【摘要】：目的： 研究香菇多糖對輻射所致小鼠免疫應(yīng)激損傷的保護作用。從整體水平驗證香菇多糖的防護干預(yù)作用,并從細胞及分子水平探討香菇多糖該作用的機制。 方法： 1.實驗?zāi)Ｐ徒⑴c分組：①輻射損傷模型的建立：輻射條件：全身性輻射,吸收劑量為40cGy/d,吸收劑量率為300cGy/min,皮源距為150cm,照射野為20cm×20cm,連續(xù)照射30d。②實驗分組：實驗小鼠分為五組,分別為：輻射模型組、正常對照組、香菇多糖高劑量組、香菇多糖中劑量組和香菇多糖低劑量組。 2.T淋巴細胞提�。簯�(yīng)用尼龍毛柱法從小鼠脾臟提取T淋巴細胞。 3.NO含量測定：按照試劑盒說明書要求進行操作,測定不同組別小鼠T淋巴細胞分泌NO含量。 4.IL-2含量測定：按照劑盒說明書要求進行操作,測定不同組別小鼠T淋巴細胞分泌IL-2含量。 5.RNA提取：按照試劑盒說明書要求,從小鼠T淋巴細胞中提取mRNA. 6.RNA逆轉(zhuǎn)錄合成cDNA:按照試劑盒說明書要求,從提取出的RNA合成cDNA。 7.cDNA擴增：按照試劑盒說明說要求,采用熒光染料嵌合法,配制PCR反應(yīng)液,進行PCR擴增。 結(jié)果： 在整體水平,與正常組相比,慢性低劑量輻射可導(dǎo)致小鼠體重增加速率下降,脾臟指數(shù)和胸腺指數(shù)顯著下降,脾臟出現(xiàn)明顯的病理性改變。與輻射組相比,香菇多糖組可明顯改善上述改變。在細胞水平,與正常組相比,慢性低劑量輻射可導(dǎo)致小鼠T淋巴細胞活性降低,IL-2分泌減少,NO含量升高。與輻射組比較,香菇多糖組T淋巴細胞活性明顯增加,IL-2分泌增加,NO含量降低,且與正常組比較,無明顯差異。在分子水平,與正常組相比,慢性低劑量輻射可引起K+通道Kv1.3、Ca2+通道Orai1基因表達出現(xiàn)明顯上調(diào)。與正常組相比,香菇多糖低劑量組和高劑量組Kv1.3、Orai1基因表達無明顯差異。與輻射模型組相比,香菇多糖低劑量組Kv1.3基因表達下調(diào)、Orai1基因表達無顯著差異,香菇多糖高劑量組Kv1.3、Orail基因表達均明顯下調(diào)。 結(jié)論： 慢性低劑量輻射可導(dǎo)致小鼠整體及免疫系統(tǒng)功能改變,影響T淋巴細胞功能,引起T淋巴細胞膜上部分離子通道m(xù)RNA的表達改變,而香菇多糖可明顯干預(yù)輻射所致的上述改變。表明,香菇多糖作為一種免疫調(diào)節(jié)劑,對輻射所致小鼠免疫系統(tǒng)T淋巴細胞損傷具有明顯防護作用,而該作用可能與香菇多糖影響T淋巴細胞膜上的離子通道有一定關(guān)系。
[Abstract]:Objective: to study the protective effect of lentinan on immune stress injury induced by radiation in mice. The protective effect of Lentinus edodes polysaccharides (Lentinus edodes) was verified from the whole level, and the mechanism of Lentinus edodes polysaccharides was discussed from cell and molecular level. Methods: 1. The establishment of experimental model and group 1 radiation injury model: radiation condition: systemic radiation, absorbed dose was 40cGy / d, absorbed dose rate was 300cGy / min, skin source distance was 150cm, irradiation field was 20cm 脳 20cm, 30d.2 experiment group: experiment mice were divided into five groups. Respectively: radiation model group, normal control group, lentinan high dose group, Lentinus edodes polysaccharide medium dose group and lentinan low dose group. 2. T lymphocyte extraction: extraction of T lymphocytes from mouse spleen by nylon hair column method. Determination of 3.NO content. The content of NO secreted by T lymphocytes in different groups of mice was determined. The content of 4.IL-2 was determined. The content of IL-2 secreted by T lymphocytes from different groups of mice was determined. 5.RNA extraction: according to the requirements of kit instructions, mRNA. 6.RNA reverse transcription synthesis cDNA: was extracted from T lymphocytes of mice. Synthetic cDNA. 7.cDNA amplification from extracted RNA: according to the requirements of the kit, the PCR reaction solution was prepared by fluorescence dye embedding method, and the PCR amplification was carried out. Results: compared with the normal group, chronic low dose radiation could decrease the weight gain rate, spleen index and thymus index, and induce pathological changes in the spleen. Compared with radiation group, lentinan group could significantly improve the above changes. At the cell level, chronic low dose radiation could reduce the activity of T lymphocytes and decrease the secretion of IL-2 and increase the content of no. Compared with the radiation group, the activity of T lymphocytes in the lentinan group was significantly increased and IL-2 secretion was increased, the no content was decreased, and there was no significant difference between the Lentinus edodes group and the normal group. At the molecular level, chronic low dose radiation induced a significant up-regulation of K channel Kv1.3,Ca2 channel Orai1 gene expression. There was no significant difference in Kv1.3,Orai1 gene expression between low dose and high dose groups of Lentinus edodes polysaccharides compared with normal group. Compared with the radiation model group, there was no significant difference in the down-regulation of Kv1.3 gene expression in the low dose group of Lentinus edodes polysaccharides, but the down-regulation of Kv1.3,Orail gene expression in the high dose group of Lentinus edodes polysaccharide. Conclusion: chronic low dose radiation can change the whole and immune system function of mice, affect the function of T lymphocytes and change the expression of mRNA in some ion channels on T lymphocyte membrane. Lentinus edodes polysaccharides can significantly interfere with the radiation-induced changes. The results showed that Lentinus edodes polysaccharide, as an immunomodulator, had an obvious protective effect on T lymphocyte damage induced by radiation in mice, which might be related to the effect of lentinan on the ion channels on T lymphocyte membrane.
【學位授予單位】：青島大學
【學位級別】：碩士
【學位授予年份】：2013
【分類號】：R818.74;R-332;R965

