为了降低天然大分子功能蛋白的分离纯化成本,以琼脂为原料自制弱阴离子交换层析介质DEAE-琼脂凝胶微球。将DEAE-琼脂凝胶微球应用于坛紫菜R-藻红蛋白（R-PE）和螺旋藻C-藻蓝蛋白（C-PC）的提取分离,最佳洗脱条件为：层析柱规格h=35.0 cm...为了降低天然大分子功能蛋白的分离纯化成本,以琼脂为原料自制弱阴离子交换层析介质DEAE-琼脂凝胶微球。将DEAE-琼脂凝胶微球应用于坛紫菜R-藻红蛋白（R-PE）和螺旋藻C-藻蓝蛋白（C-PC）的提取分离,最佳洗脱条件为：层析柱规格h=35.0 cm,φ=1.0 cm,V=27.5 m L;流速v=0.33 m L/min;洗脱缓冲液为0.01 mol/L磷酸盐溶液（含有0.05~0.5 mol/L Na Cl溶液）;0.01 mol/L磷酸盐缓冲液（含0.1 mol/L Na Cl）洗脱出纯度为5.0的坛紫菜R-PE;0.01 mol/L磷酸盐缓冲液（含0.2 mol/L Na Cl）洗脱出纯度为4.4的螺旋藻C-PC。自制层析介质成本低廉、耗能少、可大量制备、分离效果好,可用于坛紫菜R-PE、螺旋藻C-PC的产业化制备。展开更多
Polycystic ovary syndrome(PCOS),a common female endocrine disorder marked by disrupted folliculogenesis and hyperandrogenism,is increasingly linked to oxidative stress.Despite this association,the mechanistic basis re...Polycystic ovary syndrome(PCOS),a common female endocrine disorder marked by disrupted folliculogenesis and hyperandrogenism,is increasingly linked to oxidative stress.Despite this association,the mechanistic basis remains poorly characterized.C-Phycocyanin(C-PC),a cyanobacteria-derived protein with potent antioxidant properties,has demonstrated therapeutic potential for treating PCOS,though the molecular pathways mediating its effects have yet to be delineated.This study employed both a dehydroepiandrosterone(DHEA)-induced murine model and DHEA-challenged human granulosa cells(KGN) to elucidate the regulatory role of C-PC.In vivo,oral administration of C-PC restored estrous cyclicity,reduced the prevalence of cystic follicles,and normalized circulating levels of testosterone,estradiol,progesterone,and luteinizing hormone(LH).In vitro,C-PC treatment activated the NRF2/x CT/GPX4 pathway,enhanced antioxidant activity,improved mitochondrial function,and suppressed ferroptotic death.Direct molecular interaction between C-PC and NRF2 was validated through molecular docking and cellular thermal shift assays(CETSA).Correspondingly,in vivo administration alleviated oxidative stress,inhibited ferroptosis,and increased GPX4 and x CT expression,effects reversed by pharmacological inhibition(ML385) and genetic silencing(AAV-sh-NRF2) of NRF2.C-PC also reduced DHEA-induced phosphorylation of AMPK,while co-treatment with an AMPK activator attenuated its effects on GPX4 and x CT,abolishing its anti-ferroptotic protection against granulosa cells.These findings suggest that C-PC mitigates PCOS pathology by repressing granulosa cell ferroptosis through coordinated activation of NRF2 and modulation of redox-sensitive AMPK signaling,highlighting its potential as a redoxtargeted therapeutic strategy for PCOS.展开更多
文摘为了降低天然大分子功能蛋白的分离纯化成本,以琼脂为原料自制弱阴离子交换层析介质DEAE-琼脂凝胶微球。将DEAE-琼脂凝胶微球应用于坛紫菜R-藻红蛋白（R-PE）和螺旋藻C-藻蓝蛋白（C-PC）的提取分离,最佳洗脱条件为：层析柱规格h=35.0 cm,φ=1.0 cm,V=27.5 m L;流速v=0.33 m L/min;洗脱缓冲液为0.01 mol/L磷酸盐溶液（含有0.05~0.5 mol/L Na Cl溶液）;0.01 mol/L磷酸盐缓冲液（含0.1 mol/L Na Cl）洗脱出纯度为5.0的坛紫菜R-PE;0.01 mol/L磷酸盐缓冲液（含0.2 mol/L Na Cl）洗脱出纯度为4.4的螺旋藻C-PC。自制层析介质成本低廉、耗能少、可大量制备、分离效果好,可用于坛紫菜R-PE、螺旋藻C-PC的产业化制备。
基金supported by the National Natural Science Foundation of China (82501986)Anhui Provincial Natural Science Foundation Youth Project (2508085QE176)+1 种基金Anhui Key Project Fund for College and University (2024AH050713)Henan Provincial Department of Science and Technology (252300421643)。
文摘Polycystic ovary syndrome(PCOS),a common female endocrine disorder marked by disrupted folliculogenesis and hyperandrogenism,is increasingly linked to oxidative stress.Despite this association,the mechanistic basis remains poorly characterized.C-Phycocyanin(C-PC),a cyanobacteria-derived protein with potent antioxidant properties,has demonstrated therapeutic potential for treating PCOS,though the molecular pathways mediating its effects have yet to be delineated.This study employed both a dehydroepiandrosterone(DHEA)-induced murine model and DHEA-challenged human granulosa cells(KGN) to elucidate the regulatory role of C-PC.In vivo,oral administration of C-PC restored estrous cyclicity,reduced the prevalence of cystic follicles,and normalized circulating levels of testosterone,estradiol,progesterone,and luteinizing hormone(LH).In vitro,C-PC treatment activated the NRF2/x CT/GPX4 pathway,enhanced antioxidant activity,improved mitochondrial function,and suppressed ferroptotic death.Direct molecular interaction between C-PC and NRF2 was validated through molecular docking and cellular thermal shift assays(CETSA).Correspondingly,in vivo administration alleviated oxidative stress,inhibited ferroptosis,and increased GPX4 and x CT expression,effects reversed by pharmacological inhibition(ML385) and genetic silencing(AAV-sh-NRF2) of NRF2.C-PC also reduced DHEA-induced phosphorylation of AMPK,while co-treatment with an AMPK activator attenuated its effects on GPX4 and x CT,abolishing its anti-ferroptotic protection against granulosa cells.These findings suggest that C-PC mitigates PCOS pathology by repressing granulosa cell ferroptosis through coordinated activation of NRF2 and modulation of redox-sensitive AMPK signaling,highlighting its potential as a redoxtargeted therapeutic strategy for PCOS.
