本文關(guān)鍵詞：硫酸銅和氯化鋅對(duì)不同倍性泥鰍鰭細(xì)胞系的毒性效應(yīng)　出處：《大連海洋大學(xué)》2015年碩士論文　論文類(lèi)型：學(xué)位論文

  更多相關(guān)文章： 泥鰍鰭細(xì)胞系 半致死濃度 抗氧化酶 微核率 超微結(jié)構(gòu)

【摘要】：本文研究了不同濃度的硫酸銅和氯化鋅對(duì)70-80代的二倍體泥鰍鰭細(xì)胞系(DIMF)和三倍體泥鰍鰭細(xì)胞系(TRMF)的毒性效應(yīng)。采用四噻唑藍(lán)(MTT)比色法測(cè)得DIMF和TRMF的硫酸銅的半致死濃度分別為154.3士23μmol/L、184.6士14μmol/L。對(duì)氯化鋅的半致死濃度為245.2士12μmol/L、293.6士23μmol/L。三倍體泥鰍鰭細(xì)胞系半致死濃度明顯高于二倍體泥鰍鰭細(xì)胞系(P0.05)。DIMF和TRMF中酶活性的測(cè)定結(jié)果顯示:經(jīng)硫酸銅處理后,二倍體泥鰍鰭細(xì)胞系中超氧化物歧化酶(SOD)的活性在硫酸銅濃度為0~200μmol/L范圍Qg,隨濃度的升高逐漸升高,在200μmol/L濃度時(shí)達(dá)到最大值,隨后逐漸降低。三倍體泥鰍鰭細(xì)胞系中SOD的活性,隨硫酸銅濃度的增加逐漸降低。谷胱甘肽-S-轉(zhuǎn)移酶(GST)和谷胱甘肽過(guò)氧化物酶(GSH-Px)的活性在兩種細(xì)胞系中隨著硫酸銅濃度的增加呈現(xiàn)逐漸下降的趨勢(shì)。TRMF細(xì)胞中三種酶的活性均高于DIFM。經(jīng)氯化鋅處理后,以上三種酶活性均出現(xiàn)先升高再降低的趨勢(shì),當(dāng)氯化鋅濃度為100μmol/L時(shí)酶活性達(dá)到最大值。微核試驗(yàn)檢測(cè)硫酸銅對(duì)細(xì)胞核的損傷,發(fā)現(xiàn)DIMF和TRMF細(xì)胞中均有微核產(chǎn)生,且微核率隨硫酸銅濃度增加逐漸提高。在濃度為400μmol/L時(shí)達(dá)到最大值,二倍體為3.33‰,三倍體為4.33‰。經(jīng)氯化鋅處理后,氯化鋅濃度為0~500μmol/L時(shí),微核率隨濃度增加而增加。濃度到達(dá)500μmol/L時(shí),二倍體和三倍體微核率均達(dá)到最大值,二倍體為3.33‰,三倍體是4.33‰,以后隨濃度增加沒(méi)有顯著變化(P0.1)。但在相同濃度下三倍體的微核率要高于二倍體的微核率(P0.1)。超微結(jié)構(gòu)觀察結(jié)果:對(duì)照組DIMF和TRMF中細(xì)胞器沒(méi)有明顯差異,但TRMF囊泡較多。經(jīng)過(guò)硫酸銅處理后,DIMF和TRMF細(xì)胞的病理變化情況相似,表現(xiàn)為細(xì)胞界限模糊,細(xì)胞核膜結(jié)構(gòu)不完整,線粒體嵴解體,粗面內(nèi)質(zhì)網(wǎng)膜溶解,核糖體脫落游離于胞質(zhì)中,表明細(xì)胞出現(xiàn)了壞死。經(jīng)過(guò)氯化鋅處理的細(xì)胞,二倍體與三倍體的病理變化較一致:核解體、線粒體水腫、溶酶體形成吞噬小泡、胞漿濃縮顏色較深。表現(xiàn)出細(xì)胞凋亡的特征。
[Abstract]:The toxic effects of copper sulfate and zinc chloride on 70-80 generation diploid loach fin cell line (DIMF) and triploid loach fin cell line (TRMF) were studied. The semi lethal concentration of copper sulfate with four thiazolium (MTT) colorimetric method for measuring DIMF and TRMF was 154.3, 23, and 184.6, 14, mol/L, respectively. The semi lethal concentration of zinc chloride was 245.2, 12 mol/L, 293.6 and 23 mu mol/L. The semi lethal concentration of the fin cell line of triploid loach was significantly higher than that of the diploid loach fin cell line (P0.05). The determination results of enzyme activity of DIMF and TRMF in the display: after copper sulfate treatment, diploid loach fin cells of superoxide dismutase (SOD) activity in the copper sulfate concentration was 0~200 mol/L Qg, gradually increased with the concentration increased, reached the maximum at 200 mol/L concentration, then decreased gradually. The activity of SOD in the fin cell line of triploid loach gradually decreased with the increase of copper sulfate concentration. The activity of glutathione -S- transferase (GST) and glutathione peroxidase (GSH-Px) in two cell lines showed a decreasing trend with the increase of cupric sulfate concentration. The activity of three enzymes in TRMF cells was higher than that of DIFM. After the treatment of zinc chloride, the activity of the above three enzymes increased first and then decreased. When the concentration of zinc chloride was 100 mol/L, the enzyme activity reached the maximum. Micronucleus test was used to detect the damage of copper sulfate to the nucleus. Micronuclei were found in DIMF and TRMF cells, and the micronucleus rate increased gradually with the increase of copper sulfate concentration. When the concentration is 400 mol/L, the maximum is reached, the diploid is 3.33 per 1000, and the triploid is 4.33 per thousand. After zinc chloride treatment, the micronucleus rate increases with the concentration of 0~500 micronucleus when the concentration of zinc chloride is mol/L. When the concentration reaches 500 micron mol/L, the rate of micronucleus of diploid and triploid reaches the maximum value, diploid is 3.33 per thousand, triploid is 4.33 per thousand, and it has no significant change with increasing concentration (P0.1). However, the micronucleus rate of triploid in the same concentration is higher than the micronucleus rate of diploid (P0.1). The results of ultrastructure observation: there was no significant difference in the organelles between DIMF and TRMF in the control group, but there were more TRMF vesicles. After the treatment of copper sulfate, the pathological changes of DIMF and TRMF cells were similar. The cells showed fuzzy boundaries, incomplete nuclear membrane structure, disintegration of mitochondria cristae, dissolution of rough endoplasmic reticulum, ribosome exfoliation and dissociation in cytoplasm, indicating necrosis of cells. After zinc chloride treatment, the pathological changes of diploid and triploid were consistent: nuclear disintegration, mitochondrial edema, lysosome formation, phagocytosis, and cytoplasm thickening. It shows the characteristics of cell apoptosis.
【學(xué)位授予單位】：大連海洋大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類(lèi)號(hào)】：X171.5

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