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Effect of Moderate Exercise on Breast Milk Leukocytes in Exclusively
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Nano Capsulated Polyphenol Extracted from Oyster Mushroom (Pleurotus
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Khaled F. Mahmoud; Azza A. Amin; Effat I. Seliem; Manal F. Salama
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Broccoli Flower Extract (Brassica oleracea L. var.italica Plenck) Inhibits Photoaging
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Nelva K. Jusuf
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Effect of Kt6 Variant Cowpea (Vigna Unguiculata) Extract on Matrix
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Nisful Laila Sa’adah, Nanda Wahyu Anandita
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Antiarthritic Activity of Pacar Air (Impatiens balsamina Linn.) Herb Extract in
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Clinicopathological changes in equine herpes virus type 1 (EHV-1) infection in
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Fararh K.M., Kandil O.M., Abd-Allah O.A. and Thabet N.F.
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Determination of Antioxidant activity by High Performance Liquid
Chromatography, Phenolic and Flavonoid contents of Vincetoxicum nigrum
Sepideh Nourian, Ali Mohammadi Sani, Ebrahim Golmakani, Peyman Feizi,
Katayoun Roghani
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Ginkgo Biloba Extract Effect on Oxidative Stress Marker Malonildialdehyde, Redox
Enzyme Gluthation Peroxidase, Visual Field Damage, and Retinal Nerve Fiber
Layer Thickness in Primary Open Angle Glaucoma
MasithaDewiSari, Aslim D Sihotang, Aznan Lelo
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Antidiabetic Activity of Ipomoea batatas L. Leaves Extract In
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Nasdiwaty Daud, Rosidah, M Pandapotan Nasution
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Stability Indicating High Performance Liquid Chromatographic Method for the
Estimation of Carisoprodol in Bulk and in Tablet Dosage form
D. Murali and C. Rambabu
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Anthocyanin Extraction from Purple Sweet Potato Cultivar Antin-3 (Ipomoea
batatas L.) using Maceration, Microwave Assisted Extraction, Ultrasonic Assisted
Extraction and Their Application as Anti-Hyperglycemic Agents in Alloxan-Induced
Wistar Rats
Luqman Agung Wicaksono, Yunianta, and Tri Dewanti Widyaningsih
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The Effect of Calcium to The Absorption Lead In Male Mice (Mus musculus L.)
Syarifah Nadia, Jansen Silalahi, Muchlisyam
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Span 60 as Surfactant of Topical Microemulsion of Purple Sweet Potato (Ipomoea
batatas L.) Ethanol Extract and Antioxidant Activity Test using Dpph Method
Rise Desnita, Sri Wahdaningsih, Sally Hervianti
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Analysis of Quantitative Structure-Activity Relationship, Pharmacophore, and
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Nur Syamsi Dhuha, Daryono Hadi Tjahjono
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Conventional and molecular approaches in bacterial contamination detection for
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Abdalnabi Jwaied Abid
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Expression of RNA encode FAMeT in mandibular organ of mud crabs Scylla
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Akbar Marzuki Tahya, Muhammad Zairin Junior, Arief Boediono, I Made Artika,
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A Review on Cancer Screening
Malarkodi Velraj, Dhulipalla Sowmya
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Topical Microemulsion’s Formulation of Purple Sweet Potato (Ipomoea batatas
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Ethanol Extract as Antioxidant by using Various Concentration of Span 80
Rise Desnita, Maria Veronika, Sri Wahdaningsih
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Polymerase chain reaction compared with wet mount for detection of Trichomonas
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Hayam khalis Al-Masoudi
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Design and Development of Miglitol Loaded PLGA Polymeric Naoparticles
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The Effect of Simplex Nanoparticles of Vernonia amygdalina Del. on Lipid Profile
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Ardiani Rani, Rosidah, Karsono
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Food Consumption and Body Weight in Mice Treated with Palm Oil
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Ahmed Atia, Nadia Alrawaiq and Azman Abdullah
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Effect of Aluminum-Containing Antacid on Sperm Parameters And testicular Structure in
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Abdul-Hadi Abbas Hadi, Haider Salih Jaffat
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Evaluating the Superoxide Dismutase-1 status in Wild Type and Mutant at codons 12 and
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Moaed E. Al-Gazally, Alaa S. Al-Awad, Hamzah H. Kzar
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Assessment of antioxidant status in different genotypes/phenotypes at codon 72 of TP53
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Moaed E. Al-Gazally, Alaa S. Al-Awad, Hamzah H. Kzar
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Healing Effect of Alginate Liquid against HCl-Induced Gastric Mucosal Lesions in Rats
Anayanti Arianto and Hakim Bangun
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Adverse Effects of Microwaves
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Studies on Diuretics and Laxative Activity of the Hibiscus Tiliaceus Linn. Bark Extracts
Vijay D. Tambe and Rajendra S. Bhambar
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Cognitive and Anti Oxidant Property of Mimusops elengi Linn. in the Experimental
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Chitra V, Tamilanban T, ,Manasa K , Chitra K
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Evaluating Hair Growth Activity of Herbal Hair Oil
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Effect of Allium sativum paste against Antimicrobial, Antioxidant and Cytotoxicity
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A Review on a few medicinal plants possessing anticancer activity against human breast
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Jaikumar B, Jasmine R
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Hazard Operability Analysis (HAZOP): A Quality Risk Management tool
N Vishal Gupta, Charan H Y.
