聲動(dòng)力治療巨噬細(xì)胞通過(guò)促進(jìn)atp釋放趨化單核細(xì)胞科研訓(xùn)練論文(已修改)
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【正文】 哈爾濱醫(yī)科大學(xué) 七年制學(xué)生基礎(chǔ)階段科研論文 病理生理學(xué)教研室單位代碼：10226 學(xué)號(hào)：2009172057分類號(hào)： 2009172033 2009172075哈爾濱醫(yī)科大學(xué)七年制學(xué)生基礎(chǔ)階段科研訓(xùn)練論文題 目 ： 聲動(dòng)力治療巨噬細(xì)胞通過(guò)促進(jìn)ATP釋放趨化單核細(xì)胞 學(xué)科、專業(yè) ： 病理生理學(xué)教研室 學(xué) 生 姓 名： 曹博然 張翰 趙雪竹 指 導(dǎo) 教 師： 田野 教授 指 導(dǎo) 研 究 生： 孫鑫 郭淑媛 陳海波 二○一二年七月目 錄目 錄 1中文摘要 2Abstract 3文獻(xiàn)綜述 5前 言 12材料與方法 13技術(shù)路線 15結(jié) 果 16討 論 19結(jié) 論 21致 謝 21參考文獻(xiàn) 22 中文摘要目 的：證明聲動(dòng)力治療(sonodynamic therapy,SDT)巨噬細(xì)胞能夠趨化單核細(xì)胞的聚集并探討其機(jī)制。方 法：將實(shí)驗(yàn)對(duì)象分為對(duì)照組、SDT組、ATPase組及SDTATPase組。其中對(duì)照組為單純THP1源性巨噬細(xì)胞，而SDT組是在對(duì)照組基礎(chǔ)上加入終濃度為15μg/ml的5氨基酮戊酸（5Aminolevulinic acid, ALA），孵育2小時(shí)后接受聲動(dòng)力治療。ATPase組及SDTATPase組則是分別在對(duì)照組和SDT組基礎(chǔ)上加入ATPase。將實(shí)驗(yàn)對(duì)象離心后取其細(xì)胞上清液并分別用transwell方法研究其對(duì)單核細(xì)胞的趨化作用；將實(shí)驗(yàn)對(duì)象分為對(duì)照組、SDT組、zVADfmk組及SDTzVAD組。其中對(duì)照組為單純THP1源性巨噬細(xì)胞，SDT組是在對(duì)照組基礎(chǔ)上加入 ALA，孵育2小時(shí)后接受聲動(dòng)力治療。zVADfmk組及SDTzVAD組則是分別在對(duì)照組和SDT組基礎(chǔ)上加入N苯甲基氧化碳酰纈氨酸丙氨酸天冬氨酸氯化丙酮(zVADfmk)。將實(shí)驗(yàn)對(duì)象離心后，分別用熒光素酶方法檢測(cè)細(xì)胞上清及細(xì)胞內(nèi)ATP量的變化。結(jié) 果：用transwell方法研究聲動(dòng)力治療后的細(xì)胞上清對(duì)單核細(xì)胞的趨化作用時(shí)，從募集的單核細(xì)胞數(shù)量來(lái)看，聲動(dòng)力治療組明顯高于對(duì)照組（p），加入ATPase并進(jìn)行聲動(dòng)力治療組與聲動(dòng)力治療組相比顯著降低，對(duì)照組與ATPase組無(wú)統(tǒng)計(jì)學(xué)差異（p）；用熒光素酶方法，檢測(cè)聲動(dòng)力治療巨噬細(xì)胞后及加入凋亡抑制劑后細(xì)胞上清及細(xì)胞內(nèi)ATP量的變化得出：聲動(dòng)力治療組明顯高于對(duì)照組，約為其3倍（p）；加入zVADfmk并進(jìn)行聲動(dòng)力治療組較聲動(dòng)力組顯著降低；對(duì)照組與zVADfmk無(wú)統(tǒng)計(jì)學(xué)差異（p）。 結(jié) 論：聲動(dòng)力治療巨噬細(xì)胞通過(guò)促進(jìn)凋亡相關(guān)的ATP釋放趨化單核細(xì)胞 關(guān)鍵詞：ATP 動(dòng)脈粥樣硬化 聲動(dòng)力療法 巨噬細(xì)胞 AbstractPurpose: This paper is designed to investigate the effects of monocytes recruitment after sonodynamic therapy and delve into its mechanism.Methods: The subjects were divided into four groups: The control group,the SDT group, the ATPase group and the SDTATPase control group merely contains THP1 derived SDT one,containing macrophages and sonosensitizer ALA ,was incubated with fresh medium for 6 h before being exposed to pulse ultrasound irradiation. The ATPase group was a mixture of the control group and ATPase. Similarly,the SDTATPase group , was prised of the SDT group and ATPase. Cellfree supernatants from each group were assessed for their ability to attract THP1 monocytes in a transwell migration a further study of ATP concentrations changes, we use another four groups of subjects: The control group(macrophages alone),the SDT group, the zVADfmk group(added caspaseinhibitor zAVDfmk ) and the SDTzAVD group（like the SDT group pretreated with zAVDfmk）.Supernatants and cell debris after high speed centrifugation from these four groups were investigated respectively.Results: Cellfree supernatants with SDT were assessed for their ability to attract THP1 monocytes in a transwell migration assay. The SDT group induced an approximately sixfold greater recruitment of monocytes than the control group did(p), while the monocytes migration of the SDTATPase group decreased significantly pared to the SDT , there was no statistic difference between the control group and the ATPase group(p). Luminometer was used to measure the change of ATP ATP concentration of the SDT group was increased about therefold in parison with the control group (p),and the SDTzAVD group had a distinct decrease pared with the SDT group. The control group and the zVADfmk group showed no statistic difference(p).Conclusions: This study demonstrates that SDT could induce the recruitment of monocytes by promoting the release of
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