Anti-Melanogenesis Effect of Eucheuma cottonii Extract

in B16F10 Murine Melan-a Melanocytes

Monica Drestia
2011-080-074

Faculty of Biotechnology
Atma Jaya Catholic University of Indonesia
Jakarta
2014
Introduction
Introduction
(Costin & Hearing 2007)
 (Selvan et al. 2014)

Nutrients (per 100 g) Concentration (g)

Carbohydrates 57.3
Proteins 4.5
Eucheuma cottonii Fat 0.89
Ash 28.9

(Abirami & Kowsalya 2011)

 Methods
E. cottonii Extraction and Cell Cultivation

Melanogenesis Induction & E. cottonii Extract

Treatment

Py-GC/MS

RNA Isolation & qRT-PCR

Cytotoxicity Assay

Tyrosinase Activity Assay

E. cottonii Extraction

+ → →

(Hindritiani et al. 2013) ←

Cell Cultivation

→
Melanogenesis Induction

E. cottonii Extract Treatment

(Lee et al. 2010)

 Py-GC/MS

Approximately, 0.5 g of
sample was injected to the
capillary column.

Pyrolysis temperature was set

to 280oC using helium as a
carrier gas.

(Zayed et al. 2014)

Cytotoxic Assay

37oC
3 hr 590 nm
+ →
MTT reagent
Treated cells (20 µL)
(Hindritiani et al. 2013)
Measurement of Tyrosinase Activity

37oC
1 hr
→ →
475 nm

Treated cells,
lysed
+ L-DOPA
RNA Extraction & qRT-PCR

→
+ TRIzol

treated cells
Kappa SYBR FAST One-Step qRT-PCR
Primer   Sequences (5’→3’) Size (bp)
MITF Forward CAGCCAACCTTCCCAACATA 300
  Reverse CCAGCTTCTTCTGTCGGTTT  
GAPDH Forward CTTGAGCTAGGACTGGATAAGC 296
  Reverse TGTAGTTGAGGTCAATGAAGGG  
 Oleic acid
Linoleic acid
Results

Phytol
 Results

MITF gene expression of ethanolic E. cottonii extract in PMA-induced

B16F10 cells after 24 hour of incubation
 Results

Tyrosinase activity of ethanolic E. cottonii extract on PMA-induced B16F10 cells.

 Results

Cytotoxic activity of ethanolic E. cottonii extract on PMA-induced B16F10 cells viability

after 24 hour of incubation.
 Discussion
Py-GC/MS

Fatty acids and sterols are widely distributed in seaweeds (Rajasulochana et al. 2010).

Fatty acids cause a stimulation of tyrosinase ubiquitination and degradation by the

proteasome, which leads to a decrease of active tyrosinase amount inside the
melanocytes (Solano et al. 2006).

Vitamin A gives benefit to skin cells through regulation of epidermal cell growth and
maintenance of normal cell (Fu et al. 2007).
 Discussion
qRT-PCR
Analysis

Fatty acids and sterols are widely distributed in seaweeds (Rajasulochana et al. 2010).

Fatty acids cause a stimulation of tyrosinase ubiquitination and degradation by the

proteasome, which leads to a decrease of active tyrosinase amount inside the
melanocytes (Solano et al. 2006).

Vitamin A gives benefit to skin cells through regulation of epidermal cell growth and
maintenance of normal cell (Fu et al. 2007).
 E. cottonii extract decreased the level of MITF expression in a dose-independent manner. The
optimum concentration of E. cottonii extract with the lowest MITF expression (about 0.11%) was 100
ug mL-1.
 It was reported that marine macroalgae polysaccharides (such as carrageenan and fucoidan)
significantly decreased MITF expression of melanocytes. MITF is an important regulating factor of
melanogenesis. Inhibition of MITF expression can reduce tyrosinase activity and inhibit melanin
formation of the B16F10 cells (Song et al. 2015).
 Conclusion

• Ethanolic E. cottonii extract up to concentration 10 µg mL-1

suppressed MITF gene expression and tyrosinase activity in
PMA-induced B16F10 murine melan-a melanocytes without
any apparent significant cytotoxic activity.
• The composition of E. cottonii extract including
polysaccharide, fatty acids, and secondary metabolite
components.
• E. cottonii extract can be considered as natural and safe
skin whitening agent
 References
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