【摘要】：目的比較不同濃度硝普鈉(SNP)誘導(dǎo)兔軟骨細(xì)胞凋亡模型,為進(jìn)一步探討藥物對(duì)軟骨細(xì)胞凋亡的調(diào)控作用提供理論支持。方法 2014年9—12月選取4周齡健康雄性新西蘭大耳白兔8只,體質(zhì)量200~250 g,體外分離培養(yǎng)并鑒定兔軟骨細(xì)胞,采用不同濃度SNP(0、0.062 5、0.125、0.25、0.5、1、2 mmol/L)誘導(dǎo)兔軟骨細(xì)胞,分別于12、24、48 h采用四甲基偶氮唑鹽比色試驗(yàn)法(MTT法)和末端脫氧核苷酸轉(zhuǎn)移的d UTP缺口末端標(biāo)記法(TUNEL法)評(píng)價(jià)并比較不同濃度SNP誘導(dǎo)軟骨細(xì)胞凋亡模型,記錄兔軟骨細(xì)胞增殖情況(OD值)和兔軟骨細(xì)胞凋亡率。結(jié)果不同濃度SNP誘導(dǎo)的兔軟骨細(xì)胞在不同時(shí)間點(diǎn)的OD值比較,差異均有統(tǒng)計(jì)學(xué)意義(P0.05)。SNP濃度為0.062 5、0.125 mmol/L培養(yǎng)12、24、48 h時(shí)兔軟骨細(xì)胞OD值高于SNP濃度為0 mmol/L(P0.05);SNP濃度為0.25、0.5、1、2 mmol/L培養(yǎng)12、24、48 h時(shí)兔軟骨細(xì)胞OD值均低于SNP濃度為0 mmol/L;SNP濃度為0.25、0.5、1、2 mmol/L培養(yǎng)12、24、48 h時(shí)兔軟骨細(xì)胞OD值在不同濃度間兩兩比較,差異均有統(tǒng)計(jì)學(xué)意義(P0.05)。不同濃度SNP誘導(dǎo)的兔軟骨細(xì)胞在不同時(shí)間點(diǎn)的凋亡率比較,差異均有統(tǒng)計(jì)學(xué)意義(P0.05)。SNP濃度為0.25、0.5、1、2 mmol/L培養(yǎng)12、24、48 h時(shí)兔軟骨細(xì)胞凋亡率均高于SNP濃度為0 mmol/L(P0.05);且不同濃度間兩兩比較,差異有統(tǒng)計(jì)學(xué)意義(P0.05)。結(jié)論 SNP濃度為0.25~2 mmol/L且誘導(dǎo)12、24、48 h時(shí)可制備不同嚴(yán)重程度的軟骨細(xì)胞凋亡模型,為進(jìn)一步探討藥物對(duì)軟骨細(xì)胞凋亡的調(diào)控作用提供了理論支持。
[Abstract]:Objective to compare the model of rabbit chondrocyte apoptosis induced by sodium nitroprusside (SNP) at different concentrations, and to provide theoretical support for the further study of the regulation of chondrocyte apoptosis by drugs. Methods from September to December 2014, 8 healthy male New Zealand white rabbits of 4 weeks of age, weighing 200,250g, were isolated and identified as chondrocytes in vitro. The chondrocytes were isolated and identified with different concentrations of SNP. Mmol/L) induced rabbit chondrocytes, The apoptotic model of chondrocytes induced by different concentrations of SNP was evaluated and compared with tetramethylazolium colorimetric assay (MTT) and d UTP Nick end labeling (TUNEL) for terminal deoxynucleotide transfer. The proliferation of rabbit chondrocytes (OD) and the rate of chondrocyte apoptosis were recorded. Results the OD values of rabbit chondrocytes induced by different concentrations of SNP were compared at different time points. The difference was statistically significant (P0.05). SNP concentration was 0.062, 0.125 mmol/L culture for 48 h, the OD value of rabbit chondrocytes was higher than that of SNP concentration was 0 mmol/L (P0.05); The OD values of chondrocytes in rabbit chondrocytes were lower than that of SNP concentration 0 mmol/L; after cultured for 48 h with SNP concentration of 0.25 and 0.5 mmol/L. The OD value of rabbit chondrocytes was significantly higher than that of rabbit chondrocytes at different concentrations (P0.05) when the concentration of SNP was 0.25 and 0.5 / 1 / 2 mmol/L cultured for 48 h (P < 0.05) and the OD values of chondrocytes in rabbits were significantly different (P0.05). The apoptosis rate of rabbit chondrocytes induced by SNP at different concentration was significantly different at different time points (P0.05). SNP concentration was 0.25). SNP 0.51, P 0.05), and the apoptosis rate of rabbit chondrocytes induced by SNP at different time points was significantly different (P0.05). 2 the apoptotic rate of rabbit chondrocytes was higher than that of SNP (0 mmol/L) at 48 h after mmol/L culture (P0.05). And different concentrations of pairwise comparison, the difference was statistically significant (P0.05). Conclusion the model of chondrocyte apoptosis with different severity can be established when the concentration of SNP is 0.25 ~ 2 mmol/L and induced for 48 h, which provides theoretical support for further study on the regulation of chondrocyte apoptosis by drugs.
【作者單位】： 遼寧醫(yī)學(xué)院藥學(xué)院;遼寧醫(yī)學(xué)院附屬第一醫(yī)院;中國(guó)科學(xué)院上海藥物研究所;
【基金】：國(guó)家自然科學(xué)基金資助項(xiàng)目(81302681) 國(guó)家“十二五”重大新藥創(chuàng)制基金(2012ZX093016-002) 遼寧省自然科學(xué)基金資助項(xiàng)目(2013022051) 遼寧醫(yī)學(xué)院校長(zhǎng)基金-奧鴻博澤基金(XZJJ20130106-02);遼寧醫(yī)學(xué)院校長(zhǎng)基金-全民口腔研究生科研創(chuàng)新基金(QM2014007)
【分類號(hào)】：R965

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