本文選題：電針 + 足三里　； 參考：《湖北中醫(yī)藥大學(xué)》2017年碩士論文

【摘要】：目的越來越多的研究表明,電針療法有抗過敏和抗炎作用,但對(duì)于遲發(fā)性超敏反應(yīng)(DTH)的作用機(jī)制不明。本研究旨在通過觀察電針對(duì)小鼠DTH治療效果,試探討電針足三里防治Th1介導(dǎo)的DTH的效應(yīng)機(jī)制。方法使用雌性C57BL/6小鼠,共32只,體重約為18-20g,將小鼠通過編號(hào)查詢隨機(jī)數(shù)字表隨機(jī)分為對(duì)照組8只、模型組(OVA-DTH組)8只、電針治療組(DTH+EA組)8只、假電針組(DTH+Sham組)8只共4組。測(cè)量小鼠足墊厚度,使用蘇木精和伊紅(HE)染色觀察足墊炎性細(xì)胞浸潤(rùn)情況;酶聯(lián)免疫吸附測(cè)定法(ELISA)檢測(cè)血清OVA特異性抗體IgG和IgE水平,并檢測(cè)足墊組織勻漿上清炎癥細(xì)胞因子(IFN-γ,TNF-α,IL-4和IL-5)水平;流式細(xì)胞儀檢測(cè)各組脾臟CD4~+IFN-γ+T細(xì)胞和CD4~+IL-4+T細(xì)胞數(shù);蛋白質(zhì)印跡法檢測(cè)脾臟分離純化的T細(xì)胞T-bet與GATA-3蛋白表達(dá),并比較各組間的差異。結(jié)果通過組織形態(tài)學(xué)觀察,與模型組相比,電針治療組能明顯減小OVA導(dǎo)致的小鼠足墊厚度(P0.05),假電針組與模型組相比無明顯差異(P0.05);電針治療組小鼠足墊組織真皮層炎性細(xì)胞浸潤(rùn)與聚集水平明顯低于模型組(P0.01),假電針組與模型組相比無明顯差異(P0.05);與模型組比較,電針治療組可顯著減輕OVA導(dǎo)致的小鼠足墊表皮層腫脹(P0.05)。通過酶聯(lián)免疫吸附測(cè)定法(ELISA)檢測(cè),與模型組相比,電針治療組可顯著抑制血清OVA特異性抗體IgG水平和IgE水平(P0.01),假電針組與模型組相比無明顯差異(P0.05);與模型組比較,電針治療組小鼠足墊組織Th1類細(xì)胞因子水平(如IFN-γ和TNF-α)顯著降低(IFN-γ:P0.05,TNF-α:P0.01),假電針組與模型組相比無明顯差異(P0.05);對(duì)比模型組,電針治療組和假電針組小鼠足墊組織Th2類細(xì)胞因子(IL-4,IL-5)水平無顯著差異(P0.05)。通過流式細(xì)胞學(xué)的檢測(cè),與對(duì)照組比較,模型組中脾CD4~+IFN-γ+T細(xì)胞明顯增加(P0.05),同時(shí)與對(duì)照組、模型組相比,電針治療組脾CD4~+IFN-γ+T細(xì)胞數(shù)量明顯降低,差異具有統(tǒng)計(jì)學(xué)意義(P0.01),假電針組與模型組相比無明顯差異(P0.05);而對(duì)比對(duì)照組、模型組和電針治療組三組間脾CD4~+IL-4+T細(xì)胞計(jì)數(shù)水平無明顯差異(P0.05);依據(jù)脾CD4~+IFN-γ+T細(xì)胞和脾CD4~+IL-4+T細(xì)胞計(jì)數(shù)的變化水平,與對(duì)照組、模型組相比,電針治療組的DTH小鼠脾Th1/Th2細(xì)胞比率(即脾CD4~+IFN-γ+T細(xì)胞數(shù)/脾CD4~+IL-4+T細(xì)胞數(shù))降至正常值附近,且與模型組相比,差異具有統(tǒng)計(jì)學(xué)意義(P0.05)。通過蛋白質(zhì)印跡法(Westen blot)檢測(cè),與對(duì)照組比較,模型組T-bet蛋白活性增強(qiáng),而電針治療組T-bet蛋白活性減弱,對(duì)GATA-3則無顯著影響(P0.05);通過T-bet/GATA-3比率的統(tǒng)計(jì)分析,電針治療組與模型組相比,電針治療組比率明顯低于模型組(P0.05),假電針組與模型組無明顯差異(P0.05)。結(jié)論1.電針“足三里”對(duì)OVA誘導(dǎo)的小鼠DTH炎癥具有明顯的抑制作用。2.電針“足三里”可通過減少IFN-γ的轉(zhuǎn)錄因子T-bet蛋白的表達(dá),重建Th1/Th2平衡,改善Th1介導(dǎo)的過敏性皮膚炎癥。3.電針“足三里”可作為過敏性炎癥和Th1介導(dǎo)的炎癥反應(yīng)的有效療法。
[Abstract]:Objective more and more studies have shown that electroacupuncture therapy has anti allergic and anti inflammatory effects, but the mechanism of delayed hypersensitivity (DTH) is unknown. The purpose of this study was to observe the effect of Electroacupuncture on DTH in mice and to explore the effect mechanism of Zusanli on the prevention and treatment of Th1 mediated DTH by electroacupuncture. Methods 32 female C57BL/6 mice were used. About 18-20g, the mice were randomly divided into 8 control groups, 8 rats in model group (group OVA-DTH), 8 in group DTH+EA, 8 in group DTH+Sham (group DTH+Sham) in 4 groups. The foot pad thickness of mice was measured and the infiltration of foot pad inflammatory cells was observed with hematoxylin and eosin (HE) staining; enzyme linked immunosorbent assay ( ELISA) the levels of serum OVA specific antibody IgG and IgE were detected, and the levels of inflammatory cytokines (IFN-, TNF- a, IL-4 and IL-5) in the foot pad tissue homogenate were detected. The number of CD4~+IFN- gamma +T cells and CD4~+IL-4+T cells in each group was detected by flow cytometry; the expression of T cells and proteins in the splenic isolation and purification was detected by Western blot. Compared with the model group, the electroacupuncture group could significantly reduce the foot pad thickness (P0.05) caused by OVA in the electroacupuncture group compared with the model group, and there was no significant difference between the sham acupuncture group and the model group (P0.05), and the level of inflammatory cell infiltration