Pingmu Decoction Induces Orbital Preadipocytes Apoptosis In Vitro.

Yali Zhang,Hong Li,Long Gao,Xia Zhang,Ruifang Xie

doi:10.1155/2017/2109249

Abstract

Pingmu Decoction is the Traditional Chinese Medicine which has treated Graves' Ophthalmopathy (GO) in the inactive stage for more than ten years. This study was to explore the mechanism of Pingmu Decoction of inhibiting preadipocytes in GO patients from differentiating into mature adipocytes. Human orbital preadipocytes were isolated and cultured through tissue explant method. Orbital preadipocytes were induced into mature adipocytes. The medicinal serum was prepared from rats. The cells were treated with medicinal serum which were divided into three groups, low dose group (5%), medium dose group (10%), and high dose group (20%). The cells viabilities were observed by Oil Red O staining, MTT method, and Annexin V/propidium iodide (PI) double staining. Effect of Pingmu Decoction on cell apoptosis rate of orbital matured adipocytes was measured by flow cytometry. The genes Fas and Fas L from cell groups were tested by RT-PCR and Western blotting. The expression of master adipogenic transcription factors, including peroxisome proliferation-activity receptor (PPAR) γ and CCAAT/enhancer binding protein (C/EBP) α, was tested by Western blotting. Pingmu Decoction could reduce orbital preadipocytes viability and induce apoptosis of mature adipocyte via Fas/Fas L signaling pathway. Pingmu Decoction reduced lipid accumulation and downregulated the expression of PPAR γ and C/EBP α. Pingmu Decoction may play a therapeutic effect by reducing the accumulation of orbital adipocytes.

Highlights

Graves’ Ophthalmopathy (GO), known as thyroidassociated ophthalmopathy, is an autoimmune disease characterized by invasion of retrobulbar and orbital tissues [1, 2]
There were statistical differences between the medium dose group and low dose group (P < 0.05), but not the high dose group, in the early apoptosis rate. These results suggest that Pingmu Decoction serum induces apoptosis of mature GO adipocytes, with the most dramatic effect occurring at the medium dose
Our results show that the cell viability of preadipocytes could be reduced by Pingmu Decoction serum at low medium and high doses at 48 h, with the most significant effects observed with the medium dose and at 96 h

Summary

Introduction

Graves’ Ophthalmopathy (GO), known as thyroidassociated ophthalmopathy, is an autoimmune disease characterized by invasion of retrobulbar and orbital tissues [1, 2]. An increase in orbital adipose tissue content may increase the orbital pressure and lead to proptosis. Abnormal proliferation of orbital adipose tissue and the occurrence of fat in GO play an important role in disease development [7, 8]. An increase in orbital preadipocyte volume and quantity may lead to the characteristic increase in adipose tissues [9]. Recent studies have confirmed that preadipocytes that reside in orbital tissues of patients can transform into fatty cells under certain conditions and contribute to the development of GO [10, 11]. An imbalance in the proliferation, differentiation, and apoptosis of orbital preadipocytes may be the main factor regulating orbital exenteration increase, orbital pressure increase, and the occurrence of proptosis in patients with GO

Methods

Results

Conclusion