【參考文獻】

相關(guān)期刊論文 前10條

1 劉曉麗;蔡東聯(lián);贊登;袁靜玨;鄭慧;;酵母多糖對S_(180)荷瘤小鼠抗氧化作用和免疫機能的影響[J];氨基酸和生物資源;2009年04期

2 王鵬云;王賽貞;林樹錢;林志彬;;靈芝孢子和破壁孢子多糖對體外培養(yǎng)的小鼠脾淋巴細胞及腹腔巨噬細胞免疫調(diào)節(jié)活性的比較[J];北京大學學報(醫(yī)學版);2005年06期

3 彭煦;范曉靜;石賢愛;陳劍鋒;郭養(yǎng)浩;孟春;;香菇多糖對小鼠脾T/B淋巴細胞TLR4、TLR9表達的影響[J];中國臨床藥理學雜志;2008年03期

4 遲淑萍;白冰科;戚揚;由莉越;高蓉;汪辰;程云;李進;;內(nèi)毒素對人巨噬細胞系U937細胞增殖和腫瘤壞死因子-α分泌影響的實驗研究[J];解放軍藥學學報;2010年04期

5 柳迎華;李明春;;香菇多糖注射液對慢性低劑量輻射損傷小鼠的防護作用[J];解放軍藥學學報;2012年03期

6 萬志紅,陳惠孫,陸松敏;內(nèi)毒素血癥小鼠巨噬細胞膜脂的改變及其對炎癥介質(zhì)產(chǎn)生的影響[J];中華急診醫(yī)學雜志;2003年04期

7 姚金鳳,王志新,張曉勇,張瑞峰;黃芪多糖對小鼠腹腔巨噬細胞免疫功能的調(diào)節(jié)作用研究[J];河南大學學報(醫(yī)學版);2005年01期

8 來偉;褚忠華;;Kv通道對巨噬細胞功能影響的研究進展[J];嶺南現(xiàn)代臨床外科;2008年02期

9 羅琦;梁慶模;;天然藥物抗輻射損傷作用機制的研究現(xiàn)狀與進展[J];遼寧中醫(yī)藥大學學報;2011年03期

10 翟瑞仁;逄朝霞;;熱療與低劑量輻射對小鼠免疫相關(guān)指標的影響[J];實用醫(yī)藥雜志;2011年11期

，

本文編號：2246849