Gond Ling, Liu Dai-shun, Lin Jiang, Wu Yang, Zhu Hong-lan
Zhongguo Zhong Xi Yi Jie He Za Zhi. 2015 Oct;35(10):1249-54.
To explore the effect of curcumin on TGF-β2 regulated peroxisome proliferater activated receptor y (PPAR-γ)/platelet derived growth factor β (PDGF-β) signaling pathway in lung fibroblasts of mice.
C57BL/6 mouse lung fibroblasts were in vitro cultured with TGF-β2, curcumin, or TGF-β2 plus curcumin. The cell proliferation was detected by cell growth counting in the blank control group, low, middle, and high dose curcumin groups (5, 25, 50 μmol/L), the TGF-β2 (10 ng/mL) group, TGF-β2 (10 ng/mL) plus curcumin (5, 25, 50 μmol/L) groups. mRNA expressions of PPAR-γ, platelet-derived growth factor receptor β (PDGFR-β), fibroblast growth factor R1 (FGFR1) were detected using reverse transcription PCR. Protein levels of PPAR-γ and collagen-1 were detected using Western blot and ELISA in the blank control group, the TGF-β2 group, the TGF-β2 (10 ng/mL) plus curcumin 50 μmol/L group.
Compared with the blank control group, curcumin 50 μmol/L showed the most significant inhibition on cell proliferation at 48 h and 72 h. Compared with the TGF-β2 group, TGF-β2 (10 ng/mL) plus curcumin 50 mol/L also showed the most significant inhibition on cell proliferation at 48 h and 72 h. Compared with the blank control group, mRNA expressions of PPAR-γ and PDGF-β, as well as protein expression of PPAR-γ increased, the collagen-1 expression also increased in the TGF-β2 group (P < 0.05). Compared with the TGF-β2 group, mRNA expressions of PPAR-γ obviously increased in the TGF-β2 (10 ng/mL) plus curcumin 25 μmol/L group and the TGF-β2 (10 ng/mL) plus curcumin 50 μmol/L group, higher than that in the TGF-β2 (10 ng/mL) plus curcumin 5 [μmol/L group (P < 0.05). mRNA expressions of PPAR-γ was higher in the TGF-β2 (10 ng/mL) plus curcumin 50 μmol/L group than in the TGF-β2 (10 ng/mL) plus curcumin 25 μmol/L group (P < 0.05). mRNA expressions of PDGF-β was lower in TGF-β2 (10 ng/mL) plus curcumin groups than in the TGF-β2 group (P < 0.05). Besides, PDGF-β mRNA expressions were lower in the TGF-β2 (10 ng/mL) plus curcumin 50 μmol/L group than in the TGF-β2 (10 ng/mL) plus curcumin 5 μmol/L group and the TGF-β2 (10 ng/mL) plus curcumin 25 μmol/L group (P < 0.05). There was no statistical difference in FGFR1 mRNA expressions between the TGF-β2 group and 3 TGF-β2 plus curcumin groups (P > 0.05). Compared with the TGF-β2 group, PPAR-γ protein expressions increased and collagen-1 protein expressions decreased in the TGF-β2 (10 ng/mL) plus curcumin 50 μLmol/L group (P < 0.05, P < 0.01).
Curcumin not only could inhibit TGF-β2 induced proliferation of lung fibroblasts, but also could inhibit the synthesis of collagens. These might be associated with up-regulating PPAR-γ expressions and down-regulating PDGF-β expressions. Therefore, curcumin might inhibit the occurrence and developing of lung fibrosis through blocking PPAR-γ/PDGF-β signaling pathway.
探讨姜黄素对转化生长因子-β2(TGF-β2)调控的小鼠肺成纤维细胞中过氧化物酶体增殖物激活受体γ(PPAR-γ)/血小板衍生生长因子β(PDGF-β)信号通路的影响。
将C57BL/6小鼠肺成纤维细胞分别用TGF-β2、姜黄素或TGF-β2加姜黄素进行体外培养。在空白对照组、低、中、高剂量姜黄素组(5、25、50 μmol/L)、TGF-β2(10 ng/mL)组、TGF-β2(10 ng/mL)加姜黄素(5、25、50 μmol/L)组通过细胞生长计数检测细胞增殖。采用逆转录PCR检测PPAR-γ、血小板衍生生长因子受体β(PDGFR-β)、成纤维细胞生长因子R1(FGFR1)的mRNA表达。在空白对照组、TGF-β2组、TGF-β2(10 ng/mL)加50 μmol/L姜黄素组,采用蛋白质印迹法和酶联免疫吸附测定法检测PPAR-γ和胶原蛋白-1的蛋白水平。
与空白对照组相比,50 μmol/L姜黄素在48 h和72 h对细胞增殖的抑制作用最显著。与TGF-β2组相比,TGF-β2(10 ng/mL)加50 μmol/L姜黄素在48 h和72 h对细胞增殖的抑制作用也最显著。与空白对照组相比,TGF-β2组中PPAR-γ和PDGF-β的mRNA表达以及PPAR-γ的蛋白表达增加,胶原蛋白-1表达也增加(P<0.05)。与TGF-β2组相比,TGF-β2(10 ng/mL)加25 μmol/L姜黄素组和TGF-β2(10 ng/mL)加50 μmol/L姜黄素组中PPAR-γ的mRNA表达明显增加,高于TGF-β2(10 ng/mL)加5 μmol/L姜黄素组(P<0.05)。TGF-β2(10 ng/mL)加50 μmol/L姜黄素组中PPAR-γ的mRNA表达高于TGF-β2(10 ng/mL)加25 μmol/L姜黄素组(P<0.05)。TGF-β2(10 ng/mL)加姜黄素组中PDGF-β的mRNA表达低于TGF-β2组(P<0.05)。此外,TGF-β2(10 ng/mL)加50 μmol/L姜黄素组中PDGF-β的mRNA表达低于TGF-β2(10 ng/mL)加5 μmol/L姜黄素组和TGF-β2(10 ng/mL)加25 μmol/L姜黄素组(P<0.汗)。TGF-β2组与3个TGF-β2加姜黄素组之间FGFR1的mRNA表达无统计学差异(P>0.05)。与TGF-β2组相比,TGF-β2(10 ng/mL)加50 μmol/L姜黄素组中PPAR-γ蛋白表达增加,胶原蛋白-1蛋白表达降低(P<0.05,P<0.01)。
姜黄素不仅能抑制TGF-β2诱导的肺成纤维细胞增殖,还能抑制胶原蛋白的合成。这些可能与上调PPAR-γ表达和下调PDGF-β表达有关。因此,姜黄素可能通过阻断PPAR-γ/PDGF-β信号通路抑制肺纤维化的发生和发展。