Diana Lyrawati
Laboratorium Farmasi Fakultas Kedokteran Universitas Brawijaya Malang

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Journal : Journal of Tropical Life Science : International Journal of Theoretical, Experimental, and Applied Life Sciences

Date Fruit Extract (Phoenix dactylifera, Ajwa) Modulates NK Cells and TNF- Alpha in DMBA-Induced Mammary Cancer Sprague-Dawley Rats Elhemeidy, Rehab Mohammed Mustofa; Lyrawati, Diana; Widjajanto, Edi
Journal of Tropical Life Science Vol 8, No 3 (2018)
Publisher : Journal of Tropical Life Science

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.11594/jtls.08.03.04

Abstract

Cellular immune system is one of anti-tumor target development. NK cells, one of major components of antitumor innate immune responses, are frequently decreased functionally during breast cancer progression, whereas TNF-alpha changes from anti-tumor into pro-tumorigenesis and its level increases with advancing cancer. In this study, we investigated the immunomodulatory effects of Ajwa date fruit extract to determine whether it stops the progression of breast cancer. Using rats induced with single dose 20 mg DMBA subcutaneously, we examined whether administration of Ajwa date fruit (Phoenix dactylifera) extract (at 400, 800 mg/kg body weight/d) post-DMBA induction for 30 days modulate NK cells, TNF-alpha, and development of breast cancer. ELISA, FACS, immunohistochemistry, and histologic observation were employed. Compared to positive control group (DMBA-induced only), we showed that the Ajwa date fruit extract normalize the level of circulatory CD161 NK cells and breast tissue TNF-alpha, cell size and proliferation, and improve overall survival rates. Ajwa date fruit extract may be used to modulate NK cells and TNF-alpha against progression of breast cancer. Further studies are required to develop date fruit extract as anti-cancer in human, whether as single or adjunct therapy to standard management of breast-cancer.
Agarose Coated Culture Plate in Tumorsphere Culture of Cervical Cancer Cell Line HeLa: an Alternative to Non Adhesive Culture Plate Juniartha, Putu; Indra, Muhammad Rasjad; Sujuti, Hidayat; Lyrawati, Diana; Nurseta, Tatit
Journal of Tropical Life Science Vol 6, No 3 (2016)
Publisher : Journal of Tropical Life Science

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.11594/jtls.06.03.11

Abstract

Cervical cancer recurs in 90% cases and linked to cancer stem cells that able to self-renew and responsible for recurrence, metastasis, and mortality of cancer. Isolation and identification of cancer stem cells using serum-free medium needs expensive growth factors and consume time. This study try to grow tumor sphere using culture plate coated with 1% agarose as an efficient and economical alternative to non-adhesive culture plate. HeLa cell line was grew in culture plate coated with 1% agarose and non-adhesive culture plate using similar medium and culture condition. Tumor spheres morphology was observed and the colonies were counted in 7 days followed by single cell assay. Tumor spheres then counted for CD133+, CD34+, and Sox2 expression using flowcytometry. Culture plate coated with 1% agarose can be used as an economic and efficient alternative to culture tumor sphere. Using culture plate coated with 1% agarose, the tumor spheres formed in 7 days with similar morphology to non-adhesive culture plate. Tumorsphere had three dimensional – sphere shape that tightly attached, colonized, and overlapped. The tumor sphere colony counts of two plates were statistically have no significant difference (p=0,667). Single cell assay of a tumor sphere shows that it can grow new tumor spheres with similar morphology. The tumor sphere from culture plate coated with 1% agarose express CD133+ and CD34+ as much as 8.78% ± 2.14 and Sox2 as much as 35.30% ± 23.82 whereas tumor sphere from non-adhesive culture plate express CD133+ and CD34+ as much as 62.36% ± 1.06 and Sox2 as much as 98.86% ± 0.56 (p = 0000).
Role of Antibody Anti-AGE in the Expression of Nephrin and RAGE on Primary Glomerulus Cell Exposed with AGE Salam, Rudy; Lyrawati, Diana; Samsu, Nur
Journal of Tropical Life Science Vol 7, No 2 (2017)
Publisher : Journal of Tropical Life Science