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Sterilization of Porous Loads by Hphv Steam Sterilizer : A Review
R.G. Mukunda, N. Vishal Gupta
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Microwave assisted Synthesis and Characterization of Cu2+ doped ZnO Nano particles
P.Malarkodi, J.C. Kannan
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Ram Mohan S.R, N. Vishal Gupta
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Manu .C, N. Vishal Gupta
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Review on Recent developments and biological activities of 2, 4-thiazolidinediones
Ayyakannu Arumugam Napoleon
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Validated RP HPLC Method for the Determination of Related Substance of
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Evaluation of Antibacterial and Cytotoxic activity of Green Synthesized Cobalt
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International Editorial Board
Managing Executive Editors[Dev, A & R]
1
Editor-in-Chief: [from 17th Feb 2018]
Dr. Men Zhu, Ph.D.,
*Formulation Scientist, Verseon Corporation, 47071 Bayside Parkway
Fremont, CA 94538, USA.
47071 Bayside Parkway, Fremont, California 94538, USA
Tel: +1 (408) 769-9052
Email id: mzhu@verseon.com
Associate Editors
2
Juan Pablo Rodríguez Miranda
Distrital Francisco Jose de Caldas University. Faculty of Environment and
Natural Resources. Bogota D.C. Colombia.
[Official Representative of Journal in Latin America, Colombia and the
Caribbean]
Email id: jprm577@gmail.com
Telephone Mobile: +57 311 5319723
3
Dr. Rajendra B. Kakde,
Rashtrasant Tukadoji Maharaj Nagpur University, Campus, Nagpur 440033 ,
MS, India
Coordinator/Dean Faculty of Pharmaceutical Sciences
Academic Council Member, Director RUSA Centre
Former Senate Member, Director of Technology Park,
Coordinator for Patent Cell, Member of University Industry Interaction Cell,
Former Research Scientist Cadila, Lupin, Glenmark R& D.
Contact No: +919822711060
Email :
drkakde@yahoo.com
www.nagpuruniversity.org
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Dr. Ahmed A. Abd-Rabou
Researcher (PhD holder), Medical Biochemistry and Cancer Biology,
Medical Research Division, National Research Center (NRC), Egypt
(Permanent; 2008 up to date).
Post Doctor, Institut national de la sante et de la recherche medicale
(INSERM), Dijon (2016).
Fellow, Cancer Metabolism, Zewail City of Science and Technology
(ZCST), Egypt (2012 - 2016).
Fellow, Nano Drug Delivery, Albany College of Pharmacy and Health
Sciences (ACPHS), NY, USA (2013 - 2014).
Email id: ahmedchemia87@yahoo.com ,
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Postdoctoral researcher, Room 322, Pharmaceutical and biomedical
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Professeur de Enseignement Superieur Assistant Universite Sidi Mohamed
Ben Abdellah de Fes Institut National des Plantes Medicinales et
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Executive Editors[A & R]
1
Dr. Fazlul Huq,
Head Cancer Research Group, School of Medical Sciences, Faculty of
Medicine, Australia.
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Dr.Pavel Mokrejs,
Thomas Bata University,The Czech Republic, Europe.
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Dr. Adli Abdallah Hanna,
National Research Centre, Egypt.
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University of Mumbai,India.
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Arak University,Iran .
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University of Pitesti, Romania.
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University Negeri Yogyakarta, Indonesia.
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Centre Lab of Chem.Tech.,Poland.
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School of Chemical Sciences Universiti Sains Penang, Malaysia.
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Pasteur Institute of Iran.
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College of Health Sc.,PAAET, Kuwait.
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Hebei University, China.
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Zigzag University,Egypt
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University of Athens, Athens, Greece.
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IROST,Iran
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University PutraMalaysia, Malaysia.
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GSPS.IOP, M.S.India.
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Birla Insti.of Tech.,India.
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Prof. Dr. Sbirna Simona-Liana,
University of Craiova, Romania.
