<br>地奥司明通过调节 SIRT3 介导的 NF-κB p65 核转位抑制炎症来改善肾纤维化,BMC Complementary Medicine and Therapies

当前位置： X-MOL 学术 › BMC Complementary Medicine and Therapies › 论文详情

Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)

地奥司明通过调节 SIRT3 介导的 NF-κB p65 核转位抑制炎症来改善肾纤维化
BMC Complementary Medicine and Therapies Pub Date : 2024-01-09 , DOI: 10.1186/s12906-023-04330-z
Wen-Man Zhao _{1,

2} , Xun-Liang Li _{1,

2} , Yuyu Zhu _{1,

2} , Rui Shi _{1,

2} , Zhi-Juan Wang _{1,

2} , Jian-Ping Xiao _{1,

2} , De-Guang Wang _{1,

2}

Affiliation

背景

肾纤维化被认为是一种不可逆的病理过程，是各类慢性肾脏病和肾功能衰竭发展的最终共同途径。地奥司明是一种天然黄酮苷，具有抗氧化、抗炎和抗纤维化活性。然而，地奥司明是否通过抑制肾纤维化来保护肾脏尚不清楚。我们的目的是研究地奥司明在肾间质纤维化中的作用并探讨其潜在机制。

方法

建立UUO小鼠模型，地奥司明（50 mg/kg·d和100 mg/kg·d）灌胃14 d。采用HE染色、Masson染色、免疫组织化学、蛋白质印迹和PCR来评估肾组织损伤和纤维化。采用Elisa试剂盒检测肾组织中IL-1β、IL-6、TNF-α的表达水平以及SIRT3的活性。此外，RNA 测序的富集图谱分析了信号通路的变化。在体外，用 TGF-β1 刺激人肾小管上皮细胞 (HK-2)，然后用地奥司明 (75 μM) 处理。检测细胞中SIRT3蛋白和mRNA的表达水平。此外，3-TYP（SIRT3的选择性抑制剂）和SIRT3小干扰RNA（siRNA）被用来降低HK-2中的SIRT3水平。

结果

地奥司明可减轻 UUO 诱导的肾纤维化和 TGF-β1 诱导的 HK-2 纤维化。此外，地奥司明还降低肾组织和 HK-2 培养基上清液中 IL-1β、IL-6 和 TNF-α 的水平。有趣的是，地奥司明给药增加了 UUO 肾脏中 SIRT3 的酶活性。此外，地奥司明在体外和体内均显着增加 SIRT3 的 mRNA 和蛋白表达。使用 3-TYP 或 SIRT3 siRNA 抑制 SIRT3 表达，消除了地奥司明在 HK-2 细胞中的抗炎作用。 RNA测序富集图谱分析表明，地奥司明干预组的核因子-κB（NF-κB）信号通路受到抑制。此外，我们发现TGF-β1增加了核NF-κB p65的核表达，但对NF-κB p65的总细胞内表达影响不大。此外，地奥司明还能减少 TGF-β1 引起的 NF-κB p65 核易位。 SIRT3 siRNA 敲低 SIRT3 表达增加了 NF-κB p65 的核表达，并消除了地奥司明对 NF-κB p65 表达的抑制作用。

结论

地奥司明可通过激活 SIRT3 抑制 NF-κB P65 的核转位来减少肾脏炎症和纤维化。

"点击查看英文标题和摘要"

Diosmin ameliorates renal fibrosis through inhibition of inflammation by regulating SIRT3-mediated NF-κB p65 nuclear translocation

Background

Renal fibrosis is considered an irreversible pathological process and the ultimate common pathway for the development of all types of chronic kidney diseases and renal failure. Diosmin is a natural flavonoid glycoside that has antioxidant, anti-inflammatory, and antifibrotic activities. However, whether Diosmin protects kidneys by inhibiting renal fibrosis is unknown. We aimed to investigate the role of Diosmin in renal interstitial fibrosis and to explore the underlying mechanisms.

Methods

The UUO mouse model was established and gavaged with Diosmin (50 mg/kg·d and 100 mg/kg·d) for 14 days. HE staining, Masson staining, immunohistochemistry, western blotting and PCR were used to assess renal tissue injury and fibrosis. Elisa kits were used to detect the expression levels of IL-1β, IL-6, and TNF-α and the activity of SIRT3 in renal tissues. In addition, enrichment maps of RNA sequencing analyzed changes in signaling pathways. In vitro, human renal tubular epithelial cells (HK-2) were stimulated with TGF-β1 and then treated with diosmin (75 μM). The protein and mRNA expression levels of SIRT3 were detected in the cells. In addition, 3-TYP (selective inhibitor of SIRT3) and SIRT3 small interfering RNA (siRNA) were used to reduce SIRT3 levels in HK-2.

Results

Diosmin attenuated UUO-induced renal fibrosis and TGF-β1-induced HK-2 fibrosis. In addition, Diosmin reduced IL-1β, IL-6, and TNF-α levels in kidney tissues and supernatants of HK-2 medium. Interestingly, Diosmin administration increased the enzymatic activity of SIRT3 in UUO kidneys. In addition, Diosmin significantly increased mRNA and protein expression of SIRT3 in vitro and in vivo. Inhibition of SIRT3 expression using 3-TYP or SIRT3 siRNA abolished the anti-inflammatory effects of diosmin in HK-2 cells. Enrichment map analysis by RNA sequencing indicates that the nuclear factor-kappa B (NF-κB) signaling pathway was inhibited in the Diosmin intervention group. Furthermore, we found that TGF-β1 increased the nuclear expression of nuclear NF-κB p65 but had little significant effect on the total intracellular expression of NF-κB p65. Additionally, Diosmin reduced TGF-β1-caused NF-κB p65 nuclear translocation. Knockdown of SIRT3 expression by SIRT3 siRNA increased the nuclear expression of NF-κB p65 and abolished the inhibition effect of Diosmin in NF-κB p65 expression.