PPAR-δ agonist affects adipo-chondrogenic differentiation of human mesenchymal stem cells through the expression of PPAR-γ,Regenerative Therapy

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PPAR-δ agonist affects adipo-chondrogenic differentiation of human mesenchymal stem cells through the expression of PPAR-γ
Regenerative Therapy ( IF 3.4 ) Pub Date : 2020-07-28 , DOI: 10.1016/j.reth.2020.07.003
Dong Hyun Kim ₁ , Dong Hwan Kim ₂ , Bruce E Heck ₃ , Michael Shaffer ₃ , Keon Hee Yoo ₁ , Jin Hur ₄

Affiliation

Introduction

Peroxisome proliferator–activated receptor (PPAR) subfamily play an important role in chondrogenesis. Previous study has reported that mixture of GW0742 (PPAR-δ agonist), hyaluronic acid (HA) and mesenchymal stem cells (MSCs) enhance chondrogenesis. The purpose of this study is to compare with efficacies of commercially available HA and demonstrate correlation of PPAR-γ and PPAR-δ.

Methods

In this experimental study, MSCs were cultured with chondrogenic media and clinical HA gels (Euflexxa®, Synvisc®, Orthovisc® and Supartz®) using micormass culture method. Expression of type Ⅰ, Ⅱ collagen and matrix metalloprotease-13 (MMP-13) was measured by immunoblotting. MSCs were cultured with chondrogenic media and/or HA and/or GW0742 and/or rosiglitazone (PPAR-γ agonist) and/or human osteoarthritis synovial fluid. Immunoblotting was used to measure expression of type Ⅱ collagen and PPAR-γ. To identify the effective dose for chondrogenesis and adipogenesis, either 0.1, 1, 5 or 10 μM of rosiglitazone was added to MSCs in chondrogenic media or adipogenic media.

Results

Clinical HA gels inhibited expression of type Ⅰ collagen and enhanced the expression of MMP-13. Type Ⅱ collagen expression was significantly elevated in all treatment groups except Supartz®. GW0742 decreased the expression of PPAR-γ with/without inflammation condition. Rosiglitazone enhanced adipogenesis in a dose-dependent manner and enhanced the expression of type Ⅱ collagen under inflammation condition. Otherwise, the expression of type Ⅱ collagen and formation of chondrocyte spheroids showed a dose-dependent manner with a peak at 1 μM of rosiglitazone.

Conclusions

PPAR-γ has a considerable anti-inflammatory effect and a strong pro-adipogenic effect, which inhibits the chondrogenic effect. PPAR-γ is related with PPAR-δ and shows a chondrogenic effect at lower concentrations. And clinical HA gels shows various efficacy of chondrogenesis. This study suggested that PPAR-γ and PPAR-δ are key regulatory factors of chondrogenesis.

中文翻译：

PPAR-δ激动剂通过表达PPAR-γ影响人间充质干细胞的脂肪软骨分化

介绍

过氧化物酶体增殖物激活受体 (PPAR) 亚家族在软骨形成中起重要作用。先前的研究报道了 GW0742（PPAR-δ 激动剂）、透明质酸 (HA) 和间充质干细胞 (MSCs) 的混合物可增强软骨形成。本研究的目的是与市售 HA 的功效进行比较，并证明 PPAR-γ 和 PPAR-δ 的相关性。

方法

在这项实验研究中，MSCs 使用微量培养法与软骨形成培养基和临床 HA 凝胶（Euflexxa®、Synvisc®、Orthovisc® 和 Supartz®）一起培养。免疫印迹法检测Ⅰ、Ⅱ型胶原和基质金属蛋白酶13（MMP-13）的表达。MSCs 与软骨形成培养基和/或 HA 和/或 GW0742 和/或罗格列酮（PPAR-γ 激动剂）和/或人骨关节炎滑液一起培养。免疫印迹法检测Ⅱ型胶原和PPAR-γ的表达。为了确定软骨形成和脂肪形成的有效剂量，将 0.1、1、5 或 10 μM 的罗格列酮添加到软骨形成培养基或脂肪形成培养基中的 MSCs。

结果

临床HA凝胶抑制Ⅰ型胶原蛋白表达，增强MMP-13表达。除Supartz®外，所有治疗组的Ⅱ型胶原蛋白表达均显着升高。GW0742 在有/无炎症条件下降低 PPAR-γ 的表达。罗格列酮以剂量依赖性方式促进脂肪生成，并在炎症条件下增强Ⅱ型胶原蛋白的表达。除此之外，Ⅱ型胶原蛋白的表达和软骨细胞球体的形成呈剂量依赖性，在罗格列酮1 μM时达到峰值。