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Dr Carolyn Coulter,
University of Otago, New Zealand.
22
Dr. Konstantinos M. Kasiotis,
B P Institute, Laboratory of Pesticides Toxicology, Athens, 14561, Greece.
23
Dr. Arun HS Kumar,
University College Cork,Ireland.
Broccoli Flower Extract (Brassica oleracea L. var.italica
Plenck) Inhibits Photoaging by Increasing Type I
Procollagen Expression in Human Skin Fibroblast
Nelva K. Jusuf*
Department of Dermatovenereology Faculty of Medicine University of Sumatera Utara,
Indonesia
Abstract: Photoaging of the skin is caused by intrinsic process superimposed with degenerative changes due to environmental sources such as ultraviolet irradiation. Ultraviolet B (UVB) reduces type I procollagen level and increases matrix metalloproteinase-1 (MMP-1) level in human skin which plays a major role in the process of photoaging. Broccoli flower extract (BFE) is a crucciferae group of vegetables which has multiple antioxidants. It had been studied, acting as MMP-1 inhibitor agent both at mRNA and protein level on skin photoaging in vitro. BFE were investigated for their capacity to regulate type I procollagen expression at protein level in primary human fibroblast culture irradiated by UVB 50 mJ/cm2 and 100 mJ/cm2. Type I procollagen protein expression was quantified by Enzyme Immuno Assay. The result of studies showed pre-treatment with various concentration of BFE increase type I procollagen expression. There were significant differences of the mean value of type I procollagen expression based on irradiation dose (p<0.05) and BFE concentration (p<0.05). There was also interaction between irradiation dose and BFE concentration (p<0.05). There was positive correlation between BFE concentration with type I procollagen expression. Therefore BFE has been proved to increase type I procollagen expression at protein level on UVB irradiated human skin fibroblast. BFE has a potential effect as antiphotoaging agent in the near future.
Introduction
The skin aging process can be divided into intrinsic aging and extrinsic aging. Ultraviolet (UV) irradiation cause premature aging which is called as photoaging. It is caused by intrinsic processes superimposed with degenerative changes to solar radiation.1,2,3 UV irradiation disrupts the skin collagen matrix by stimulating collagen degradation by matrix metalloproteinase (MMP) and inhibiting procollagen production. The balance of MMP-1 and type I procollagen expression play an important role in photoaged skin. Alterations and defficiencies of collagen have been suggested to be a cause of the skin wrinkling observed in photoaged and naturally aged skin.4 Collagen degradation is mainly regulated by MMPs, while collagen synthesis is mediated by both transcriptional (gene/mRNA level) and post translational (protein level) processes.5 Broccoli (Brassica oleracea L.var.italica Plenck) is a cruciferous vegetables which has a great amount of antioxidant. It has been known that it consists of sulphoraphane, indole, beta carotene, quercetine, kaempferol, gluthatione, vitamin A, C, E, selenium.6,7 In dermatology it has been proved as skin antinflammatory in UV- induced erythema8 and antimutagenic on epidermolysis bullosa.9 In previous studies we found that broccoli flower extract(BFE) down regulated MMP-1 mRNA expression and MMP-1 protein expression on UVB irradiated human fibroblast cell culture.10
Nelva K. Jusuf / International Journal of PharmTech Research, 2016,9(3),pp 114-118. 115
Experimental
Material and methods
Plant material
Broccoli flowers were collected from broccoli field at Berastagi, North Sumatera, Indonesia. After processed to a standardized simplicia it was extracted with ethanol 96% in the laboratory of Faculty of Pharmacy University of Sumatera Utara, Indonesia.
Cell culture
The normal human fibroblast cells were aseptically isolated from preputium circumcised skin of healthy volunteer age 11-12 years old. After the epidermis and dermis were separated mechanically, the dermis was minced and attached on the surface of tissue culture flask. The cells were grown in Dulbecco’s modified eagle’s medium (DMEM) supplemented with 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin (GIBCO,Grand Island, NY USA). After 3 passages the fibroblast were used for the experiment.
Ultraviolet irradiation
The UV light source originated from a Philips TL 20 W(12 RS fluorescent sun lamp with an emission spectrum of 285-350 nm (peak at 310-315 nm). The cells were then exposed to 50 and 100 mJ/cm2 dose of UVB light.
Treatment with BFE
BFE was dissolved in DMEM. The BFE concentration that use for treatment comprise of 25, 50 and 100 mg/ml. For treatment, the cells were maintained in culture media without FBS overnight, followed by treatment with BFE for 24 hours. The cells were rinsed twice with phosphate buffer saline (PBS) and UVB irradiation exposure were performed under a thin layer of PBS. Immediately after irradiation, the cells were incubated in serum-free fresh culture media containing BFE.
