本文關鍵詞：次聲對人外周血單個核細胞的影響研究　出處：《南方醫(yī)科大學》2008年碩士論文　論文類型：學位論文

  更多相關文章： 次聲 效應 人外周血單個核細胞 細胞膜

【摘要】： 背景 次聲是頻率在10~(-5)～20Hz之間的機械振動波。隨著對次聲的研究深入,次聲波在疾病診療方面的巨大開發(fā)潛力已日益受到國內外學者的重視。目前有個別學者認為一定強度的次聲能加強人的免疫系統(tǒng)功能,但鮮見關于此的實驗及機制研究。 人的免疫系統(tǒng)在抗感染和適應環(huán)境方面起重要作用,人外周血單個核細胞(peripheral blood mononuclear cell,PBMC)是免疫系統(tǒng)的重要組成部分,是機體完成細胞免疫和體液免疫的重要基礎,人外周血單個核細胞可作為反應免疫功能的一個指標,其分類技術(分離液密度梯度離心法)已十分成熟,所以人外周血單個核細胞在臨床及實驗研究得到廣泛的應用。 研究目的: 本研究擬在探討治療用次聲對健康人單個核細胞(PBMC)的生物學效應,包括對健康人PBMC的活性和細胞周期及凋亡的影響,對健康人PBMC的超微結構的影響,為次聲對人體免疫系統(tǒng)的影響提供實驗依據,為次聲治療提供相關參數。 研究假設: 本研究采用的次聲治療劑量對健康人PBMC的存在生物學效應,對健康人PBMC的細胞活性有促進作用,可改變細胞的凋亡率;次聲處理使PBMC膜超微結構的改變。 材料與方法: 每個健康志愿者為1例樣本,共20例(n=20),分離后每例分成3份:次聲組(次聲處理)、對照組(處理同次聲組但次聲治療儀處于關閉狀態(tài))、空白組(淋巴細胞分離后一直置于5%CO2飽和濕度的培養(yǎng)箱內培養(yǎng));對次聲治療儀進行消毒后,超凈臺內實驗處理15 min、30 min、60 min、90 min、120 min,每時間段依次取樣后放入5%CO2飽和濕度的培養(yǎng)箱內進行培養(yǎng)。培養(yǎng)24 h、48 h,檢測相關指標,包括:臺盼藍染色法、噻唑蘭染色法(MTT法)、流式細胞術以及掃描電鏡和透射電鏡的檢測。 實驗數據采用spss13.0進行統(tǒng)計分析,用重復測量的方差分析方法,多組間比較采用LSD方法。 結果: 1臺盼藍染色法(重復3次)結果顯示,分離后的細胞存活率達到95%以上,說明整個分離過程對PBMC的損傷不大,不會對下面進行的實驗或對照處理產生影響,可進行下一步實驗。 2采用MTT法檢測的結果顯示,實驗處理后培養(yǎng)24 h、48 h,次聲組的OD值與對照組之間的差異均無統(tǒng)計學意義(P＞0.05);但次聲組OD值隨處理時間呈依次增大趨勢,而對照組這種趨勢不明顯。此外,各組的OD值的變異系數較大。 3流式細胞儀檢測結果顯示,各次聲處理組與相應的對照組比較,凋亡細胞(早期凋亡,晚期凋亡)以及其他各類細胞的比率,其差異性均無統(tǒng)計學意義(P值均＞0.05)。 4掃描電鏡下顯示:次聲處理作用120 min/培養(yǎng)48 h在細胞的大小、形狀、表面微絨毛疏密程度與空白組差異不大;而對照組細胞卻有細胞表面微絨毛減少、異形細胞增加等變化。 5透射電鏡下顯示:次聲組與空白組相比,細胞狀態(tài)尚好,細胞核未有明顯改變,核膜清晰但不規(guī)則,線粒體略腫脹,細胞膜表面突起略減少。而對照組,細胞外形不規(guī)則,細胞微絨毛減少,細胞核呈均質化,個別細胞出現核膜模糊甚至破壞,核溶解,胞質未見結構完整清晰的細胞器,可見自噬泡,其中含有變性的胞質成分。 結論: 本研究采用的次聲治療劑量對健康人PBMC的細胞活性可能有一定促進作用,但對其凋亡率的影響不大;次聲處理可能會使細胞膜性結構產生變化;次聲處理過程可能對PBMC的自然老化過程和外界損害因素的影響有延緩作用;次聲所致的PBMC膜超微結構的改變或生物膜通透性的變化可能是其對PBMC的生物學效應的機制之一。 總結: 本實驗采用的次聲對健康人PBMC的細胞活性可能有一定促進作用,進而影響人的免疫功能。但次聲對PBMC凋亡影響不明顯,次聲對PBMC的增殖和細胞周期影響不大。次聲可導致PBMC膜超微結構的改變;次聲處理過程可能對PBMC的自然老化或凋亡過程和外界損害因素的影響有延緩作用。 本研究提示,INFRASOUND 8~(TM)次聲治療儀對健康人PBMC的生物學效應可能與次聲對細胞膜超微結構的直接作用有關,次聲可影響人的免疫系統(tǒng)功能,但次聲對免疫反應中的其他過程的影響有待進一步研究。
[Abstract]:background
Infrasound is at the frequency of 10~ (-5) - mechanical vibration wave 20Hz. Along with the study of infrasound depth, infrasound in clinical diagnosis and treatment of the huge potential for development have been paid more and more attention of scholars at home and abroad. At present, some scholars believe that the function of the immune system of a certain intensity of infrasound can enhance people's, but study on experiment and mechanism about this is rare.
The immune system plays an important role in anti infection and adapt to the environment, human peripheral blood mononuclear cells (peripheral blood mononuclear cell, PBMC) is an important part of the immune system, is an important basis for the body to complete the cellular immunity and humoral immunity, an index of human peripheral blood mononuclear cells can be used as immune function the reaction, the classification technology (separation medium density gradient centrifugation) has been very mature, so the human peripheral blood mononuclear cells have been widely used in clinical and experimental research.
The purpose of the study is:
This study was to explore the treatment of infrasound on healthy human mononuclear cells (PBMC) of biological effects, including activity and effect on cell cycle and apoptosis in healthy human PBMC, effects on ultrastructure of healthy human PBMC, to provide the experimental basis for the effect of infrasound on the human immune system, provide relevant parameters for infrasound therapy.
Research hypothesis:
The biological effect of infrasound therapy dose on healthy human PBMC is promoted by this study. It can promote the cell activity of PBMC in healthy people, and change the apoptosis rate of cells. Infrasound treatment can make the ultrastructure of PBMC membrane change.
Materials and methods:
