摘要
Objective： To investigate the effects of Tounongsan （透脓散） extract （TNSE） on proliferation and apoptosis of the human lymphoma cell line Raji and its possible mechanism of action. Methods： The viability of TNSE-treated Raji cells was measured by a 3-（4,5-dimethyl-thiazol-2-yl）-2,5-diphenyltetrazolium bromide assay. Cell apoptosis was determined by flow cytometry. The molecular mechanisms of TNSE-mediated apoptosis were further investigated by reverse transcription-polymerase chain reaction （RT-PCR） analysis of the mRNA expression of nuclear factor K B （NF-κB）, Bcl-XL, Bcl-2-associated death promoter （Bad）, caspase-9 and caspase-3. Western blotting was used to detect the protein expressions of NF- κB, Bad, cleaved caspase-9 and cleaved caspase-3. Results： TNSE inhibited Raji cell proliferation in dose- and time-dependent manners. After 48-h treatment with various concentrations of TNSE （125, 250 and 500 μg/mL）, the apoptosis rates of Raji cell were 12.23% ±1.98% （P〈0.05）, 20.97% ± 3.96% （P〈0.01） and 30.4% ± 4.87% （P〈0.01）, respectively, compared with those of the control （6.02% ± 1.01%）. RT-PCR demonstrated that NF- κ B mRNA expression was significantly downregulated in Raji cells treated with 250 μg/mL TNSE for 48 h （P〈0.06）, while Bad, caspase-9 and caspase-3 mRNA levels were upregulated （P〈0.05）. Moreover, TNSE treatment resulted in downregulation of NF- κ B protein expression and strikingly upregulated protein expressions of Bad, cleaved caspase-9, cleaved caspase-3 in a dose-dependent manner, as determined by Western blot. Conclusion： TNSE exhibits significant anti-proliferative and apoptotic effects in Raji cells, which may be involved in regulation of NF- κB and Bad, and activation of caspase-9 and caspase-3.
Objective： To investigate the effects of Tounongsan （透脓散） extract （TNSE） on proliferation and apoptosis of the human lymphoma cell line Raji and its possible mechanism of action. Methods： The viability of TNSE-treated Raji cells was measured by a 3-（4,5-dimethyl-thiazol-2-yl）-2,5-diphenyltetrazolium bromide assay. Cell apoptosis was determined by flow cytometry. The molecular mechanisms of TNSE-mediated apoptosis were further investigated by reverse transcription-polymerase chain reaction （RT-PCR） analysis of the mRNA expression of nuclear factor K B （NF-κB）, Bcl-XL, Bcl-2-associated death promoter （Bad）, caspase-9 and caspase-3. Western blotting was used to detect the protein expressions of NF- κB, Bad, cleaved caspase-9 and cleaved caspase-3. Results： TNSE inhibited Raji cell proliferation in dose- and time-dependent manners. After 48-h treatment with various concentrations of TNSE （125, 250 and 500 μg/mL）, the apoptosis rates of Raji cell were 12.23% ±1.98% （P〈0.05）, 20.97% ± 3.96% （P〈0.01） and 30.4% ± 4.87% （P〈0.01）, respectively, compared with those of the control （6.02% ± 1.01%）. RT-PCR demonstrated that NF- κ B mRNA expression was significantly downregulated in Raji cells treated with 250 μg/mL TNSE for 48 h （P〈0.06）, while Bad, caspase-9 and caspase-3 mRNA levels were upregulated （P〈0.05）. Moreover, TNSE treatment resulted in downregulation of NF- κ B protein expression and strikingly upregulated protein expressions of Bad, cleaved caspase-9, cleaved caspase-3 in a dose-dependent manner, as determined by Western blot. Conclusion： TNSE exhibits significant anti-proliferative and apoptotic effects in Raji cells, which may be involved in regulation of NF- κB and Bad, and activation of caspase-9 and caspase-3.
基金
Supported by A Proiect of the Priority Academic Program Development of Jiangsu Higher Education Institutions(No. 01 2062003010)
Proiect of Top Talented Personnel in Six Profession In Jiangsu Province(No.2011-WS-049)
Project of Jiangsu Province Hospital of Traditional Chinese Medicine (No.2010-Y1008)
