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Cited by 112Skin Protection and AgingPlant tissue culture and regenerationMedicinal Plant Research

The DOI matches its Crossref record; 60 of the 60 references it cites were found in a registry or an index.

Insights into antioxidant activities and anti-skin-aging potential of callus extract from Centella asiatica (L.)

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Buranasudja, Visarut, Rani, Dolly, Malla, Ashwini, Kobtrakul, Khwanlada, Vimolmangkang, Sornkanok·2021·Scientific Reports
DOI 10.1038/s41598-021-92958-7PMC PMC8241881
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Abstract

Formation of oxidative stress in dermal fibroblasts plays crucial roles in aging processes of skin. The use of phytochemicals that can promote capacity of fibroblasts to combat oxidative stress is an attractive strategy to prevent skin aging and promote skin beauty. Centella asiatica has been used to treat multitude of diseases for centuries. Previous investigations demonstrated that extracts from C. asiatica have a broad range of beneficial activities through their antioxidant activity. Hence, the extract from this medicinal plant could be a great candidate for anti-skin-aging agent. Callus culture offers a powerful platform for sustainable, rapid and large-scale production of phytochemicals to serve extensive demands of pharmaceutical and cosmeceutical industries. Here, we demonstrated the application of callus culture of Centella asiatica to produce bioactive metabolites. The 50% ethanolic extract of callus culture has distinctive features of chemical compositions and biological profiles. Information from HPTLC-DPPH and HPLC analysis suggested that the callus extract comprises distinctive antioxidant compounds, compared with those isolated from authentic plant. Moreover, results from cell culture experiment demonstrated that callus extract possesses promising antioxidant and anti-skin-aging activities. Pre-treatment with callus extract attenuated H 2 O 2-induced-cytotoxicity on human dermal fibroblasts. The results from RT-qPCR clearly suggested that the upregulation of cellular antioxidant enzymes appeared to be major contributor for the protective effects of callus extract against oxidative stress. Moreover, supplementation with callus extract inhibited induction of matrix metalloprotease-9 following H 2 O 2 exposure, suggesting its potential anti-skin-aging activity. Our results demonstrate the potential utility of C. asiatica callus extract as anti-skin-aging agent in cosmeceutical preparations.

