Jan 2020 Faizul Jaafar   1 ,  Lina Wati Durani   1 ,  Suzana Makpol   2   1. Department of Biochemistry, Faculty of Medicine, Level 17, Preclinical Building, Universiti Kebangsaan Malaysia Medical Centre, Jalan Yaakob Latif, Bandar Tun Razak, Cheras, 56000, Kuala Lumpur, Malaysia.   2. Department of Biochemistry, Faculty of Medicine, Level 17, Preclinical Building, Universiti Kebangsaan Malaysia Medical Centre, Jalan Yaakob Latif, Bandar Tun Razak, Cheras, 56000, Kuala Lumpur, Malaysia. suzanamakpol@ppukm.ukm.edu.my . Abstract Human diploid fibroblasts (HDFs) cultured in vitro have limited capacity to proliferate after population doubling is repeated several times, and they enter into a state known as replicative senescence or cellular senescence. This study aimed to investigate the effect of Chlorella vulgaris on the replicative senescence of HDFs by determining the expression of senescence-associated genes. Young and senescent HDFs were divided into untreated control and C. vulgaris-treated groups. A senescence-associated gene transcription analysis was carried out with qRT-PCR. Treatment of young HDFs with C. vulgaris reduced the expression of SOD1, CAT and CCS (p < 0.05). In addition, the expression of the SOD2 gene was increased with C. vulgaris treatment in young, pre-senescent and senescent HDFs (p < 0.05). Treatment of senescent HDFs with C. vulgaris resulted in the downregulation of TP53 gene expression. The expression of the CDKN2A gene was significantly decreased upon C. vulgaris treatment in young and senescent HDFs. C. vulgaris treatment was also found to significantly upregulate the expression of the MAPK14 gene in pre-senescent HDFs. In addition, the expression of MAPK14 was significantly upregulated compared to that in the untreated senescent HDFs (p < 0.05). In summary, the expression of senescence-associated genes related to antioxidants and the insulin/insulin-like growth factor-1 signalling, DNA damage-associated signalling, cell differentiation and cell proliferation pathways was modulated by C. vulgaris during replicative senescence of human diploid fibroblasts. Keywords:  Chlorella vulgaris; Fibroblasts; Replicative senescence; Senescence-associated genes.

August 2020 Tomohiro Bito   1 ,  Eri Okumura   2 ,  Masaki Fujishima   2 ,  Fumio Watanabe   1   1. Department of Agricultural, Life and Environmental Sciences, Faculty of Agriculture, Tottori University, Tottori 680-8553, Japan. 2. Sun Chlorella Corporation, Kyoto 600-8177, Japan. Abstract Chlorella  is a green unicellular alga that is commercially produced and distributed worldwide as a dietary supplement.  Chlorella  products contain numerous nutrients and vitamins, including D and B12, that are absent in plant-derived food sources.  Chlorella  contains larger amounts of folate and iron than other plant-derived foods.  Chlorella  supplementation to mammals, including humans, has been reported to exhibit various pharmacological activities, including immunomodulatory, antioxidant, antidiabetic, antihypertensive, and antihyperlipidemic activities. Meta-analysis on the effects of  Chlorella  supplementation on cardiovascular risk factors have suggested that it improves total cholesterol levels, low-density lipoprotein cholesterol levels, systolic blood pressure, diastolic blood pressure, and fasting blood glucose levels but not triglycerides and high-density lipoprotein cholesterol levels. These beneficial effects of  Chlorella  might be due to synergism between multiple nutrient and antioxidant compounds. However, information regarding the bioactive compounds in  Chlorella  is limited. Keywords:  Chlorella; antioxidants; dietary fibers; dietary supplements; folate; lutein; vitamin B12; vitamin D2.

