Cordycepin attenuates Salivary Hypofunction through the Prevention of Oxidative Stress in Human Submandibular Gland Cells

Abstract

Xerostomia (dry mouth) is a significant age-related condition. Meanwhile, cordycepin, the natural therapeutic agent, has demonstrated an anti-aging effect. Therefore, the present study aimed to investigate the preventive effects of cordycepin on secretory function in an in vitro model of hydrogen peroxide (H 2 O 2 )-induced salivary hypofunction. After being exposed to H 2 O 2 , human submandibular gland (HSG) cells were treated with various concentrations of cordycepin (6.25-50 µM) for 24, 48, and 72h. To evaluate cell proliferation and reactive oxygen species (ROS) generation, 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide and 2, 7'-dichlorodihydrofluorescein diacetate assays were performed. The amylase activity was kinetically measured by 2-chloro- p -nitrophenol linked with maltotrioside. The expression of salivary, antioxidant and apoptotic markers at mRNA and protein levels were performed by reverse transcriptase polymerase chain reaction (RT-PCR) and immunofluorescence analysis, respectively. We demonstrated that cordycepin (6.25-25 µM) contributed to significant increases in expression of the salivary marker genes, alpha-amylase 1 (AMY1A) and aquaporin-5 (AQP5), and in amylase secretion without changes in cell viability. Under oxidative stress, HSG cells showed remarkable dysfunction. Cordycepin rescued the protective effects partially by decreasing ROS generation and restoring the expression of the salivary proteins, AMY and AQP5 via anti-oxidant and anti-apoptotic activity. In addition, the amount of amylase that was secreted from HSG cells cultured in cordycepin was increased. In conclusion, cordycepin demonstrated a protective effect on H 2 O 2 - induced HSG cells by decreasing ROS generation and upregulating the salivary function markers, AMY1A and AQP5 , at both the transcriptional and translational levels.

A Systematic Review of the Biological Effects of Cordycepin

“…We noted 45 papers claiming that the cytotoxic effect of cordycepin at bioactive concentrations was low. We noted the highest concentration which promoted cell survival or did not cause statistically significant cell death in these papers and determined the median and the range [ 28 , 29 , 30 , 32 , 33 , 34 , 35 , 36 , 37 , 38 , 39 , 40 , 41 , 42 , 43 , 44 , 69 , 70 , 80 , 95 , 98 , 102 , 105 , 106 , 117 , 142 , 145 , 164 , 184 , 240 , 270 , 301 , 315 , 317 , 326 , 327 , 328 , 329 , 330 , 331 , 332 , 333 , 334 , 335 , 336 ].…”

Effects of polyphenols in non-centrifugal cane sugar on saliva secretion: <i>in vitro</i> and <i>in vivo</i> experiments and a randomized controlled trial

“…AQP5 is a water channel protein located in salivary gland acinar cells that mediates transcellular water movement. (32)(33)(34) Muscarinic receptor stimulation also induces translocation of AQP5 from the cytoplasm to the apical side of the acinar cells. (35) Previous studies reported that an antioxidant found in green tea and some plant-derived compounds promote salivation via AQP5 translocation.…”

Apigenin, a Single Active Component of Herbal Extract, Alleviates Xerostomia via ERα-Mediated Upregulation of AQP5 Activation

“…Real-Time Quantitative PCR HSG cells (Jaiboonma et al, 2020) were seeded in 6-well plates with a DMEM containing 1% FBS. The cells were stimulated with apigenin (0.01, 0.1, and 1 μM) and E2 (0.1 μM) for 48 h. The total RNA from differently treated cells was extracted using a total RNA extraction reagent.…”