目的 运用网络药理学探索治疗骨关节炎(osteoarthritis,OA)的潜在药物并进行实验验证。方法 通过数据库及文献验证获取OA疾病靶点,根据候选靶点使用中药系统药理学数据库与分析平台(Traditional Chinese Medicine Systems Pharmacology ...目的 运用网络药理学探索治疗骨关节炎(osteoarthritis,OA)的潜在药物并进行实验验证。方法 通过数据库及文献验证获取OA疾病靶点,根据候选靶点使用中药系统药理学数据库与分析平台(Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform,TCMSP)挖掘作用于以上靶点的化合物及包含化合物的中药,通过构建可视化网络,筛选核心靶点及化合物进行分子对接。根据分子对接结果选择化合物进行体外实验,选取大鼠关节软骨细胞,通过CellTiter-Glo(CTG)检测各化合物对软骨细胞活力影响,确定后续实验给药浓度。使用白细胞介素-1β(IL-1β)诱导软骨细胞建立OA细胞模型,将细胞分为对照组、OA组及给药组,使用流式细胞术检测化合物抗OA软骨细胞凋亡能力,并选择其中抗凋亡效果最显著的化合物,使用蛋白印迹法检测其对软骨细胞凋亡相关蛋白MDM2、P53、Bax表达水平的影响。结果 通过数据库检索、筛选及文献验证得到87个OA靶点,其中15个靶点匹配到符合药代动力学参数(ADME)和Lipinski筛选标准的化合物,并得到746个候选化合物,从数据库中筛选得到包含以上化合物的中药433种。通过靶点、化合物、中药可视化网络获得核心靶点前列腺素G/H合成酶2(PTGS2)、雌激素受体(ESR1)、肿瘤坏死因子(TNF)、白细胞介素-6(IL6)、IL-1β,核心化合物28个。以上核心靶点和核心化合物分子对接结果显示,C-3位为羟基的甾体化合物与核心靶点表现出良好的对接效果,根据分子对接结果从候选化合物中选择3种C-3位为羟基的甾体化合物菜油甾醇、豆甾醇、薯蓣皂苷元进行体外实验验证其治疗OA的效果。体外实验发现,与OA组比较,菜油甾醇组、豆甾醇组和薯蓣皂苷元组OA软骨细胞凋亡率显著下降(P<0.05),其中菜油甾醇抑制效果最佳。蛋白印迹法检测发现,与对照组比较,OA组MDM2蛋白表达降低,P53、Bax蛋白�展开更多
BACKGROUND Crumbs homolog 2(CRB2)is a recently discovered gene that is closely related to the maintenance of normal polarity in podocytes;mutations can directly lead to steroid-resistant nephrotic syndrome(SRNS).Howev...BACKGROUND Crumbs homolog 2(CRB2)is a recently discovered gene that is closely related to the maintenance of normal polarity in podocytes;mutations can directly lead to steroid-resistant nephrotic syndrome(SRNS).However,the characteristics of nephrotic syndrome(NS)caused by CRB2 mutations have not been described.CASE SUMMARY We report a novel compound heterozygous mutation of the CRB2 gene in two siblings with SRNS.The two siblings had edema,proteinuria,hypoproteinemia and hyperlipidemia.Both their father and mother had normal phenotypes(no history of NS).Whole exon sequencing(WES)of the family showed a novel compound heterozygous mutation,c.2290(exon 8)C>T and c.3613(exon 12)G>A.Glucocorticoid therapy(methylprednisolone pulse therapy or oral prednisone)and immunosuppressive agents(tacrolimus)had no effect.During a 3-year follow-up after genetic diagnosis by WES,proteinuria persisted,but the patient was healthy.CONCLUSION CRB2 mutations related to SRNS often occur in exons 7,10,and 12.Clinical manifestations of SRNS caused by CRB2 mutations are often less severe than in other forms of SRNS.展开更多
文摘目的 运用网络药理学探索治疗骨关节炎(osteoarthritis,OA)的潜在药物并进行实验验证。方法 通过数据库及文献验证获取OA疾病靶点,根据候选靶点使用中药系统药理学数据库与分析平台(Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform,TCMSP)挖掘作用于以上靶点的化合物及包含化合物的中药,通过构建可视化网络,筛选核心靶点及化合物进行分子对接。根据分子对接结果选择化合物进行体外实验,选取大鼠关节软骨细胞,通过CellTiter-Glo(CTG)检测各化合物对软骨细胞活力影响,确定后续实验给药浓度。使用白细胞介素-1β(IL-1β)诱导软骨细胞建立OA细胞模型,将细胞分为对照组、OA组及给药组,使用流式细胞术检测化合物抗OA软骨细胞凋亡能力,并选择其中抗凋亡效果最显著的化合物,使用蛋白印迹法检测其对软骨细胞凋亡相关蛋白MDM2、P53、Bax表达水平的影响。结果 通过数据库检索、筛选及文献验证得到87个OA靶点,其中15个靶点匹配到符合药代动力学参数(ADME)和Lipinski筛选标准的化合物,并得到746个候选化合物,从数据库中筛选得到包含以上化合物的中药433种。通过靶点、化合物、中药可视化网络获得核心靶点前列腺素G/H合成酶2(PTGS2)、雌激素受体(ESR1)、肿瘤坏死因子(TNF)、白细胞介素-6(IL6)、IL-1β,核心化合物28个。以上核心靶点和核心化合物分子对接结果显示,C-3位为羟基的甾体化合物与核心靶点表现出良好的对接效果,根据分子对接结果从候选化合物中选择3种C-3位为羟基的甾体化合物菜油甾醇、豆甾醇、薯蓣皂苷元进行体外实验验证其治疗OA的效果。体外实验发现,与OA组比较,菜油甾醇组、豆甾醇组和薯蓣皂苷元组OA软骨细胞凋亡率显著下降(P<0.05),其中菜油甾醇抑制效果最佳。蛋白印迹法检测发现,与对照组比较,OA组MDM2蛋白表达降低,P53、Bax蛋白�
文摘BACKGROUND Crumbs homolog 2(CRB2)is a recently discovered gene that is closely related to the maintenance of normal polarity in podocytes;mutations can directly lead to steroid-resistant nephrotic syndrome(SRNS).However,the characteristics of nephrotic syndrome(NS)caused by CRB2 mutations have not been described.CASE SUMMARY We report a novel compound heterozygous mutation of the CRB2 gene in two siblings with SRNS.The two siblings had edema,proteinuria,hypoproteinemia and hyperlipidemia.Both their father and mother had normal phenotypes(no history of NS).Whole exon sequencing(WES)of the family showed a novel compound heterozygous mutation,c.2290(exon 8)C>T and c.3613(exon 12)G>A.Glucocorticoid therapy(methylprednisolone pulse therapy or oral prednisone)and immunosuppressive agents(tacrolimus)had no effect.During a 3-year follow-up after genetic diagnosis by WES,proteinuria persisted,but the patient was healthy.CONCLUSION CRB2 mutations related to SRNS often occur in exons 7,10,and 12.Clinical manifestations of SRNS caused by CRB2 mutations are often less severe than in other forms of SRNS.