Hyo-Jin An 1, Hong-Kun Rim, Hyun-Ja Jeong, Seung-Heon Hong, Jae-Young Um, Hyung-Min Abstract The objective of this study was to investigate the effects of hot water extracts of Chlorella vulgaris (CVE) on a deteriorated immune function through utilization of a protein-energy malnutrition (PEM) diet. Unicellular algae, C. vulgaris, were used as biological response modifier. PEM is associated with decreased host immune defense. Male C57BL/6J mice, initially four weeks old, were fed for 8 days with standard diet or a PEM diet. Mice in the PEM diet group were orally administered 0.1 g/kg and 0.15 g/kg of CVE for the following week. Nutritional parameters such as the total protein, albumin, glucose, and interferon γ (IFN-γ) were increased in blood serum of the CVE-treated group compared with the non-treated group. The mononuclear cell numbers from spleen, superficial, and mesenteric lymph node were reduced in mice fed with PEM diet, but numbers from the spleen and superficial lymph node were increased by the CVE (0.1 and 0.15 g/kg) treatment. We also investigated the effect of CVE on the production of cytokines in human T-cell line, MOLT-4 cells, and primary cultured splenocytes. The CVE treatment significantly increased the production of both interleukin (IL)-2 and IL-4 compared with the media control, but did not affect the production of IFN-γ. These results suggest that CVE may be useful in improving the immune function.

July 2010   Na-Hyung Kim   1 ,  Kyu-Yeob Kim ,  Hyun-Ja Jeong ,  Hyung-Min Kim ,  Seung-Heon Hong ,  Jae-Young Um Department of Pharmacology, College of Oriental Medicine, Institute of Oriental Medicine, Kyung Hee University, Seoul, Republic of Korea. Abstract Background:  Chlorella vulgaris is a unicellular and microscopic algae that is currently used in a variety of forms of tablets, capsules and liquid as a biological response modifier. The aim of this study was to investigate the effects of hydrolyzed Chlorella vulgaris by malted barley for its potential reduction of the immobility time in ICR mice and on the cytokine regulation in human T cell line, Molt-4. Results:  After a forced swimming test, the changes in aspects of blood biochemical parameters due to the administration of hydrolyzed Chlorella vulgaris by malted barley were examined. The effect of hydrolyzed Chlorella vulgaris by the malted barley-treated group for 14 days on the immobility time was significantly reduced in comparison with that of the control group (P < 0.01). The plasma level of blood urea nitrogen was significantly decreased in hydrolyzed Chlorella vulgaris by malted barley-treated group compared with the control group (P < 0.05). In addition, hydrolyzed Chlorella vulgaris by malted barley increased interferon-gamma and interlukin-2 levels in Molt-4 cells. Conclusion:  These results indicate that hydrolyzed Chlorella vulgaris by malted barley is useful for immune function improvements, enhanced physical stamina, and as a candidate for an anti-fatigue or antidepressant agent.

2006   Hyo-Jin An   1 ,  Hyun-Myung Choi ,  Hyeung-Suk Park ,  Jae-Gab Han ,  Eun-Hee Lee ,  Young-Sig Park ,  Jae-Young Um ,  Seung-Heon Hong ,  Hyung-Min Kim   College of Oriental Medicine, Institute of Oriental Medicine, Kyung Hee University, Dongdaemun-Gu, Seoul, Korea.   Abstract Background/aims:  A unicellular algae, Chlorella vulgaris, was used as a biological response modifier. Although hot water extracts of C. vulgaris (CVE) are thought to augment immune responses, the effect of CVE on fatigue and physical stamina has not been studied. Methods:  In the present study, we investigated the effect of CVE on forced swimming test and blood biochemical parameters related to fatigue, blood urea nitrogen (BUN), creatine kinase (CK), lactic dehydrogenase (LDH), glucose (Glc), and total protein (TP). CVE (0.05-0.15 g/kg/day) was orally administered to mice. Results:  After 7 days, the immobility time was decreased in the 0.1- and 0.15-g/kg CVE-treated groups (179 +/- 8.3 and 175 +/- 2.1 s) in comparison with the control group (223 +/- 5.4 s). In addition, the contents of BUN, CK, and LDH in the blood serum were decreased in the CVE-fed group. However, they had no effect on the elevation of Glc and TP level. Conclusions:  The results predict a potential benefit of CVE for enhancing immune function and improving physical stamina. Copyright 2006 S. Karger AG, Basel.

