随肥胖患者增多，近年来肥胖相关性肾小球病（ORG）发病显著增加。ORG表现为蛋白尿和肾功能进行性下降，病理表现为肾小球肥大，伴或不伴节段性硬化。目前认为ORG是一种足细胞病。申请人所在团队近10年对ORG进行了一系列研究，成功建立了ORG动物及细胞模型，并进行了中、西医的干预研究。我们前期研究显示姜黄的主要成分姜黄素，能有效改善ORG足细胞损伤，减轻蛋白尿及肾小球肥大，但具体机制不详。结合文献及预实验，我们提出假说：由Wnt5a介导的Wnt/Ca2+途径激活是ORG足细胞损伤的机制之一，TRPC6可能是Wnt5a的下游效应因子；姜黄素可能通过抑制Wnt5a/TRPC6而发挥拮抗ORG足细胞的作用。本研究拟用体外培养的足细胞为对象，从分子及蛋白质水平对上述假说进行验证。本研究为原创性研究，研究成果将为临床应用姜黄素及含姜黄素中药治疗ORG提供理论基础。

Now obesity has been a global epidemic. The incidence of obesity related glomerulopathy (ORG) is also increased rapidly. ORG is now considered as a podocyte disease. The clinical manifestations of ORG are proteinuria and renal insufficiency. Renal pathological feature of ORG is the glomerular hypertrophy, with or without segmental sclerosis. Our team has focused on the study of ORG for at least twelve years. We analyzed and reported the clinicopathological features, and measured and determined normal value range of glomerular diameter in Chinese adults. We created a mouse model of ORG and performed intervention studies on this model. We have published several research papers about ORG in Chinese and international journals. Today the treatment of ORG by using western medicine is still limited. it is necessary to find some useful herb for treatment of ORG from traditional Chinese medicine. Our previous study showed that the main component of turmeric, curcumin, can antagonize the podocyte injury of ORG, but the underlie mechanism is still not clear. Based on our previous work, preliminary experiments data and literature review, we propose the hypothesis that Wnt5a mediated activation of Wnt/Ca2+ pathway is one of the mechanisms of ORG related podocyte injury, and TRPC6 may be a downstream effector of Wnt5a; curcumin may ameliorate ORG related podocyte injury through inhibiting Wnt5a/TRPC6 signal. Our study is tightly connected with our previous studies. We intend to verify the above hypothesis through podocyte experiments in vitro. This research is an original study and no such studies were reported before. If this project can be completed well, the pathogenesis of ORG should be further understanding, which could provide a novel direction for ORG treatment in the future.

随肥胖患者增多，近年来肥胖相关性肾小球病（ORG）发病显著增加。目前认为ORG是一种足细胞病。我们前期研究显示姜黄素能有效改善ORG足细胞损伤，但具体机制不详。结合文献及预实验，我们提出假说:：由Wnt5a介导的Wnt/Ca2+途径激活是ORG足细胞损伤的机制之一，TRPC6可能是Wnt5a的下游效应因子；姜黄素可能通过抑制Wnt5a/TRPC6而发挥拮抗ORG足细胞的作用。本研究通过2 个细胞实验来验证这一假说。方法：（1）将条件永生小鼠足细胞分为3 组：对照组、瘦素组、瘦素+姜黄素组。（2）将条件永生小鼠足细胞诱导分化后分为3 组：对照组、Wnt 5a刺激组、Wnt 5a+姜黄素组。应用实时荧光定量PCR 和免疫印迹法分别检测nephrin、podocin、desmin、Wnt 5a和TRPC6 的mRNA 和蛋白质表达变化。用钙离子荧光素探针Fura 2-AM检测各组细胞内钙离子浓度变化。结果：（1）与对照组相比，瘦素能抑制足细胞nephrin、podocin的mRNA和蛋白的表达，上调足细胞desmin的mRNA和蛋白的表达。加入姜黄素后，nephrin、podocin、desmin的水平恢复。瘦素能显著活化Wnt5a/TRPC6，与对照组比较，瘦素组中足细胞的Wnt5a和TRPC6的mRNA和蛋白的表达均被上调，且细胞内钙离子浓度增加。与瘦素组比较，加入姜黄素后，Wnt5a和TRPC6的mRNA和蛋白的表达均下降，且细胞内钙离子浓度下降。（2）与对照组相比，Wnt5a能抑制足细胞nephrin、podocin的mRNA和蛋白的表达，上调足细胞desmin的mRNA和蛋白的表达。加入姜黄素后，nephrin、podocin、desmin的水平恢复。Wnt5a能显著活化TRPC6，与对照组比较，Wnt5a组足细胞TRPC6的mRNA和蛋白的表达均被上调，且细胞内钙离子浓度增加。与Wnt5a组比较，加入姜黄素后，TRPC6的mRNA和蛋白的表达均下降，且细胞内钙离子浓度下降。结论：瘦素可通过Wnt5a/TRPC6途径损伤足细胞，TRPC6可能为Wnt5a的下游因子，而姜黄素可能通过抑制Wnt5a/TRPC6的活化起到保护足细胞的作用。