Cathelicidin-NV from Nanorana ventripunctata effectively protects HaCaT cells, ameliorating ultraviolet B-induced skin photoaging.

Cathelicidins are diverse effector molecules in the vertebrate immune system and are related to immune regulation, inflammatory response, wound healing, and blood vessel formation. However, little is known about their free radical scavenging ability, especially in vivo. In this study, a cathelicidin molecule (cathelicidin-NV, ARGKKECKDDRCRLLMKRGSFSYV) previously identified from the spot-bellied plateau frog (Nanorana ventripunctata) (Anura, Dicroglossidae, Dicroglossinae) by us was shown to alleviate ultraviolet B (UVB)-induced skin photoaging in mice. Cathelicidin-NV effectively suppressed cytotoxicity, DNA fragmentation, apoptosis and reduced the protein expression levels of JNK, c-Jun, and MMP-1, which are involved in the regulation of collagen degradation in HaCaT cells induced by UVB irradiation. Furthermore, cathelicidin-NV also scavenged UVB-induced intracellular reactive oxygen species (ROS). Taken together, cathelicidin-NV directly scavenged excessive intracellular ROS to protect HaCaT cells, and subsequently alleviated UVB-induced skin photoaging. This study extends reports on the antioxidant function of the cathelicidin family. In addition, the properties of cathelicidin-NV make it an excellent candidate for the prevention and treatment of UV-induced skin photoaging.

A Frog Peptide Ameliorates Skin Photoaging Through Scavenging Reactive Oxygen Species.

Although many bioactive peptides have been identified from the frog skins, their protective effects and the molecular mechanisms against skin photodamage are still poorly understood. In this study, a novel 20-residue peptide (antioxidin-NV, GWANTLKNVAGGLCKMTGAA) was characterized from the skin of plateau frog Nanorana ventripunctata. Antioxidin-NV obviously decreased skin erythema, thickness and wrinkle formation induced by Ultraviolet (UV) B exposure in hairless mice. In UVB-irradiated keratinocytes (HaCaT cells) and hairless mice, it effectively inhibited DNA damage through reducing p-Histone H2A.X (γH2AX) expression, alleviated cell apoptosis by decreasing the expression of apoptosis-specific protein (cleaved caspase 3), and reduced interleukin-6 (IL-6) production via blocking UVB-activated Toll-like receptor 4 (TLR4)/p38/JNK/NF-κB signaling. In UVB-irradiated human skin fibroblasts (HSF cells) and hairless mice, it effectively restored HSF cells survival rate, and rescued α-SMA accumulation and collagen (especially type I collagen) production by restoring transforming growth factor-ß1 (TGF-ß1)/Smad2 signaling. We found that antioxidin-NV directly and rapidly scavenged intracellular and mitochondrial ROS in HaCaT cells upon UVB irradiation, and quickly eliminated the artificial free radicals, 2, 2'-azinobis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS+). Taken together, antioxidin-NV directly and rapidly scavenged excessive ROS upon UVB irradiation, subsequently alleviated UVB-induced DNA damage, cell apoptosis, and inflammatory response, thus protecting against UVB-induced skin photoaging. These properties makes antioxidin-NV an excellent candidate for the development of novel anti-photoaging agent.

Direct sunlight exposure reduces hair cortisol levels in rhesus monkeys (Macaca mulatta).

Major depressive disorder (MDD), commonly known as depression, is a mental disease characterized by a core symptom of low mood. It lasts at least two weeks (Badamasi et al., 2019; Wang et al., 2019) and is frequently accompanied by low self-esteem, loss of interest in routinely enjoyable activities, low energy, and unexplained pain (Huey et al., 2018; Park et al., 2012; Post & Warden, 2018; Rice et al., 2019; Xiao et al., 2018). Approximately 2%-8% of adults with MDD commit suicide (Richards & O'Hara, 2014; Strakowski & Nelson, 2015), and around half of suicidal individuals suffer depression or other mood disorders (Bachmann, 2018).

The first anionic defensin from amphibians.

A variety of antimicrobial peptides against infection have been identified from the skin of amphibians. However, knowledge on amphibian defensins is very limited. A novel anionic defensin designated PopuDef was purified from the skin of tree frog Polypedates puerensis, and the cDNA encoding PopuDef precursor was cloned from the skin cDNA library. The amino acid sequence of PopuDef (net charge: -2, pI: 4.75) shared the highest identity of 57 % (25/44) with the salamander defensin CFBD-1 (net charge: 0, pI: 6.14) from urodela amphibians. PopuDef showed moderate antimicrobial activities against P. aeruginosa and S. aureus (MICs are 19.41 and 17.25 µM, respectively), and relatively weak activities against E. coli and B. subtilis (MICs are 38.82 and 43.14 µM, respectively). Tissue distribution analysis indicated that relatively high expression level of PopuDef mRNA was observed in immune-related tissues including skin, gut, lung and spleen. Furthermore, the expression level of PopuDef was significantly upregulated in these tissues after tree frogs were infected with different bacteria strains mentioned above. Interestingly, the induction of PopuDef challenged with E. coli or B. subtilis, which was less sensitive to PopuDef, was much higher than that did with P. aeruginosa or S. aureus. These findings highlight the key role of PopuDef in innate immunity against infection. To our knowledge, PopuDef is the first anionic defensin characterized from amphibians.

Molecular cloning and functional characterization of novel antimicrobial peptides from the skin of brown frog, Rana zhenhaiensis.

Rana zhenhaiensis, a species of brown frog, is widely distributed in central and south China. In the present study, a total of 14 cDNA sequences encoding eight novel antimicrobial peptides (AMPs) were cloned from the synthesized cDNAs of R. zhenhaiensis skin. The eight novel AMPs belong to four families: brevinin-1 (four peptides), brevinin-2 (one peptide), ranatuerin-2 (one peptide) and chensinin-1 (two peptides), five AMPs from the four families (brevinin-1ZHa, brevinin-1ZHb, brevinin-2ZHa, ranatuerin-2ZHa and chensinin-1ZHa) were chemically synthesized, their antimicrobial and hemolytic activities were examined. The results indicated that the five AMPs possess different antimicrobial and hemolytic activities. Of these, brevinin-2ZHa exhibited the strongest and most broad-spectrum antimicrobial activity. Furthermore, scanning electron microscopy (SEM) experiment was carried out to investigate the potential antimicrobial mechanism of chensinin-1ZHa. The result indicated that chensinin-1ZHa may exert its function through disruption of the bacterial membrane.