May 1994  T Hasegawa   1 ,  M Okuda ,  K Nomoto ,  Y Yoshikai Laboratory of Germfree Life, Nagoya University School of Medicine, Japan. Laboratory of Germfree Life, Nagoya University School of Medicine, Japan. Abstract Oral administration of a hot water extract of Chlorella vulgaris(CVE)(20mg/mouse, 10 consecutive days) augmented the resistance against an i.p. infection with Listeria monocytogenes in mice. The numbers of bacteria in a CVE-administered group were significantly lower in the peritoneal cavity or spleen than those in a control group. FCM analysis revealed that gamma delta +Thy1.2+ cells in the nonadherent PEC from CVE-administered mice increased more prominently in number at the early stage on day 3 or on day 5 after infection as compared with those in control mice. The increment of gamma delta +Thy1.2+ T cells was also evident in spleen in CVE-administered mice at this stage after infection. The proportion of TCR alpha beta +Thy1.2+ T cells in the nonadherent PEC of a control group increased from 13% on day 0 to 49% at the late stage on day 10 after infection, whereas the proportion of TCR alpha beta +Thy1.2+ T cells in the nonadherent PEC in CVE-administered mice increased to 64% on this stage after infection in association with augmentation of DTH response to Listeria. These results suggest that CVE-administration effectively augment cell-mediated immunity against Listeria through the increment of gamma delta + T cells in the early phase and the increment of alpha beta + T cells in the late phase after listerial infection.

January 1997 T Hasegawa   1 ,  Y Kimura ,  K Hiromatsu ,  N Kobayashi ,  A Yamada ,  M Makino ,  M Okuda ,  T Sano ,  K Nomoto ,  Y Yoshikai Research Laboratories, Chlorella industries Co., Ltd., Fukuoka, Japan.   Abstract We have previously reported that oral administration of hot water extract of Chlorella vulgaris (CVE) enhances resistance to Listeria monocytogenes through augmentation of Listeria-specific cell-mediated immunity in normal mice and mice with murine acquired immunodeficiency syndrome (MAIDS) caused by murine leukemia virus (MuLV) LP-BM5. To elucidate the mechanisms whereby CVE augments the cell-mediated immunity, we examined the expression patterns of mRNA for cytokines in normal and MAIDS mice given CVE orally after L. monocytogenes infection. The expression levels of IL-1 alpha, IL-12, GM-CSF, MIP and TNF alpha genes were significantly augmented in the peritoneal adherent cells by oral administration of CVE for 2 weeks before Listeria infection. The expression levels of gamma IFN and IL-12 mRNA were significantly higher in the spleen after Listeria infection in CVE-treated mice than in normal mice, while the expression of IL-10 mRNA in the spleen was decreased by CVE administration. In MAIDS mice, oral administration of CVE also augmented the expression of gamma IFN and IL-12 mRNA in the spleen after Listeria infection, while it rather reduced the expression of IL-10 mRNA. These results suggest that CVE may preferentially augment THI responses against Listeria via activation of macrophages to produce IL-12 and enhance host defence against Listeria infection both in normal and MAIDS mice.

May 1999 T Hasegawa   1 ,  K Ito ,  S Ueno ,  S Kumamoto ,  Y Ando ,  A Yamada ,  K Nomoto ,  Y Yasunobu Research Laboratories, Chlorella Industries Co., Ltd, Chikugo City, Fukuoka, Japan. hasegawa@chlorella.co.jp Abstract Hot water extract of Chlorella vulgaris (CVE) is a biological response modifier (BRM) which enhances resistance to Listeria monocytogenes through augmentation of helper T cell type 1 (Thl) responses producing gamma-interferon (gammaIFN). We show here that oral administration of CVE in mice suppressed the production of immunoglobulin (Ig)E against casein antigen accompanied by increased gammaIFN and IL-12 mRNA expression. Oral administration of CVE enhanced Thl response to casein in the spleen of casein immunized mice. CVE may be useful for prevention of allergic diseases with a predominant Th2 response.

