He-Ne激光照射對(duì)淋巴結(jié)內(nèi)巨噬細(xì)胞、淋巴細(xì)胞影響的實(shí)驗(yàn)研究
佚名
作者;任明姬,姜麗萍,吳 巖,崔明玉
【摘要】 目的 探討弱激光對(duì)機(jī)體免疫刺激作用的機(jī)理。 用劑量63.7J/cm2的He-Ne激光照射小鼠神闕穴,照射時(shí)間為10天,每天照射5min,(1)采用臺(tái)盼藍(lán)活體注射顯示巨噬細(xì)胞吞噬力試驗(yàn)方法;(2)采用吖啶橙(AO)顯示核酸的熒光染色法分別觀察小鼠腸系膜淋巴結(jié)內(nèi)巨噬細(xì)胞的吞噬功能和淋巴細(xì)胞內(nèi)核酸含量的。(3)觀察淋巴結(jié)組織結(jié)構(gòu)。結(jié)果 (1)照射組淋巴結(jié)內(nèi)巨噬細(xì)胞吞噬功能較對(duì)照組顯著增高,差異有非常顯著性(P<0.01);(2)照射組淋巴細(xì)胞內(nèi)核酸含量較對(duì)照組明顯增多。(3)淋巴小結(jié)增多增大。結(jié)論 適當(dāng)劑量的He-Ne激光可激活淋巴結(jié)免疫功能,提高機(jī)體免疫力。 【關(guān)鍵詞】 激光;巨噬細(xì)胞; 淋巴細(xì)胞 An experimental study of He- Ne laser irradiation on the macrophages and lymphocytes in lymph node 【Abstract】 Objective To research the effects of He-Ne laser irradiation on immunity of organism.Methods Shenque acupoints of mice abdomen were irradiated by He-Ne laser at the dosages of 63.7J/cm2 for 10 days,5 minutes each day.n following methods are taken:(1)Observing the phagocytosis of the macrophages in lymph node after trypan blue injecting the live mice.(2)Observing the changes of nucleic acid of lymphocytes with fluorescence microscope.(3) Observing the tissue structure of lymph node by HE stain.Results The four indexes:number of trypan granule,area of granule,ratio of granule area to cell area and granule integral optical density at irradiated group were higher than that of non-irradiated group (P<0.01).The nucleic acid of irradiated groups were higher than non-irradiated group.In addition.the lymphoid nodule in the irradiated groups remarkable changes indicating their activation. Conclusion He-Ne laser at appropriated dosages may activate lymph node and enhance the immunity of organism. 【Key words】 laser;macrophage; lymphocyte 淋巴結(jié)是機(jī)體的主要周圍淋巴器官,位于淋巴循環(huán)的通路上,其大小、結(jié)構(gòu)及內(nèi)含細(xì)胞成分的變化與機(jī)體的免疫功能狀態(tài)密切相關(guān)。本文采用He-Ne激光照射小鼠神闕穴后,觀察激光對(duì)腸系膜淋巴結(jié)組織結(jié)構(gòu)和其內(nèi)巨噬細(xì)胞吞噬力及淋巴細(xì)胞增殖能力的影響。旨在探討低強(qiáng)度激光對(duì)機(jī)體免疫刺激作用機(jī)制,為臨床提供實(shí)驗(yàn)依據(jù)。 1 材料與方法 1.1 材料 昆明種小白鼠,體重18~22g,雌雄各半,隨機(jī)分成對(duì)照組和照射組,采用上海產(chǎn)1000B(c)He-Ne激光器照射小鼠神闕穴(剪毛),照射功率25mW,照射劑量為63.7J/cm2,光斑直徑0.3cm,照射時(shí)間為10天,每天照射5min。 1.2 方法 (1)取材前3日,每天往腹腔內(nèi)注射0.5%的無菌臺(tái)盼藍(lán)0.3ml,第4日取腸系膜淋巴結(jié)。用10%福爾馬林固定,常規(guī)法制成石蠟切片,用伊紅淡然,將切片用HI-CI型醫(yī)學(xué)圖像系統(tǒng)于每組600倍光鏡下隨機(jī)選擇12屏,統(tǒng)計(jì)巨噬細(xì)胞吞噬臺(tái)盼藍(lán)的顆粒數(shù)目(GN)、顆粒面積(GA)及其與細(xì)胞面積之比(GA/CA)和顆粒積分光密度(IOD) 4項(xiàng)指標(biāo),并輸入微機(jī)作F檢驗(yàn)。(2)取腸系膜淋巴結(jié)涂片,用Carnoy 氏固定液固定15min,再用0.01% AO緩沖液染色10min,置OLYMPUS BHF-型熒光顯微鏡的高倍鏡下觀察,DNA呈黃色熒光,RNA呈紅色熒光,DNA或RNA含量越多,黃色或紅色越深越亮。(3)常規(guī)HE染色方法觀察淋巴結(jié)的組織結(jié)構(gòu)。 2 結(jié)果 2.1 He-Ne激光對(duì)淋巴結(jié)巨噬細(xì)胞吞噬能力的影響 淋巴結(jié)的每組切片置圖像分析系統(tǒng)的光鏡下放大600倍,隨機(jī)選擇12屏觀察淋巴結(jié)髓質(zhì)區(qū),巨噬細(xì)胞胞質(zhì)內(nèi)出現(xiàn)藍(lán)色顆粒為陽性,臺(tái)盼藍(lán)顆粒呈大小、粗細(xì)不等,分布不均,顏色深淺各異。圖像分析結(jié)果用統(tǒng)計(jì)軟件作F檢驗(yàn),見表1。
表1 He-Ne激光照射對(duì)淋巴結(jié)巨噬細(xì)胞吞噬功能的圖像分析結(jié)果 (x±s,n=30)
注:照射10天組與對(duì)照組比較,*P<0.01