Aim: To evaluate in vitro the effectiveness of several anti-infective agents alone and in combination against Leishmania donovani. Method: A convenient stratified sampling method was used to obtain selected anti-infec...Aim: To evaluate in vitro the effectiveness of several anti-infective agents alone and in combination against Leishmania donovani. Method: A convenient stratified sampling method was used to obtain selected anti-infective agents. For individual drug samples, Half Maximal Inhibitory Concentrations (IC<sub>50</sub>) were obtained using the broth dilution method. The IC<sub>50’s</sub> of the drugs which were active against L. donovani were used as reference values to prepare drug combinations for the modified microdilution checkerboard method. Results: Five (5) out of the fifty-six (56) drugs used showed activity (inhibition of cell growth) against L. donovani cells. They include Quinine sulphate (IC<sub>50</sub> = 0.089 μg/ml), gentamicin (IC<sub>50</sub> = 8.1 μg/ml), amodiaquine (IC<sub>50</sub> = 138 μg/ml) and the two standard drugs: Amphotericin B (IC<sub>50</sub> = 6.3 μg/ml) and Pentamidine (IC<sub>50</sub> = 25 μg/ml). The remaining fifty-one (51) drugs did not show any inhibition within the range of concentrations used (1.25 - 160 μg/ml). The drug combinations of Pentamidine/Amodiaquine, Pentamidine/ Quinine sulphate, Pentamidine/Gentamicin, Amphotericin B/Quinine Sulphate, Amphotericin B/ Gentamicin, Amodiaquine/Quinine sulphate and Amodiaquine/Gentamicin showed synergistic effects against L. donovani whereas the Amphotericin B/Amodiaquine combination was antagonistic. Notable in the results obtained was the high effectiveness of quinine sulphate in inhibiting the growth of L. donovani. Quinine sulphate, though not indicated for leishmania treatment, was more effective than the two standard drugs and has a potential of playing a significant role in the treatment of leishmaniasis. Conclusion: This study has revealed five (5) anti-infective agents that by themselves or in combinations show activity against L. donovani. Some of the drug combinations which showed synergism should further be investigated. These results have to be confirmed by in vivo studies to define their roles in leishmaniasis treat展开更多
目的:探讨下调生长阻滞和DNA损伤诱导蛋白45β(growth arrest and DNA damage inducible protein 45β,GADD45β)表达对PC9肺腺癌细胞及吉非替尼敏感性的影响。方法:设计并合成GADD45β基因小干扰RNA(GADD45β-small interfering RNA,GA...目的:探讨下调生长阻滞和DNA损伤诱导蛋白45β(growth arrest and DNA damage inducible protein 45β,GADD45β)表达对PC9肺腺癌细胞及吉非替尼敏感性的影响。方法:设计并合成GADD45β基因小干扰RNA(GADD45β-small interfering RNA,GADD45β-siRNA)序列,通过慢病毒介导将GADD45β-siRNA转入PC9肺腺癌细胞中,通过实时荧光定量PCR(quantitative real-time PCR,qRT-PCR)和Western印迹检测转染前后PC9肺腺癌细胞GADD45β的mRNA及蛋白水平,采用膜联蛋白V(annexin V)-别藻蓝蛋白(allophycocyanin,APC)双染流式细胞法检测转染后细胞凋亡水平;通过流式细胞术检测转染后细胞内DNA含量,计算转染后细胞各周期时相百分率,分析转染对细胞生长周期的影响;通过计数克隆形成数检测RNA干扰对细胞成瘤能力的影响;采用MTT法检测PC9肺腺癌细胞的吉非替尼半数抑制浓度IC50。结果:筛选出5'-AAATCCACTTCACGCTCAT-3'为GADD45β基因RNA干扰的有效序列。转染GADD45β-siRNA 48 h后,qRT-PCR和Western印迹结果显示PC9肺腺癌细胞GADD45β的mRNA和蛋白表达水平明显下调(均P<0.05),细胞凋亡率明显增加(P<0.05),且成瘤克隆数明显减少(P<0.05);PC9肺腺癌细胞位于S期及G2/M期细胞增多(P<0.05),吉非替尼的IC50明显下降(P<0.05)。结论:PC9肺腺癌细胞转染GADD45β-siRNA后,能成功下调GADD45β基因的mRNA和蛋白表达;下调GADD45β表达可降低PC9肺腺癌细胞的克隆形成能力,促进细胞凋亡;下调GADD45β表达可明显提高PC9肺腺癌细胞对吉非替尼的敏感性。展开更多