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.11594/jtls.07.02.03

Abstract

Upregulation of nephrin expression occurs at the early stage of nephropathy and decrease in the period 6 months on hyperglycaemic condition. Nephrin is associated with the initial stage of the loss of the permeability barrier in diabetic nephropathy. Interaction AGE-RAGE increases angiotensin II on Renin Angiotensin-Aldosterone System (RAAS) and activation of protein kinase c (PKC) which induce alterations in nephrin mRNA expression. Alterations of nephrin expression induces transformation of slit membrane structure and the permeability changes at the glomerular filtration barrier. Anti-AGE vaccination once may cause the changes of nephrin and RAGE expression and can prevent progression of diabetic nephropathy. This study used primary glomerulus cell culture obtained from renal of Wistar mice aged 3 months, weighting 200-300 grams that consist of negative control group that exposed to BSA 100 µg/ml, positive control group that exposed to AGE-BSA 100 µg/ml, treatment group 1 that exposed to polyclonal anti-AGE 5 µg/ml and AGE-BSA 100 µg/ml and treatment group 2 that exposed to monoclonal antibody anti-CML 5 µg/ml and AGE-BSA 100 µg/ml. Paired t-test with a 0.05 level of confidence results showed that there were significant differences in level of RAGE expression among experimental groups with control groups. Administration of polyclonal antibody decreased RAGE expression among negative control (p=0.188). but not in positive control (p=0.000). In contrast to monoclonal anti-AGE antibody, RAGE expression did not differ significantly compared to negative control but significant than positive control. Administration of monoclonal anti-AGE antibody inhibited increasing of nephrin expression compared to negative and positive control (p=0.73; 0.125). In conclusion, this study suggested that administration of polyclonal and monoclonal anti-AGE antibody could inhibit increasing of RAGE and nephrin expression in glomerulus primary culture that exposed to AGE which is expected to prevent the progression of diabetic nephropaty.
Advanced Glycation End Products (AGEs) Antibody Protects Against AGEs-induced Apoptosis and NF-ĸB p65 Subunit Overexpression in Rat Glomerular Culture Adianingsih, Oktavia Rahayu; Lyrawati, Diana; Samsu, Nur
Journal of Tropical Life Science Vol 6, No 3 (2016)
Publisher : Journal of Tropical Life Science

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.11594/jtls.06.03.08

Abstract

Advanced glycation end products (AGEs) have been thought to be a major cause of diabetic nephropathy (DN). The mechanisms underlying the involvement of AGEs antibody in diabetic nephropathy are not fully understood. The present study was designed to investigate the protective effect of AGEs antibody on AGEs-induced glomerular damage. Isolated glomeruli were pre-incubated either with 10 µg/mL polyclonal anti-AGEs antibody (AGE-pAb) or monoclonal anti-Nɜ -carboxymethyl-lysine antibody (CML-mAb) as a model of AGEs antibody to block interaction of AGEs with receptor for AGEs (RAGE) and incubated afterwards either with 100 µg/mL bovine serum albumin (BSA) or AGE-modified bovine serum albumin (AGE-BSA) for 48 h. Annexin V/nephrin doublestaining was performed to determine apoptosis. Using immunofluorescence, we found that administration of AGE-BSA not only significantly increased glomerular cells apoptosis and nuclear factor kappa B (NF-ĸB) p65 expression, but also reduced expression of nephrin, an important structural and signal molecule of podocytes slit diaphragm. Blocking the interaction of AGE-RAGE with AGEs antibody significantly protected glomerular cells from AGEs-induced apoptosis and NF-ĸB p65 overexpression. We found that AGE-pAb conferred superior protective effect compared with CmL-mAb for the same reduction in apoptosis and NF-ĸB p65 expression. In sharp contrast, CmL-mAb led to preserve expression of podocytes nephrin better than AGE-pAb. These results demonstrate that the antibody against AGEs may be beneficial for preventing the glomerular damage in DN.