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Figure 1 The antioxidant activities of Centella extracts are not due to asiaticoside, asiatic acid, madecassoside and madecassic acid. HPTLC fingerprints of triterpene standards (1, madecassoside; 2, asiaticoside; 3 madecassic acid; 4, asiatic acid), APE (5, methanol; 6, 95 % ethanol; 7, 70 % ethanol; 8, 50 % ethanol), and CE (9, methanol; 10, 95 % ethanol; 11, 70 % ethanol; 12, 50 % ethanol). (A) The HPTLC plate was derivatized with anisaldehyde sulphuric acid reagent and visualized with white light. (B) The HPTLC plate was dipped with DPPH reagent and visualized with white light.
Figure 1 The antioxidant activities of Centella extracts are not due to asiaticoside, asiatic acid, madecassoside and madecassic acid. HPTLC fingerprints of triterpene standards (1, madecassoside; 2, asiaticoside; 3 madecassic acid; 4, asiatic acid), APE (5, methanol; 6, 95 % ethanol; 7, 70 % ethanol; 8, 50 % ethanol), and CE (9, methanol; 10, 95 % ethanol; 11, 70 % ethanol; 12, 50 % ethanol). (A) The HPTLC plate was derivatized with anisaldehyde sulphuric acid reagent and visualized with white light. (B) The HPTLC plate was dipped with DPPH reagent and visualized with white light.CC BY · PubMed Central
Figure 2 The antioxidant activities of Centella extracts are not due to kaempferol, quercetin and rutin. HPTLC fingerprints of flavonoid standards (1, kaempferol; 2, quercetin; 3, rutin), APE (4, methanol; 5, 95 % ethanol; 6, 70 % ethanol; 7, 50 % ethanol), and CE (8, methanol; 9, 95 % ethanol; 10, 70 % ethanol; 11, 50 % ethanol). (A) The HPTLC plate derivatized with NP/PEG solution and visualized under UV light at 366 nm. (B) The HPTLC plate was immersed with DPPH reagent and visualized with white light.
Figure 2 The antioxidant activities of Centella extracts are not due to kaempferol, quercetin and rutin. HPTLC fingerprints of flavonoid standards (1, kaempferol; 2, quercetin; 3, rutin), APE (4, methanol; 5, 95 % ethanol; 6, 70 % ethanol; 7, 50 % ethanol), and CE (8, methanol; 9, 95 % ethanol; 10, 70 % ethanol; 11, 50 % ethanol). (A) The HPTLC plate derivatized with NP/PEG solution and visualized under UV light at 366 nm. (B) The HPTLC plate was immersed with DPPH reagent and visualized with white light.CC BY · PubMed Central
Figure 3 Centella extract from callus culture has unique chemical profiles. HPLC chromatogram of (A) standard solutions of triterpenes and flavonoids (1, rutin; 2, madecassoside; 3, asiaticoside; 4, quercetin; 5, kaempferol; 6, madecassic acid; 7, asiatic acid); (B–E) Authentic plant extract (B, methanol; C, 95% ethanol; D, 70% ethanol, E, 50% ethanol); and (F–I) Callus extract (F, methanol; G, 95% ethanol; H, 70% ethanol, I, 50% ethanol). The retention time (RT) was shown in parenthesis.
Figure 3 Centella extract from callus culture has unique chemical profiles. HPLC chromatogram of (A) standard solutions of triterpenes and flavonoids (1, rutin; 2, madecassoside; 3, asiaticoside; 4, quercetin; 5, kaempferol; 6, madecassic acid; 7, asiatic acid); (B–E) Authentic plant extract (B, methanol; C, 95% ethanol; D, 70% ethanol, E, 50% ethanol); and (F–I) Callus extract (F, methanol; G, 95% ethanol; H, 70% ethanol, I, 50% ethanol). The retention time (RT) was shown in parenthesis.CC BY · PubMed Central
Figure 4 Centella extracts prevent H2O2-induced cytotoxicity in dermal fibroblasts. (A) and (B) CE and APE at 15–60 µg/mL did not cause toxicity in dermal fibroblasts. BJ cells were incubated with CE or APE (15–60 µg/mL) for 24 h. (C) and (D) CE and APE inhibit oxidative damage from H2O2 exposure. BJ cells were pre-incubated with CE or APE (15–60 µg/mL) for 24 h then exposed to H2O2 (200 µM) for 1 h. The cell viability was evaluated with MTT assay immediately after the treatments. (E) and (F) CE inhibit depletion of intracellular ATP following H2O2 exposure. The protocol for treatment was as (A) and (C). The amounts of ATP were measured immediately after treatments (n = 3; mean ± SEM; *P < 0.05 vs untreated control; †P < 0.05 vs H2O2-treated cells).
Figure 4 Centella extracts prevent H2O2-induced cytotoxicity in dermal fibroblasts. (A) and (B) CE and APE at 15–60 µg/mL did not cause toxicity in dermal fibroblasts. BJ cells were incubated with CE or APE (15–60 µg/mL) for 24 h. (C) and (D) CE and APE inhibit oxidative damage from H2O2 exposure. BJ cells were pre-incubated with CE or APE (15–60 µg/mL) for 24 h then exposed to H2O2 (200 µM) for 1 h. The cell viability was evaluated with MTT assay immediately after the treatments. (E) and (F) CE inhibit depletion of intracellular ATP following H2O2 exposure. The protocol for treatment was as (A) and (C). The amounts of ATP were measured immediately after treatments (n = 3; mean ± SEM; *P < 0.05 vs untreated control; †P < 0.05 vs H2O2-treated cells).CC BY · PubMed Central
Figure 5 Centella extracts elevate transcription of antioxidant enzymes. (A–D) CE promoted CAT expression, while APE enhanced SOD1 and SOD2 expression. BJ cells were incubated with CE (15–60 µg/mL; 24 h) or APE (60 µg/mL; 24 h) or H2O2 (200 µM; 1 h). The expressions of antioxidant machineries, including CAT, GPx1, SOD1, and SOD2, were determined with RT-PCR following treatments (n = 3; mean ± SEM; *P < 0.05 vs untreated control).
Figure 5 Centella extracts elevate transcription of antioxidant enzymes. (A–D) CE promoted CAT expression, while APE enhanced SOD1 and SOD2 expression. BJ cells were incubated with CE (15–60 µg/mL; 24 h) or APE (60 µg/mL; 24 h) or H2O2 (200 µM; 1 h). The expressions of antioxidant machineries, including CAT, GPx1, SOD1, and SOD2, were determined with RT-PCR following treatments (n = 3; mean ± SEM; *P < 0.05 vs untreated control).CC BY · PubMed Central
Figure 6 Centella extracts promote expression of antioxidant enzymes following H2O2 treatment. (A–D) In response to H2O2 treatment, CAT, GPX1 and SOD1 expressions were increased in CE-treated fibroblasts, while SOD2 transcription were elevated in APE-treated fibroblasts. BJ cells were incubated with CE (60 µg/mL; 24 h) or APE (60 µg/mL; 24 h), then exposed to H2O2 (200 µM; 1 h). The transcriptional levels of CAT, GPx1, SOD1, and SOD2, were evaluated with RT-PCR following treatments (n = 3; mean ± SEM; †P < 0.05 vs H2O2-treated cells).
Figure 6 Centella extracts promote expression of antioxidant enzymes following H2O2 treatment. (A–D) In response to H2O2 treatment, CAT, GPX1 and SOD1 expressions were increased in CE-treated fibroblasts, while SOD2 transcription were elevated in APE-treated fibroblasts. BJ cells were incubated with CE (60 µg/mL; 24 h) or APE (60 µg/mL; 24 h), then exposed to H2O2 (200 µM; 1 h). The transcriptional levels of CAT, GPx1, SOD1, and SOD2, were evaluated with RT-PCR following treatments (n = 3; mean ± SEM; †P < 0.05 vs H2O2-treated cells).CC BY · PubMed Central
Figure 7 Centella extracts inhibit expression of MMP9 following H2O2 treatment. (A) Exposure to H2O2 resulted in elevation of MMP9 expression, while CE and APE did not affect the transcription of MMP9. BJ cells were treated CE (15–60 µg/mL; 24 h) or APE (60 µg/mL; 24 h) or H2O2 (200 µM; 1 h). (B) CE and APE suppressed H2O2-induced MMP9 expression. BJ cells were pre-incubated with CE or APE (60 µg/mL) for 24 h then exposed to H2O2 (200 µM) for 1 h. The transcriptional levels of MMP9 were evaluated with RT-PCR following treatments (n = 3; mean ± SEM; *P < 0.05 vs untreated control; †P < 0.05 vs H2O2-treated cells).
Figure 7 Centella extracts inhibit expression of MMP9 following H2O2 treatment. (A) Exposure to H2O2 resulted in elevation of MMP9 expression, while CE and APE did not affect the transcription of MMP9. BJ cells were treated CE (15–60 µg/mL; 24 h) or APE (60 µg/mL; 24 h) or H2O2 (200 µM; 1 h). (B) CE and APE suppressed H2O2-induced MMP9 expression. BJ cells were pre-incubated with CE or APE (60 µg/mL) for 24 h then exposed to H2O2 (200 µM) for 1 h. The transcriptional levels of MMP9 were evaluated with RT-PCR following treatments (n = 3; mean ± SEM; *P < 0.05 vs untreated control; †P < 0.05 vs H2O2-treated cells).CC BY · PubMed Central
Figure 8 Proposed mechanisms for antioxidant and anti-skin aging activities of Centella extract. Pre-treatment with Centella extracts prevent H2O2-induced cytotoxicity on dermal fibroblasts by upregulation of cellular antioxidant machineries. Supplementation with CE upregulated CAT, GPx1 and SOD1 expression, whereas pre-treatment with APE induced SOD2 expression following H2O2 treatment. Moreover, pre-treatment with CE or APE suppressed H2O2-mediated-upregulation of MMP-9. This figure was created with BioRender.com.
Figure 8 Proposed mechanisms for antioxidant and anti-skin aging activities of Centella extract. Pre-treatment with Centella extracts prevent H2O2-induced cytotoxicity on dermal fibroblasts by upregulation of cellular antioxidant machineries. Supplementation with CE upregulated CAT, GPx1 and SOD1 expression, whereas pre-treatment with APE induced SOD2 expression following H2O2 treatment. Moreover, pre-treatment with CE or APE suppressed H2O2-mediated-upregulation of MMP-9. This figure was created with BioRender.com.CC BY · PubMed Central

