Selected Kefir Water from Malaysia Attenuates Hydrogen Peroxide-Induced Oxidative Stress by Upregulating Endogenous Antioxidant Levels in SH-SY5Y Neuroblastoma Cells.

Muganti Rajah Kumar,Melati Khalid,Mas Jaffri Masarudin,Swee Keong Yeap,Noorjahan Banu Alitheen,Muhammad Nazirul Mubin Aziz,Janna Ong Abdullah,Han Chung Lee,Adam Thean Chor Leow,Nurul Elyani Mohamad

doi:10.3390/antiox10060940

Abstract

Kefir, a fermented probiotic drink was tested for its potential anti-oxidative, anti-apoptotic, and neuroprotective effects to attenuate cellular oxidative stress on human SH-SY5Y neuroblastoma cells. Here, the antioxidant potentials of the six different kefir water samples were analysed by total phenolic content (TPC), total flavonoid content (TFC), ferric reducing antioxidant power (FRAP), and 2,2′-diphenyl-1-picrylhydrazyl radical (DPPH) assays, whereas the anti-apoptotic activity on hydrogen peroxide (H2O2) induced SH-SY5Y cells was examined using MTT, AO/PI double staining, and PI/Annexin V-FITC assays. The surface and internal morphological features of SH-SY5Y cells were studied using scanning and transmission electron microscopy. The results indicate that Kefir B showed the higher TPC (1.96 ± 0.54 µg GAE/µL), TFC (1.09 ± 0.02 µg CAT eq/µL), FRAP (19.68 ± 0.11 mM FRAP eq/50 µL), and DPPH (0.45 ± 0.06 mg/mL) activities compared to the other kefir samples. The MTT and PI/Annexin V-FITC assays showed that Kefir B pre-treatment at 10 mg/mL for 48 h resulted in greater cytoprotection (97.04%), and a significantly lower percentage of necrotic cells (7.79%), respectively. The Kefir B pre-treatment also resulted in greater protection to cytoplasmic and cytoskeleton inclusion, along with the conservation of the surface morphological features and the overall integrity of SH-SY5Y cells. Our findings indicate that the anti-oxidative, anti-apoptosis, and neuroprotective effects of kefir were mediated via the upregulation of SOD and catalase, as well as the modulation of apoptotic genes (Tp73, Bax, and Bcl-2).

Highlights

Neurodegenerative diseases (NDD) affect millions of people and cause 17.1% of total disability-adjusted life-years for all causes and ages worldwide [1,2]
The antioxidant activity of the kefir water samples were determined through the total phenolic content (TPC), total flavonoid content (TFC), diphenyl-1-picrylhydrazyl radical (DPPH), and ferric reducing antioxidant power (FRAP) assays
The DPPH assay shows that only a 0.45 ± 0.06 mg/mL concentration is required for Kefir B to scavenge 50% of 2,2diphenyl-1-picrylhydrazyl free radical activity, unlike other kefir samples that require higher concentrations

Summary

Introduction

Neurodegenerative diseases (NDD) affect millions of people and cause 17.1% of total disability-adjusted life-years for all causes and ages worldwide [1,2] They are mainly due to the persistence of chronic oxidative stress and neuroinflammation. A low concentration of H2 O2 is a potent mitogen in a number of cell types, beyond the threshold level, its oxidisable effects often result in cell damage and apoptosis, causing human disorders [4,5,6,7] This reaction usually takes place in the brain neuron cells as they are highly sensitive, require high energy, and are niduses for many peroxidisable molecules [8,9]. Several studies revealed that protective factors, such as antioxidants, can prevent H2 O2 -induced oxidative damages [4,5,11,12]

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