文摘Aim: To evaluate in vitro the effectiveness of several anti-infective agents alone and in combination against Leishmania donovani. Method: A convenient stratified sampling method was used to obtain selected anti-infective agents. For individual drug samples, Half Maximal Inhibitory Concentrations (IC<sub>50</sub>) were obtained using the broth dilution method. The IC<sub>50’s</sub> of the drugs which were active against L. donovani were used as reference values to prepare drug combinations for the modified microdilution checkerboard method. Results: Five (5) out of the fifty-six (56) drugs used showed activity (inhibition of cell growth) against L. donovani cells. They include Quinine sulphate (IC<sub>50</sub> = 0.089 μg/ml), gentamicin (IC<sub>50</sub> = 8.1 μg/ml), amodiaquine (IC<sub>50</sub> = 138 μg/ml) and the two standard drugs: Amphotericin B (IC<sub>50</sub> = 6.3 μg/ml) and Pentamidine (IC<sub>50</sub> = 25 μg/ml). The remaining fifty-one (51) drugs did not show any inhibition within the range of concentrations used (1.25 - 160 μg/ml). The drug combinations of Pentamidine/Amodiaquine, Pentamidine/ Quinine sulphate, Pentamidine/Gentamicin, Amphotericin B/Quinine Sulphate, Amphotericin B/ Gentamicin, Amodiaquine/Quinine sulphate and Amodiaquine/Gentamicin showed synergistic effects against L. donovani whereas the Amphotericin B/Amodiaquine combination was antagonistic. Notable in the results obtained was the high effectiveness of quinine sulphate in inhibiting the growth of L. donovani. Quinine sulphate, though not indicated for leishmania treatment, was more effective than the two standard drugs and has a potential of playing a significant role in the treatment of leishmaniasis. Conclusion: This study has revealed five (5) anti-infective agents that by themselves or in combinations show activity against L. donovani. Some of the drug combinations which showed synergism should further be investigated. These results have to be confirmed by in vivo studies to define their roles in leishmaniasis treat
文摘目的:探讨下调生长阻滞和DNA损伤诱导蛋白45β(growth arrest and DNA damage inducible protein 45β,GADD45β)表达对PC9肺腺癌细胞及吉非替尼敏感性的影响。方法:设计并合成GADD45β基因小干扰RNA(GADD45β-small interfering RNA,GADD45β-siRNA)序列,通过慢病毒介导将GADD45β-siRNA转入PC9肺腺癌细胞中,通过实时荧光定量PCR(quantitative real-time PCR,qRT-PCR)和Western印迹检测转染前后PC9肺腺癌细胞GADD45β的mRNA及蛋白水平,采用膜联蛋白V(annexin V)-别藻蓝蛋白(allophycocyanin,APC)双染流式细胞法检测转染后细胞凋亡水平;通过流式细胞术检测转染后细胞内DNA含量,计算转染后细胞各周期时相百分率,分析转染对细胞生长周期的影响;通过计数克隆形成数检测RNA干扰对细胞成瘤能力的影响;采用MTT法检测PC9肺腺癌细胞的吉非替尼半数抑制浓度IC50。结果:筛选出5'-AAATCCACTTCACGCTCAT-3'为GADD45β基因RNA干扰的有效序列。转染GADD45β-siRNA 48 h后,qRT-PCR和Western印迹结果显示PC9肺腺癌细胞GADD45β的mRNA和蛋白表达水平明显下调(均P<0.05),细胞凋亡率明显增加(P<0.05),且成瘤克隆数明显减少(P<0.05);PC9肺腺癌细胞位于S期及G2/M期细胞增多(P<0.05),吉非替尼的IC50明显下降(P<0.05)。结论:PC9肺腺癌细胞转染GADD45β-siRNA后,能成功下调GADD45β基因的mRNA和蛋白表达;下调GADD45β表达可降低PC9肺腺癌细胞的克隆形成能力,促进细胞凋亡;下调GADD45β表达可明显提高PC9肺腺癌细胞对吉非替尼的敏感性。
