Therapeutic Effects of Conditioned Medium of Neural Differentiated Human Bone Marrow-Derived Stem Cells on Rotenone-Induced Alpha-Synuclein Aggregation and Apoptosis.

Mahesh Ramalingam,Han-Seong Jeong,Sujeong Jang,Yohei Hayashi

doi:10.1155/2021/6658271

Mahesh Ramalingam, Han-Seong Jeong + Show 2 more

Open Access

https://doi.org/10.1155/2021/6658271

Copy DOI

Abstract

Mesenchymal stem cells (MSCs) have been used against several diseases. Their potential mainly appears from its secreted biomolecules. Human bone marrow-derived stem cells (hBMSC) displayed neuronal functional characteristics after differentiation by basic fibroblast growth factor (bFGF) and forskolin. PD is a chronic age-related neurodegenerative disease (NDD) characterized by loss of dopaminergic neurons in the substantia nigra (SN) and abnormal accumulation of α-synuclein (α-syn) aggregations. In this present study, we evaluated the therapeutic effects of neural differentiated hBMSC (NI-hBMSC) conditioned medium (NI-hBMSC-CM) to a rotenone- (ROT-) induced Parkinson's disease (PD) model in SH-SY5Y cells. NI-hBMSC-CM treatment (50% diluted) in the last 24 h of 48 h ROT (0.5 μM) toxicity showed a significant increase in cell survival. The decreased tyrosine hydroxylase (TH) expression as a hallmark of PD was increased by NI-hBMSC-CM. The Triton X-100-soluble and Triton X-100-insoluble cell lysate fractions were used in Western blotting. The oligomeric, dimeric, and monomeric phosphorylated serine129 (p-S129) α-syn and total monomeric α-syn were decreased during ROT toxicity in the Triton X-100-soluble fraction. The Triton X-100-insoluble fraction revealed that ROT toxicity significantly increased the oligomeric but decreased the dimeric and monomeric p-S129 α-syn expressions while all forms of total α-syn were increased in SH-SY5Y cells. NI-hBMSC-CM stabilized the physiological α-syn monomers and reduced aggregated insoluble p-S129 α-syn against ROT. The cytoskeletal proteins, neurofilament-H (NF-H), β3-tubulin (Tuj1), neuronal nuclei (NeuN), and synaptophysin (SYP) were significantly decreased during ROT toxicity. In addition, proapoptotic Bax was increased by ROT with decreased antiapoptotic Bcl-2 and Mcl-1 as well as proforms of caspase-9, caspase-3, caspase-7, and PARP-1. NI-hBMSC-CM ameliorated the neurotrophic protein expressions, controlled the Bax/Bcl-2 ratio, upregulated procaspases, and inactivated PARP-1. From our results, we conclude that NI-hBMSC-CM containing released biomolecules during neural differentiation employs regenerative effects on the ROT model of PD in SH-SY5Y cells.

Highlights

Mesenchymal stem cells (MSCs) can be found and isolated from many different body tissues, including bone marrow, placenta, adipose tissue [1], umbilical cord, bone trabeculae, muscle, synovium, dental pulp, and periodontal ligament [2]
SH-SY5Y cells were exposed with or without ROT (0.5 μM) for 24 h; the culture medium was removed; a new culture medium with or without Human bone marrow-derived stem cells (hBMSC)-conditioned medium (CM) or NI-hBMSCCM at 100, 50, and 25% was diluted in Dulbecco’s modified Eagle’s medium (DMEM) +1% fetal bovine serum (FBS) incubated in the absence or presence of ROT (0.5 μM) for another 24 h (Figure 1(a))
NI-hBMSC-CM treatment with 100, 50, and 25% dilution against ROT toxicity showed significantly increased cell numbers; on the contrary, treatment of hBMSC-CM did not show any significant changes against ROT toxicity

Summary

Introduction

Mesenchymal stem cells (MSCs) can be found and isolated from many different body tissues, including bone marrow, placenta, adipose tissue [1], umbilical cord, bone trabeculae, muscle, synovium, dental pulp, and periodontal ligament [2]. MSC are attractive therapeutic candidates in the treatment of several diseases including neurodegenerative diseases (NDD) [3, 5]. Evidence confirms that neuroprotection of MSC appears from its secretion of different proteins, including growth factors, cytokines, chemokines, Stem Cells International metabolites, and bioactive lipids, which have paracrine and autocrine therapeutic activities [10, 11]. MSC-CM directly contributes to the recovery of the damaged tissues [11]. Considering their regenerative and restorative abilities, MSC-CM from different sources of MSC is proposed as the main biological effector as a possible alternative to MSC treatment in NDD [3, 12]

Methods

Results

Discussion

Conclusion