Each healthy volunteers were 1 samples, a total of 20 cases (n=20), separated into 3 cases each: infrasound group (infrasound treatment), control group (treatment group for sound treatment instrument is closed but infrasound), control group (culture box has been placed on the 5%CO2 saturated humidity after separation of lymphocytes); infrasound therapy after disinfection, super clean bench experimental treatment for 15 min, 30 min, 60 min, 90 min, 120 min, each time in the incubator after sampling in 5%CO2 saturated humidity were cultured. After 24 h, 48 h, including the related indicators detection: trypan blue staining, thiazole blue staining method (MTT method), flow cytometry and scanning electron microscopy and transmission electron microscopy.
The experimental data were analyzed by SPSS13.0, and the method of repeated measurement was used for the analysis of variance. The LSD method was used in multiple groups.
Result:
1 trypan blue staining (repeated 3 times) showed that the survival rate of the cells after separation was over 95%, indicating that the whole separation process had little damage to PBMC and would not affect the following experiments or control treatments. Further experiments could be carried out.
2 detected by MTT showed that after the experimental treatment and cultured for 24 h, 48 h, the difference between the OD value of the infrasound group and the control group had no statistical significance (P > 0.05); but the infrasound group od with treatment time showed a trend of increasing, while the control group, this trend is not obvious. In addition, the OD value of each the coefficient of variation is larger.
3 flow cytometry results showed that there was no significant difference in the ratio of apoptotic cells (early apoptosis, late apoptosis) and other kinds of cells between the infrasound treatment group and the corresponding control group (P > 0.05).
4 scanning electron microscope showed: infrasound treatment for 120 min/ and culture for 48 h had no significant difference in cell size, shape and surface microvilli density, while in control group, there was a decrease in cell surface microvilli and an increase in heteromorphic cells.
5 transmission electron microscope: compared with infrasound group and blank control group, cell nucleus state is still good, no obvious change, but the nuclear membrane was clear and irregular, mitochondrial slightly swollen, cell membrane surface were reduced slightly. While the control group cells, irregular shape, microvilli, cell nucleus was homogenized, individual cell membrane fuzzy even destruction, nuclear dissolution, cytoplasmic organelles structure is not complete and clear, visible autophagic vacuoles, which contain cytoplasmic components of degeneration.
Conclusion:
Cell activity dose used in this study of infrasound on healthy human PBMC may have a certain role in promoting, but has little effect on the apoptosis rate; infrasound treatment may cause cell membrane structure changes; infrasound process may affect the natural aging process of PBMC and external damage factors delayed changes; or the biological membrane permeability of PBMC membrane ultrastructure caused by the infrasound could be one of the mechanisms of the biological effects of PBMC.
Summary:
In this experiment, the cell activity of infrasound on healthy human PBMC may have a certain role in promoting, thereby affecting the immune function. But the effect of infrasound on the apoptosis of PBMC is not obvious, infrasound proliferation and cell cycle of PBMC is not affected. Infrasound can cause PBMC membrane ultrastructure change; time sound processing may impact on PBMC the natural aging process and external damage or apoptosis factors have delayed effect.
This study suggested that INFRASOUND 8~ (TM) biological effect of infrasound therapy on healthy human PBMC may be related to the direct effects of infrasound on ultrastructure and cell membrane structure, infrasound can affect the human immune system function, but the effect of infrasound on other processes in the immune response remains to be further studied.

【學位授予單位】：南方醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2008
【分類號】：R363

【引證文獻】

相關碩士學位論文 前1條

1 畢迎立;次聲對骨髓間充質干細胞生長情況的影響[D];南方醫(yī)科大學;2012年

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本文編號：1375541