基于TCRC的細(xì)胞毒性評(píng)估方法

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摘要 為了人們的健康與安全，日常工業(yè)化學(xué)品在進(jìn)入市場(chǎng)之前必須做深入的毒性評(píng)估試驗(yàn)。早期的毒性評(píng)估試驗(yàn)主要是活體動(dòng)物實(shí)驗(yàn)?？墒沁@種評(píng)估試驗(yàn)不僅耗時(shí)長、成本高、可靠性低，而且存在著違背動(dòng)物倫理的問題。在這種情況下，體外細(xì)胞毒性評(píng)估方法取得了很好的發(fā)展。不過，傳統(tǒng)的單點(diǎn)細(xì)胞毒性指數(shù)（如：LC50）對(duì)細(xì)胞培養(yǎng)時(shí)間依賴性太強(qiáng)。隨著細(xì)胞分析技術(shù)的進(jìn)步，人們已經(jīng)可以借助實(shí)時(shí)細(xì)胞分析儀，動(dòng)態(tài)的記錄細(xì)胞群的即時(shí)變化，從而得到隨時(shí)間變化的細(xì)胞響應(yīng)曲線。本研究首先提出使用一種雙指數(shù)模型來描述所得的細(xì)胞響應(yīng)曲線，模型中的參數(shù)k1、k2分別描述了細(xì)胞群的生長率與死亡率。然后通過回歸分析求得模型中的參數(shù)k1、k2。根據(jù)求得的結(jié)果可以發(fā)現(xiàn)參數(shù)k2與化學(xué)物質(zhì)濃度的關(guān)系，可以用S函數(shù)進(jìn)行擬合，基于此本文定義了一種新的細(xì)胞毒性指數(shù)KC50。并通過向6種細(xì)胞株中分別加入14種化學(xué)物質(zhì)的實(shí)驗(yàn)，來驗(yàn)證這種細(xì)胞毒性指數(shù)的有效性。實(shí)驗(yàn)表明：KC50細(xì)胞毒性指標(biāo)可替代諸傳統(tǒng)單點(diǎn)細(xì)胞毒性指標(biāo)，該指標(biāo)可以廣泛應(yīng)用于細(xì)胞毒性分析試驗(yàn)中，并且當(dāng)細(xì)胞毒性較低，傳統(tǒng)單點(diǎn)指標(biāo)無法評(píng)判時(shí)，該指標(biāo)仍然適用。
關(guān)鍵詞：細(xì)胞毒性，細(xì)胞響應(yīng)曲線，雙指數(shù)模型，S函數(shù)

Cytotoxicity Assessment Based on Multi-Concentration Time-Dependent Cellular Response Curves (TCRC)
Abstract In order to people's health and safety, daily chemicals must do toxicity assessment test thoroughly before entering the market.Toxicity assessment test early mainly living animal experiment. But this eva luation test is time-consuming, high cost, low reliability, and there is a violation of animal ethics.Toxicity assessment test early mainly living animal experiment. But this eva luation test is time-consuming, high cost, low reliability, and there is a violation of animal ethics.In this case, in vitro cytotoxicity eva luation method has a very good development. However, the traditional single cell toxicity index (such as: LC50) is too strong for the cell culture time dependence. With the cell analysis technology, people have already can use real-time cell analyzer, dynamic recording cell populations of instant change, so as to obtain the time-dependent cellular response curves(TCRC).This paper proposes using a two-exponent model,to describe the cell response curve, the parameters of the model are described in k1,k2 growth rate and mortality of cells. Then, through regression analysis to obtain the parameters in the model k1,k2 According to the obtained results can be found between the parameters of k2 and chemical concentration, can be fitted by sigmoidal model function, this paper defines a new cytotoxic index KC50. And through to the 6 cell lines were added to 14 kinds of chemical experiment, to verify the effectiveness of this cytotoxic index. Experimental results show: KC50 cytotoxicity index can replace the traditional single point cytotoxicity index, the index can be widely applied to the cell toxicity test, and when low cytotoxicity, the traditional single index cannot be judged, the index is still applicable.
Keywords: cytotoxicity, time-dependent cellular response curves (TCRC), two-exponent model , sigmoidal model

目 錄
第一章 緒 論 1
1.1 課題研究背景及意義 1
1.2毒性檢測(cè)發(fā)展 1
1.2.1 傳統(tǒng)動(dòng)物毒性檢測(cè)及動(dòng)物實(shí)驗(yàn)的問題 1
1.2.2 體外細(xì)胞毒性實(shí)驗(yàn) 2
1.3 細(xì)胞毒性檢測(cè)實(shí)驗(yàn)儀器 3
1.3.1 傳統(tǒng)細(xì)胞分析法所用到的實(shí)驗(yàn)儀器 3
1.3.2 實(shí)時(shí)細(xì)胞分析儀 3
1.4 幾種體外細(xì)胞毒性評(píng)估方法簡介 6
1.4.1單一時(shí)間點(diǎn)檢測(cè)方法 6
1.4.2 AUC50法 6
1.4.3 KC50法 9
1.5 本文主要研究內(nèi)容 9
1.6 本章小結(jié) 10
第二章 實(shí)驗(yàn)材料和儀器 11
2.1 細(xì)胞株 11
2.2 化學(xué)物品 11
2.3 高通量實(shí)時(shí)細(xì)胞分析法 12
2.4 本章小結(jié) 13
第三章 細(xì)胞毒性評(píng)估方法 14
3.1數(shù)據(jù)處理 14
3.2 雙指數(shù)模型 16
3.2.1模型的建立 16
3.2.2 模型的參數(shù)估計(jì) 17
3.3 KC50的方法 18
3.3.1 毒性指數(shù)k2 18
3.3.2 S函數(shù)的參數(shù)估計(jì) 19
3.4 本章小結(jié) 21
第四章 結(jié)果與討論 22
4.1 結(jié)果 22
4.2 討論 25
4.3本章小結(jié) 28
第五章 結(jié)論與展望 29
5.1結(jié)論 29
5.2展望 29
致 謝 30
參考文獻(xiàn) 31