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Metadata

Authors
Buranasudja, V., Rani, D., Malla, A., Kobtrakul, K., & Vimolmangkang, S.
Affiliations
Chulalongkorn University (TH)
Corresponding author
Vimolmangkang, Sornkanok
ORCID
Venue
Scientific Reports
Publisher
Springer Science and Business Media LLC
Published
2021-06-29
Volume
11
Issue
1
Pages
13459-13459
ISSN
2045-2322 (ISSN-L)
DOI
10.1038/s41598-021-92958-7
PMC
PMC8241881
OpenAlex
W3135110362
Cited by
112
References
60
Citations by year
2026: 24 · 2025: 24 · 2024: 27 · 2023: 22 · 2022: 13 · 2021: 2
FWCI
14.21
Type
journal article
Language
en
Open access
Licence
Locations
  • Scientific Reports (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
Funders
  • Rachadapisek Sompote Fund for Postdoctoral Fellowships, Graduate School of Chulalongkorn University
  • Postdoctoral fellowship, the Second Century Fund (C2F), Chulalongkorn University
  • PMU-C of the Office of the National Higher Education Science Research and Innovation Policy Council, Thailand
Topics
  • Health Sciences › Medicine › Dermatology › Skin Protection and Aging
  • Life Sciences › Agricultural and Biological Sciences › Plant Science › Plant tissue culture and regeneration
  • Life Sciences › Agricultural and Biological Sciences › Plant Science › Medicinal Plant Research
Keywords
Centella asiatica, callus culture, antioxidant activity, oxidative stress, dermal fibroblasts, hydrogen peroxide cytotoxicity, antioxidant enzymes, MMP-9, cosmeceuticals, phytochemicals, ethanol extract
MeSH terms
Antioxidants, Cells, Cultured, Fibroblasts, Humans, Plant Extracts, Triterpenes, Skin Aging, Centella

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