目的研究蛇葡萄素(ampelopsin,AMP)诱导人宫颈癌细胞SiHa发生自噬及其可能的作用机制。方法单丹磺酰胺(monodansylcadaverine,MDC)染色法观察AMP对SiHa和C-33A细胞自噬的影响;透射电子显微镜(transmission electron microscope,TEM)观察...目的研究蛇葡萄素(ampelopsin,AMP)诱导人宫颈癌细胞SiHa发生自噬及其可能的作用机制。方法单丹磺酰胺(monodansylcadaverine,MDC)染色法观察AMP对SiHa和C-33A细胞自噬的影响;透射电子显微镜(transmission electron microscope,TEM)观察经AMP干预的SiHa和C-33A细胞超微结构的变化;自噬双标腺病毒转染(mRFP-GFP-LC3)法检测SiHa细胞自噬流强度;Western blot法对SiHa细胞中自噬相关蛋白LC3Ⅱ/Ⅰ、P62、Beclin-1、Atg13及PI3K/AKT/mTOR信号通路蛋白的表达情况进行检测。结果MDC染色结果显示,随着AMP浓度的升高,代表自噬小体的亮绿色逐渐增多;TEM实验结果表明,与空白对照组相比,AMP组细胞线粒体嵴溶解消失,同时可见自噬小体和自噬溶酶体;mRFP-GFP-LC3实验结果提示,随着AMP浓度的升高,细胞内自噬小体和自噬溶酶体增多,提示自噬流增强;同时发现LC3Ⅱ/Ⅰ、Beclin-1、Atg13蛋白表达上调,P62、p-PI3K、p-AKT、p-mTOR蛋白表达下调,表明AMP可能通过抑制PI3K/AKT/mTOR信号通路诱导SiHa细胞发生自噬;加入3-MA后,与AMP组相比,3-MA和AMP联合组的p-PI3K、p-AKT、p-mTOR的表达水平上升,提示自噬被抑制后,PI3K/AKT/mTOR信号通路被逆转。结论AMP可诱导SiHa细胞发生自噬,其机制可能与抑制PI3K/AKT/mTOR信号通路的激活有关。展开更多
Meplazumab,a humanized CD147 antibody,has shown favourable safety and efficacy in our previous clinical studies.In DEFLECT(NCT04586153),167 patients with severe COVID-19 were enroled and randomized to receive three do...Meplazumab,a humanized CD147 antibody,has shown favourable safety and efficacy in our previous clinical studies.In DEFLECT(NCT04586153),167 patients with severe COVID-19 were enroled and randomized to receive three dosages of meplazumab and a placebo.Meplazumab at 0.12 mg/kg,compared to the placebo group,showed clinical benefits in significantly reducing mortality by 83.6%(2.4%vs.14.6%,p=0.0150),increasing the proportion of patients alive and discharged without supplemental oxygen(82.9%vs.70.7%,p=0.0337)and increasing the proportion of patients who achieved sustained clinical improvement(41.5%vs.31.7%).The response rate in the 0.2 mg/kg group was relatively increased by 16.0%compared with the placebo group(53.7%vs.46.3%).Meplazumab also reduced the viral loads and multiple cytokine levels.Compare with the placebo group,the 0.3 mg/kg significantly increased the virus negative rate by 40.6%(p=0.0363)and reduced IL-8 level(p=0.0460);the 0.2 mg/kg increased the negative conversion rate by 36.9%,and reduced IL-4(p=0.0365)and IL-8 levels(p=0.0484).In this study,the adverse events occurred at a comparable rate across the four groups,with no unexpected safety findings observed.In conclusion,meplazumab promoted COVID-19 convalescence and reduced mortality,viral load,and cytokine levels in severe COVID-19 population with good safety profile.展开更多
文摘目的研究蛇葡萄素(ampelopsin,AMP)诱导人宫颈癌细胞SiHa发生自噬及其可能的作用机制。方法单丹磺酰胺(monodansylcadaverine,MDC)染色法观察AMP对SiHa和C-33A细胞自噬的影响;透射电子显微镜(transmission electron microscope,TEM)观察经AMP干预的SiHa和C-33A细胞超微结构的变化;自噬双标腺病毒转染(mRFP-GFP-LC3)法检测SiHa细胞自噬流强度;Western blot法对SiHa细胞中自噬相关蛋白LC3Ⅱ/Ⅰ、P62、Beclin-1、Atg13及PI3K/AKT/mTOR信号通路蛋白的表达情况进行检测。结果MDC染色结果显示,随着AMP浓度的升高,代表自噬小体的亮绿色逐渐增多;TEM实验结果表明,与空白对照组相比,AMP组细胞线粒体嵴溶解消失,同时可见自噬小体和自噬溶酶体;mRFP-GFP-LC3实验结果提示,随着AMP浓度的升高,细胞内自噬小体和自噬溶酶体增多,提示自噬流增强;同时发现LC3Ⅱ/Ⅰ、Beclin-1、Atg13蛋白表达上调,P62、p-PI3K、p-AKT、p-mTOR蛋白表达下调,表明AMP可能通过抑制PI3K/AKT/mTOR信号通路诱导SiHa细胞发生自噬;加入3-MA后,与AMP组相比,3-MA和AMP联合组的p-PI3K、p-AKT、p-mTOR的表达水平上升,提示自噬被抑制后,PI3K/AKT/mTOR信号通路被逆转。结论AMP可诱导SiHa细胞发生自噬,其机制可能与抑制PI3K/AKT/mTOR信号通路的激活有关。
基金The DEFLECT is supported by grants from National Natural Science Foundation of China(No.92169211).Jiangsu Pacific Meinuoke Biopharmaceuticals provided meplazumab.The views expressed in this article is the authors’opinion,not the opinion or policy of funder.
文摘Meplazumab,a humanized CD147 antibody,has shown favourable safety and efficacy in our previous clinical studies.In DEFLECT(NCT04586153),167 patients with severe COVID-19 were enroled and randomized to receive three dosages of meplazumab and a placebo.Meplazumab at 0.12 mg/kg,compared to the placebo group,showed clinical benefits in significantly reducing mortality by 83.6%(2.4%vs.14.6%,p=0.0150),increasing the proportion of patients alive and discharged without supplemental oxygen(82.9%vs.70.7%,p=0.0337)and increasing the proportion of patients who achieved sustained clinical improvement(41.5%vs.31.7%).The response rate in the 0.2 mg/kg group was relatively increased by 16.0%compared with the placebo group(53.7%vs.46.3%).Meplazumab also reduced the viral loads and multiple cytokine levels.Compare with the placebo group,the 0.3 mg/kg significantly increased the virus negative rate by 40.6%(p=0.0363)and reduced IL-8 level(p=0.0460);the 0.2 mg/kg increased the negative conversion rate by 36.9%,and reduced IL-4(p=0.0365)and IL-8 levels(p=0.0484).In this study,the adverse events occurred at a comparable rate across the four groups,with no unexpected safety findings observed.In conclusion,meplazumab promoted COVID-19 convalescence and reduced mortality,viral load,and cytokine levels in severe COVID-19 population with good safety profile.
