硼替佐米通过PPARγ途径抑制肺动脉平滑肌细胞TRPC蛋白的表达

目的验证蛋白酶体抑制剂硼替佐米(Bortezomib,BTZ)对调节肺动脉平滑肌细胞瞬时受体通道蛋白表达的影响,探讨其对调节瞬时受体通道蛋白表达的分子机制。方法原代培养大鼠肺动脉平滑肌细胞,将细胞分为常氧组、常氧+BTZ组、低氧组及低氧+BTZ组,60 h后,通过免疫印迹法测定PPARγ、TRPC1、6蛋白的表达。另将原代培养的肺动脉平滑肌细胞分为低氧组、低氧+BTZ组、低氧+BTZ+T0070907组,60 h后,通过免疫印迹法测定TRPC1、6蛋白的表达。结果(1)与常氧组相比,低氧组、低氧+BTZ组细胞TRPC1蛋白的相对表达量分别为(158±11)%和(112±8)%,低氧+BTZ组细胞TRPC1蛋白表达量相较于低氧组明显降低(P<0.05);(2)与常氧组相比,低氧组、低氧+BTZ组细胞TRPC6蛋白的相对表达量分别为(146±9)%和(107±6)%,低氧+BTZ组细胞TRPC6蛋白表达量相较于低氧组明显降低(P<0.05);(3)与常氧组相比,低氧组、低氧+BTZ组细胞PPARγ蛋白的相对表达量分别为(65±7)%和(98±6)%,低氧+BTZ组细胞PPARγ蛋白表达量相较于低氧组明显升高(P<0.05);(4)与低氧组相比,低氧+BTZ组、低氧+BTZ+T0070907组,TRPC1蛋白的相关表达量分别为(65±7)%和(92±9)%,低氧+BTZ+T0070907组细胞TRPC1蛋白表达量相较于低氧+BTZ组明显升高(P<0.05);(5)与低氧组相比,低氧+BTZ组、低氧+BTZ+T0070907组TRPC6蛋白的相关表达量分别为(71±5)%和(97±7)%,低氧+BTZ+T0070907组细胞TRPC6蛋白表达量相较于低氧+BTZ组明显升高(P<0.05)。结论硼替佐米通过抑制低氧诱导的肺动脉平滑肌细胞PPARγ的下调,调节TRPC蛋白的表达。

Bortezomib regulating TRPC expression by inhibiting PPARγdown-regulation in PASMCs

Objective To test the effect of Bortezomib(BTZ)on regulating PPARγexpression in rat PASMC,and to explore its regulating mechanism.Methods It used primary culture method for rat distal PASMCs,and they were divided into the normoxia group,the normoxia+BTZ group,the hypoxia group and the hypoxia+BTZ group.After 60 hours,the expreesion of TRPC1/6 and PPARγprotein in rat pulmonary artery smooth muscle was detected by Western blot.Another primary cultured PASMCs were divided into the hypoxia group,the hypoxia+BTZ group and the hypoxia+BTZ+T0070907 group,and the expression of TRPC1/6 was detected by Western blot after 60 hours.Results(1)Compared with the normoxia group,the level of TRPC1 protein expression was(158±11)%in the hypoxia group and(112±8)%in the hypoxia+BTZ group,which was obviously lower in the hypoxia+BTZ group than in the hypoxia group(P<0.05).(2)Compared with the normoxia group,the level of TRPC6 protein expression was(146±9)%in the hypoxia group and(107±6)%in the hypoxia+BTZ group,which was obviously lower in the hypoxia+BTZ group than in the hypoxia group(P<0.05).(3)Compared with the normoxia group,the level of PPARγprotein expression was(65±7)%in the hypoxia group and(98±6)%in the hypoxia+BTZ group,which was obviously higher in the hypoxia+BTZ group than in the hypoxia group(P<0.05).(4)Compared with the hypoxia group,the level of TRPC1 protein expression was(65±7)%in the hypoxia+BTZ group and(92±9)%in the hypoxia+BTZ+T0070907 group,which was obviously lower in the hypoxia+BTZ+T0070907 group than in the hypoxia+BTZ group(P<0.05).(5)Compared with the hypoxia group,the level of TRPC6 protein expression was(71±5)%in the hypoxia+BTZ group and(97±7)%in the hypoxia+BTZ+T0070907 group,which was obviously higher in the hypoxia+BTZ+T0070907 group than in the hypoxia+BTZ group(P<0.05).Conclusion Bortezomib can inhibit chronically hypoxic enhancement of TRPC protein expression by up-regulating PPARγin rat distal pulmonary arterial smooth muscle cells.