June 2016   Byoung-Ki An   1 ,  Kwan-Eung Kim   2 ,  Jin-Young Jeon   2 ,  Kyung Woo Lee     1. Laboratory of Poultry Nutrition, Department of Animal Science and Technology, College of Animal Bioscience and Technology, KonKuk University, 120 Neungdong-ro, Gwangjin-gu, Seoul, 143-701 South Korea. 2. Department of Health Food R&D, Daesang Corp., Icheon, 467-813 Korea. Abstract This experiment was carried out to investigate the effects of dried chlorella powder (Chlorella vulgaris; DCP) and chlorella growth factor (CGF) on growth performance, serum characteristics, meat qualities and humoral immune responses in broiler chicks. A total of 1050 day-old Ross male broiler chicks were randomly divided into 35 pens (30 chicks/pen) and subjected to one of seven dietary treatments. A non-medicated corn-soybean meal base diet was considered as negative control (NC) and added with either antibiotic (PC), three levels of DCP (NC diets added with 0.05, 0.15 or 0.5 % DCP) or two levels of CGF (NC diets added with 0.05 or 0.15 % CGF). The final body weight and daily weight gain in PC and groups fed diets with 0.15 or 0.5 % DCP were heavier (p < 0.001) than those of NC and CGF-treated groups. Serum total lipid concentrations were lower (p = 0.001) in groups fed diets with 0.5 % DCP and 0.05 or 0.15 % CGF compared with PC group. The levels of serum IgG (p = 0.050) and IgM (p = 0.010) were elevated in chicks fed diets with DCP and CGF compared with the PC or NC group. Meat qualities such as cooking loss, meat color, and pH, of edible meats were not altered by dietary treatments. Collectively, these results indicate that dietary DCP, but not CGF, exerted growth-promoting effect, and both DCP and CGF affected humoral immune response in broiler chicks.  Keywords:  Chlorella growth factor; Chlorella vulgaris; Growth performance; Humoral immune response.

August 2002 Mary L S Queiroz   1 ,  Claudia Bincoletto ,  Marize C Valadares ,  Denise C M Dantas ,  Leonilda M B Santos Departamento de Farmacologia/Hemocentro, Faculdade de Ciências Médicas, Universidade Estadual de Campinas.   Abstract In this study, we have investigated the effects of the unicellular-green-algae Chlorella vulgaris on the production of INF-gamma, IL-2, IL-4 and IL-10 in normal and Listeria monocytogenes infected mice. Our results demonstrated that in normal/non infected mice, CVE administration produced no effects in the levels of all cytokines studied. However, Listeria monocytogenes infection enhanced the production of INF-gamma and IL-2 at 48 and 72 h after the bacteria inoculation. Interestingly, the treatment with five consecutive doses of 50 mg/Kg/day of Chlorella vulgaris given previously to infection, led to further increases in INF-gamma and IL-2 levels at 48 and 72 h in relation to the presence of infection alone. No changes in IL-4 and IL-10 production were observed in Listeria monocytogenes and CVE treated/infected mice. These results are in accordance with the literature, which shows that CVE is a biological response modifier that enhances resistance to Listeria monocytogenes through augmentation of IL-2 and IFN-gamma.

Hasegawa T, Yoshikai Y, Okuda M, Nomoto K. Int J Immunopharmacol 1990 12:8 883-91

Konishi F, Tanaka K, Himeno K, Taniguchi K, Nomoto K. Cancer Immunol Immunother 1985 19:2 73-8 Abstract When a hot water extract of Chlorella  vulgaris (CE) was injected into the peritoneal cavity of BALB/c mice inoculated with syngeneic Meth-A tumor cells, the survival times were strikingly prolonged. Furthermore, peritoneal exudate cells (PEC) rich in polymorphonuclear cells (PMN) obtained from normal mice 24 h after CE injection exhibited an antitumor effect in a Winn-type assay using normal recipients. Such an activity of PEC remained almost intact after T cell or macrophage depletion. However, such PEC did not express an antitumor effect in a Winn-type assay using irradiated recipients. It was suggested that CE-induced PEC, presumably PMN, expressed an antitumor effect in cooperation with a host- or recipient-derived element(s) sensitive to irradiation. The antitumor mechanism of CE may be different from that of OK-432, one of the biological response modifiers