比,C组和D组评分都明显降低,差异具有统计学意义(P<0.05),并且组间无明显差异(P>0.05)。2.光镜下各组大鼠鼻黏膜组织HE染色显示:B组、E组鼻黏膜纤毛排列杂乱,倒伏、甚至脱落,出现较多淋巴细胞和嗜酸性粒细胞,黏膜下腺体增生明显。与B组相比,C组和E组炎症反应较轻。3.ELISA检测血清s Ig E结果显示:A组与C组差异不显著(P>0.05),但是分别与B、D和E组都有显著差异(P<0.05);B组与E组差异不显著(P>0.05);C组与D组差异不显著(P>0.05)。4.ELISA检测血清中IL-2、IL-4、IL-5、IL-6、IL-10和IL-12细胞因子水平:A组与C组、B组与E组、C组与D组、A组与D组(不包括IL-10)都是两组之间差异不显著(P>0.05),A组分别与B组和E组存在差异(P<0.05),IL-10中A组与D组存在差异(P<0.05)。5.Western blot分析大鼠鼻黏膜组织CD80、CD86蛋白表达情况:A组空白对照组与C组丙酸氟替卡松治疗组以及D组穴位埋线组聚类为同一子集,差异不显著(P>0.05);B组AR模型组与E组假埋线组聚类为同一子集,差异不显著(P>0.05)。其余各组之间差异显著。结论:1.迎香穴穴位埋线能明显改善AR大鼠相关症状,降低血清s Ig E,减轻鼻黏膜的炎性反应,且和丙酸氟替卡松治疗相比无明显统计学差异(P>0.05)。2.迎香穴穴位埋线可以降低AR大鼠的CD80、CD86蛋白的表达水平,降低s Ig E和IL-4、IL-5、IL-6水平,提高IL-2、IL-10和IL-12的水平,使机体失衡的Th1/Th2状态得到纠正,最终缓解AR的临床症状,达到治疗的目的。更多还原

【Abstract】 Purpose:From this study Yingxiang point acupuncture point bury line of dendritic cells of rat nasal mucosa epithelial cells and the pathogenesis of AR,AR rat model was established,the grouping of Yingxiang point acupuncture point bury line,fake embedded wire and fluticasone propionate intervention,ethology score determined after intervention,and extract all the rat nasal mucosa and femoral artery blood serum.Nasal mucosa tissue pathological situation,detected by western blot detection the expression of CD80 and CD86,by ELISA detection in serum s Ig E,、IL-2、IL-4、IL-5、IL-6、IL-10 and the level of IL-12 the secretion of cytokines,explore Yingxiang point acupuncture point bury line by acting on DC and regulating Th1/Th2 balance,adjust the high reaction condition of nasal mucosa,so as to achieve mitigation and treatment of AR.Method:40 healthy SD rats were randomly divided into blank group A,AR model group B,flutekassone propionate group C,acupoint line embedding group D,and false line embedding group E,with 8 rats in each group.All the rats except the blank group were modeled with ovalbumin combined with aluminum hydroxide.After the model was successfully made,the AR model group and blank group did not intervene.Fluticasone propionate group received nasal drops of fluticasone propionate once a day for 2 weeks.Line embedding in Yingxiang point of acupoint embedding group was performed once a week for consecutive 2 weeks.False embedding line group line embedding feint,no embedding line,once a week,consecutive 2 weeks.Observe and record each related behavior in rats,by ELISA detection in serum s Ig E,、IL-2、IL-4、IL-5、IL-6、IL-10 and the level of IL-12 the secretion of cytokines,HE staining detection of nasal mucosa tissue pathology,Western blot analysis between groups in the rat nasal mucosa tissue CD80,CD86 protein expression,finally,all the results were statistically analyzed.Result:1.Results of AR related behavioral scores of rats in each group: compared with group A,the behavioral scores of group B and group E were significantly increased,which was statistically significant(P < 0.05).Compared with group B and group E,the scores of group C and group D were significantly lower(P < 0.05),and is no significant difference between the groups(P > 0.05).2.Under the light microscope,HE staining of the nasal mucosa tissues of rats in each group showed that: in group B and group E,the nasal mucosa cilia were disordered,prostrate and even exfoliated,with more lymphocytes and eosinophils,and submucosal gland hyperplasia was obvious.Compared with group B,group C and group E had less inflammation.3.The results of serum s Ig E detected by ELISA showed that the difference between group A and group C is not significant(P>0.05),but they are significantly different from group B,D,and E(P<0.05);group B and group E are not significantly different(P>0.05),there was no significant difference between group C and group D(P> 0.05).4.ELESA detection in serum IL-2、IL-4、IL-5、IL-6、IL-10 and IL-12 cytokines level: group A and group C,group B and group E,group C and D,group A and group D(not including IL-10)is no significant difference between two groups(P>0.05),group A and group B and group E difference(P<0.05),IL-10 differences between group A and group D(P<0.05).5.Western blot analysis of the expression of CD80 and CD86 proteins in rat nasal mucosal tissues: The cluster of the blank control group of group A was the same subset as the fluticasone propionate treatment group of group C and the acupoint embedding line group of group D,with no significant difference(P>0.05).The clustering of AR model group B and false embedded line group E was the same subset,and the difference was not significant(P>0.05).The differences between the remaining groups were significant.Conclusion:1.Acupoint catgut embedding at Yingxiang could significantly improve the symptoms of AR rats,reduce serum s Ig E and reduce the inflammatory response of nasal mucosa,and there was no significant statistical difference compared with the treatment of fluticasone propionate(P>0.05).2.Embedding wires at Yingxiang acupoint can reduce the expression levels of CD80 and CD86 proteins in AR rats,reduce the levels of s Ig E and IL-4,IL-5 and IL-6,and increase the levels of IL-2、IL-10 and IL-12,so as to correct the imbalance of Th1/Th2 in the body,eventually relieve the clinical symptoms of AR and achieve the purpose of treatment.更多还原