Sickle cell disease (SCD) is a genetic blood disorder that affects the shape and transportation of red blood cells (RBCs) in blood vessels, leading to various clinical complications. Sickle cell disease is a widesprea...Sickle cell disease (SCD) is a genetic blood disorder that affects the shape and transportation of red blood cells (RBCs) in blood vessels, leading to various clinical complications. Sickle cell disease is a widespread genetic disease in Black Africa. The objective of this study was to evaluate the sickling inhibition activity of Griffonia simplicifolia. Quantitative and qualitative tests were used to determine the major groups of secondary metabolites present in the Griffonia simplicifolia leaves extracts and the modified Emmel test was used to perform the study of sickling inhibition activity. The OECD 423 toxicity study showed that at a single dose of 2000 mg/kg bw, Griffonia simplicifolia leaves extracts are not toxic. All tested substances inhibit erythrocyte falciformation in a dose-dependent manner. The percentages of inhibition were 50.35%, 73% and 94.23% for aqueous extract, hydromethanolic extract and phenylalanine respectively at the concentration of 15 mg/mL. The methanolic extract (70%) had higher activity compared to the aqueous extract. These results suggest that the Griffonia simplicifolia extracts have some potential to be used as alternative antisickling therapy in SCD management.展开更多
PCR扩增溶血素(suilysin,SLY)基因,克隆后测序,利用原核表达载体pET-28a及表达宿主菌E.coli BL21(DE3)构建SLY原核表达系统,采用SDS-PAGE检测蛋白表达情况,借助Ni-NTA亲和层析法纯化蛋白,获得具有序列保守且可溶性的SLY蛋白。显微镜形...PCR扩增溶血素(suilysin,SLY)基因,克隆后测序,利用原核表达载体pET-28a及表达宿主菌E.coli BL21(DE3)构建SLY原核表达系统,采用SDS-PAGE检测蛋白表达情况,借助Ni-NTA亲和层析法纯化蛋白,获得具有序列保守且可溶性的SLY蛋白。显微镜形态学观察显示,质量浓度为0.4 mg/L的SLY在37℃条件下作用2 h可使贴壁hCMEC/D3(人脑微血管内皮细胞)全部死亡。通过分光光度法测定SLY对人红细胞的溶血活性,结果显示1μg SLY可使2.25×10^(10)个人红细胞完全溶解。Real-time RT-PCR结果显示,SLY处理所致破碎的红细胞上清可使hCMEC/D3细胞炎性因子IL-6和IL-8转录水平分别升高80倍和100倍以上,且该作用效果显著高于超声破碎红细胞的方式。结果表明,SLY具有显著的细胞毒性,其导致红细胞溶血可能是猪链球菌2型(Streptococcus suis type 2,SS2)引起机体炎症反应的原动力。展开更多
文摘Sickle cell disease (SCD) is a genetic blood disorder that affects the shape and transportation of red blood cells (RBCs) in blood vessels, leading to various clinical complications. Sickle cell disease is a widespread genetic disease in Black Africa. The objective of this study was to evaluate the sickling inhibition activity of Griffonia simplicifolia. Quantitative and qualitative tests were used to determine the major groups of secondary metabolites present in the Griffonia simplicifolia leaves extracts and the modified Emmel test was used to perform the study of sickling inhibition activity. The OECD 423 toxicity study showed that at a single dose of 2000 mg/kg bw, Griffonia simplicifolia leaves extracts are not toxic. All tested substances inhibit erythrocyte falciformation in a dose-dependent manner. The percentages of inhibition were 50.35%, 73% and 94.23% for aqueous extract, hydromethanolic extract and phenylalanine respectively at the concentration of 15 mg/mL. The methanolic extract (70%) had higher activity compared to the aqueous extract. These results suggest that the Griffonia simplicifolia extracts have some potential to be used as alternative antisickling therapy in SCD management.
文摘PCR扩增溶血素(suilysin,SLY)基因,克隆后测序,利用原核表达载体pET-28a及表达宿主菌E.coli BL21(DE3)构建SLY原核表达系统,采用SDS-PAGE检测蛋白表达情况,借助Ni-NTA亲和层析法纯化蛋白,获得具有序列保守且可溶性的SLY蛋白。显微镜形态学观察显示,质量浓度为0.4 mg/L的SLY在37℃条件下作用2 h可使贴壁hCMEC/D3(人脑微血管内皮细胞)全部死亡。通过分光光度法测定SLY对人红细胞的溶血活性,结果显示1μg SLY可使2.25×10^(10)个人红细胞完全溶解。Real-time RT-PCR结果显示,SLY处理所致破碎的红细胞上清可使hCMEC/D3细胞炎性因子IL-6和IL-8转录水平分别升高80倍和100倍以上,且该作用效果显著高于超声破碎红细胞的方式。结果表明,SLY具有显著的细胞毒性,其导致红细胞溶血可能是猪链球菌2型(Streptococcus suis type 2,SS2)引起机体炎症反应的原动力。
