The Protective Effect of Polyherbal Formulation, Harak Formula, on UVA-Induced Photoaging of Human Dermal Fibroblasts and Mouse Skin via Promoting Nrf2-Regulated Antioxidant Defense

Abstract

Polyherbal formulation combining multiple herbs is suggested to achieve enhanced therapeutic effects and reduce toxicity. Harak herbal formula (HRF) extracts were proposed to regulate skin responses to UVR through their ability to suppress UVA-induced matrix metalloproteinase-1 (MMP-1) and pigmentation via promoting antioxidant defenses in in vitro models. Therefore, natural products targeting Nrf2 (nuclear factor erythroid 2-related factor 2)-regulated antioxidant response might represent promising anti-photoaging candidates. Hesperetin (HSP) was suggested as a putative bioactive compound of the HRF, as previously shown by its chemical profiling using the liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (LC-QTOF-MS). In this study, we explored the anti-photoaging effects of HRF extracts and HSP on normal human dermal fibroblasts (NHDFs) and mouse skin exposed to UVA irradiation. Pretreatment of NHDFs with HRF extracts and HSP protected against UVA (8 J/cm2)-mediated cytotoxicity and reactive oxygen species (ROS) formation. The HRF and HSP pretreatment also attenuated the UVA-induced MMP-1 activity and collagen depletion concomitant with an upregulation of Nrf2 activity and its downstream genes (GST and NQO-1). Moreover, our findings provided the in vivo relevance to the in vitro anti-photoaging effects of HRF as topical application of the extracts (10, 30 and 100 mg/cm2) and HSP (0.3, 1, and 3 mg/cm2) 1 h before UVA exposure 3 times per week for 2 weeks (a total dose of 60 J/cm2) mitigated MMP-1 upregulation, collagen loss in correlation with enhanced Nrf2 nuclear accumulation and its target protein GST and NQO-1 as well as reduced 8-hydroxy-2′-deoxyguanosine (8-OHdG) in irradiated mouse skin. Thus, our findings revealed that HRF extracts and HSP attenuated UVA-induced photoaging via upregulating Nrf2, together with their abilities to reduce ROS formation and oxidative damage. Our study concluded that the HRF and its bioactive ingredient HSP may represent potential candidates for preventing UVA-induced photoaging via restoration of redox balance.

Role of Phytochemicals in Skin Photoprotection via Regulation of Nrf2

“…Furthermore, the analysis of proteome profiles revealed that rutin treatment caused an induction of proteins involved in the antioxidant defenses and a reduction of proteins involved in the degradation of Nrf2 in UVB-irradiated dermal fibroblasts ( Gegotek et al, 2018 ). Hesperidin, (hesperetin-7-rutinoside), and its aglycone, hesperetin, mostly found in citrus fruits and botanical drugs, have been demonstrated to provide in vitro and in vivo anti-photoaging effects on the skin via stimulating collagen synthesis in association with the antioxidant properties ( Lohakul et al, 2021a ). Citrus sinensis peel extract containing hesperidin loaded lipid nanoparticles showed photoprotective effects on UVR-mediated induction of MMP-1 via JNK signaling, reduction of collagen accompanied by decreased SOD protein production as well as stimulated inflammatory markers (COX-2, prostaglandin E2) and lipid oxidation product, malondialdehyde (MDA), in mouse skin ( Amer et al, 2021 ).…”

“…In addition, a clinical trial showed that a 12-week topical application of a serum containing 0.1% hesperetin significantly promoted skin hydration and elasticity via enhancing the synthesis of hyaluronic acid ( Sheen et al, 2021 ). Moreover, hesperetin and polyherbal formula extracts containing hesperetin topically applied to mouse skin before UVA exposure three times per week for 2 weeks (a total dose of 60 J/cm 2 ) significantly attenuated MMP-1 upregulation and collagen depletion concomitant with promoting Nrf2 activity and the level of its target proteins (GST and NQO-1) as well as reducing 8-hydroxy-2′-deoxyguanosine (8-OHdG), a product of oxidatively damaged DNA damage, in irradiated mouse skin ( Lohakul et al, 2021a ). Grape fruit stem extract from Muscat Bailey A containing catechin, epicatechin and trans-resveratrol showed protective effects on UVB-induced destruction of collagen fiber through reduction of MMP-1 expression in association with a decrease in lipid peroxidation and restoration of GSH levels in mouse skin ( Cho et al, 2018 ).…”

Flavonoids Present in Propolis in the Battle against Photoaging and Psoriasis

“…The group of the flavanones have the C ring saturated and also are named dihydroflavones because the only difference with the flavones is that the double bond between positions 2 and 3 is saturated [ 49 ]. In 2021, Lohakul et al [ 82 ] reported that hesperetin pre-administration (2.5, 5, or 10 μg/mL) in UVA-irradiated HDFs decreased ROS generation and also reduced collagen depletion and MMP-1 activity. Additionally, this flavonoid positively regulated Nrf2 activity and its genes NAD(P)H: quinone oxidoreductase 1 (NQO-1) and glutathione S-transferase (GST) ( Table 3 ).…”

“…Furthermore, this flavonoid increased nuclear Nrf2 levels and its NQO-1 and GST target proteins. Additionally, the hesperetin decreased the 8-OHdG, an indicator of oxidative DNA damage, in mice skin, indicating an attenuation of photoaging through the ability of hesperetin to positively regulate Nrf2; therefore, this flavonoid could be a promising agent to restore redox equilibrium and help in treatment against skin photoaging [ 82 ].…”

Hesperetin from Root Extract of Clerodendrum petasites S. Moore Inhibits SARS-CoV-2 Spike Protein S1 Subunit-Induced NLRP3 Inflammasome in A549 Lung Cells via Modulation of the Akt/MAPK/AP-1 Pathway

“…It is widely grown in India, Malaysia, and Thailand. In Thai traditional medicine, its leaves and roots are used in the treatment of fever, inflammation, skin diseases, and asthma [ 15 , 16 ]. In addition, C. petasites has generally been used in formulations of some multi-herb medicinal recipes such as the traditional formula “Ha-Rak” remedy (also known as Ben-Cha-Lo-Ka-Wi-Chian).…”