Enzyme Immuno Assay (EIA)
The supernatant from the cell culture was collected and the procollagen type I protein expression was quantified by procollagen type I C peptide EIA kit (Takara Biomedical Inc. Japan) at 450 nm using a microplate reader.
Statistical Analysis
Data were expressed as the mean value and with Fisher’s least significant difference (LSD) were analyzed by analysis of variance (Anova) 2 ways, continued by multiple comparison test. Statistical significance was set a prior at p<0.05. The correlation between BFE concentration and type I procollagen protein expression were analyzed by Spearman’s correlation test.
Result
Effect of BFE on type I procollagen protein expression
Figure 1. Effect of BFE on type I procollagen protein expression
Table 1. Mean value of type I procollagen protein expression between every group based on irradiation dose and BFE concentration.
Correlation between BFE concentration with type I procollagen protein expression
By Spearman’s correlation test we found there were significant positive correlation between BFE concentration with type I procollagen protein expression at 50 mJ/cm2 UVB irradiation dose (r=0.975, p<0.01) and at 100 mJ/cm2 UVB irradiation dose (r=0.973, p< 0.01) (Table 2).
Table 2.Correlation between BFE concentration with type I procollagen protein expression.
n r p
Procollagen type I protein (50) 12 0,975 0,0001 Extract concentration
Procollagen type I protein (100) 12 0,973 0,0001
Spearman’s correlation test; Correlation is significant at the 0,01 level (2-tailed); 50: UVB irradiation dose 50 mJ cm-2;100: UVB irradiation dose 100 mJ cm-2; r: correlation coefficient.
Discussion
Nelva K. Jusuf / International Journal of PharmTech Research, 2016,9(3),pp 114-118. 117
further processed to be a collagen fiber in the extracellular matrix.13 Photoaging is a process involving alteration of type I collagen which is the major component of the dermis. Among collagen, type I is the most abundant and comprises between 85% of the total collagen in the skin. Type I collagen degradation is the major cause of wrinke formation.14 The use of antioxidant is an effective approach to prevent symptoms related to photo-induced aging of the skin.Strategies to prevent or at least minimize ROS-photo-induced photoaging necessarily include protection against UV irradiation and antioxidant homeostasis.15
Broccoli flower extract (Brassica oleracea var.italica Plenck) is an antioxidant plant which has been studied in dermatology. Sulphorapane, the most potent antioxidant in broccoli, has antiinflammatoric effect which is shown by preventing erythema from sunlight.8 In our previous study, we reported that BFE decreased the level of MMP-1 expression both at mRNA and protein level in the process of photoaging induced by UVB irradiated in vitro.10Therefore on the other hand we investigated wether BFE also may increase the level of type I procollagen expression. Our results showed that BFE has enhancer activity on type I procollagen protein expression in UVB irradiated human skin fibroblast. We also found there were interaction between UVB irradiation dose and BFE concentration to the expression of type I procollagen. This result indicated that both factors together influence the mean value of type I procollagen expression.We also found there were positive correlation between BFE concentration with type I procollagen protein expression, thereby the increasing of BFE concentration leads the type I procollagen expression increased. These results demonstrates the inducer effect of BFE on type I procollagen expression via protein assay in skin photoaging process in vitro. Therefore it is suggested that BFE should be viewed as a potential therapeutic agent for preventing and treating skin photoaging based on decrease the MMP-1 expression and increase procollagen type I expression.
Conclusion
BFE had inhibitory effect on skin photoaging induced by UVB in vitro by increasing type I procollagen expression at protein level in human skin fibroblast culture. It is suggested further studies on BFE involving human subjects should be carried out in the near future.
Acknowledgement
Author thanks to Prof.Dr.dr Hardyanto Soebono, Sp.KK(K), Prof.Drs. Sumadio Hadisahputra, Apt., Ph.D, dr. Adang Bachtiar, MPH, D.Sc, Dr.dr. Imam Budi Putra, MHA, SpKK for support and cooperation. The author have no funding or support to report. No conflict of interest.
References
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2. Jenkins, G. Molecular mechanism of skin ageing: Mech. Ageing Dev. 2000 vol. 123, pp. 801-10. 3. Yaar, M & Gilchrest, BA. Aging of skin. in: Wollf K, Goldsmith LA, Katz SI, Gilchrest BA, Paller AS,
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6. Ipteknet Sentra Informasi IPTEK . Tanaman obat Indonesia: Brokoli. 2005 viewed 11 Maret 2010. <http://www.iptek.net.id/ind/pd_tanobat/view.php?mnu=2&id=20?>.
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