and aggregation in the foot pad tissue of the electroacupuncture group was significantly lower than that of the model group (P0.01). There was no significant difference between the model group and the model group (P0.05). Compared with the model group, the electroacupuncture treatment group could significantly reduce the mouse foot pad epidermal swelling (P0.05) caused by OVA. The electroacupuncture treatment group could significantly inhibit the serum OVA specific antibody IgG level and IgE level (P0.01) by enzyme linked immunosorbent assay (ELISA), compared with the model group. There was no significant difference between the model group and the model group (P0.05). Compared with the model group, the level of Th1 cell factors (such as IFN- gamma and TNF- alpha) in the foot pad tissue of the electroacupuncture group decreased significantly (IFN-: P0.05, TNF- A: P0.01), and there was no significant difference (P0.05) in the sham acupuncture group compared with the model group (P0.05); the contrast model group, the electroacupuncture treatment group and the sham acupuncture group mice feet. There was no significant difference in the level of Th2 cell factor (IL-4, IL-5) in the padding tissue (P0.05). Compared with the control group, the CD4~+IFN- gamma +T cells in the model group were significantly increased by flow cytometry (P0.05). Compared with the control group, the number of IFN- gamma +T cells in the spleen CD4~+ IFN- +T cells in the electroacupuncture group was significantly lower than that in the model group. The difference was statistically significant (P0.01). There was no significant difference between the pseudo electroacupuncture group and the model group (P0.05), but there was no significant difference in the CD4~+IL-4+T cell count level between the three groups in the control group and the electroacupuncture group (P0.05), and the level of the CD4~+IFN- gamma +T cell and the spleen CD4~+IL-4+T cell count was compared with the control group and the model group, the DTH mouse spleen Th1/ in the electroacupuncture group. The ratio of Th2 cells (the number of spleen CD4~+IFN- +T cells / splenic CD4~+IL-4+T cells) dropped to the normal value, and compared with the model group, the difference was statistically significant (P0.05). Compared with the control group, the viability of T-bet protein in the model group was enhanced compared with the control group, but the activity of T-bet protein in the electroacupuncture group was weakened, and GATA-3 was less than that of the control group. There was no significant effect (P0.05). Through the statistical analysis of the T-bet/GATA-3 ratio, the ratio of electroacupuncture treatment group was significantly lower than that of model group (P0.05), and there was no significant difference between the electroacupuncture group and the model group (P0.05). Conclusion 1. electroacupuncture "Zusanli" had a significant inhibitory effect on the DTH inflammation induced by OVA in the OVA induced mice, ".2. acupuncture", "foot three" By reducing the expression of the IFN- gamma transcription factor T-bet protein, reconstructing the Th1/Th2 balance and improving the Th1 mediated allergic skin inflammation,.3. electroacupuncture "Zusanli" can be used as an effective therapy for allergic inflammation and Th1 mediated inflammatory reaction.

【學(xué)位授予單位】：湖北中醫(yī)藥大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R245.97

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