Development and analytical validation of an enzyme-linked immunosorbent assay for the measurement of feline tumor necrosis factor α in serum

Abstract: Tumor Necrosis Factor- α (TNF-α) is a systemic pro-inflammatory cytokine which is responsible to a trauma, injury, or inflammation in human body. Physical activity can physiologically affect the human body, inter alia the immune system. This was an experimental study with a post test design. This study aimed to determine the profile of TNF-α after doing gymnastics among the elderly at nursing home Bethania Lembean. Samples included 30 peoples who met the inclusion criteria, i.e. over 64 years old, followed the gymnastics for 5 weeks regularly, approbated as respondents, healthy, and signed the informed consent. The TNF-α concentration measurements were done by using enzyme-linked immunosorbent assay (ELISA) Quantikine ®Human TNF-α. The results showed that the average conventration of TNF-α after elderly gymnastics was 70.54 pg/ml, with the highest value of 88.90 pg/mL and the lowest value of 12.54 pg/mL. For all respondents, TNF-α was within normal limit and ranged between 10-100 pg / mL. Conclusion: In this study, TNF-α concentrations after elderly gymnastics for 5 weeks were within normal limitsKeywords: TNF-α, elderly gymnasticsAbstrak: Tumor Necrosis Factor-α (TNF-α) merupakan suatu sitokin sistemik pro-inflamasi, yang berespons terhadap suatu cedera trauma atau peradangan yang terjadi dalam tubuh manusia. Kegiatan fisik dapat berdampak cukup besar pada tubuh manusia, dimana dapat merubah sistem imun tubuh secara fisiologis. Penelitian ini bersifat eksperimental dengan rancangan post test design dan bertujuan untuk mengetahui profil TNF-α sesudah senam lansia di Panti Wredha Bethania Lembean. Sampel penelitian berjumlah 30 orang yang memenuhi kriteria inklusi yaitu berumur di atas 64 tahun, teratur mengikuti senam lansia selama 5 minggu, bersedia menjadi responden, sehat saat di periksa, dan menandatangani informed consent. Pengukuran konsentrasi TNF-α di lakukan dengan menggunakan enzyme-linked immunosorbent assay (ELISA) Quantikine ®Human TNF-α. Hasil penelitian memperlihatkan bahwa rerata kadar TNF-α sesudah senam lansia 70,54 pg/ml, dengan nilai tertinggi 88,90 pg/mL dan nilai terendah 12,54 pg/mL. Berdasarkan hasil yang di dapat, semua kadar TNF-α masih dalam batas normal dengan nilai 10-100 pg/mL. Simpulan: TNF-α sesudah senam lansia selama 5 minggu masih dalam batas normal.Kata kunci:TNF-α, senam lansia

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Induction of matrix metalloproteinase-1 by tumor necrosis factor-α is mediated by interleukin-6 in cultured fibroblasts of keratoconus

Experimental Biology and Medicine ◽

10.1177/1535370216650940 ◽

2016 ◽

Vol 241 (18) ◽

pp. 2033-2041 ◽

Cited By ~ 7

Author(s):

Genlai Du ◽

Chengxing Liu ◽

Xiaona Li ◽

Weiyi Chen ◽

Rui He ◽

...

Keyword(s):

Tumor Necrosis Factor ◽

Matrix Metalloproteinase ◽

Interleukin 6 ◽

Tumor Necrosis ◽

Tumor Necrosis Factor Α ◽

Enzyme Linked Immunosorbent Assay ◽

Tnf Α ◽

Factor Α ◽

Inflammatory Molecules ◽

Necrosis Factor

Inflammatory molecules and matrix metalloproteinase (MMPs) have been found over-expressed in the tear film of patients with keratoconus. However, the mechanistic link between inflammatory molecules and MMPs in the pathogenesis of keratoconus remains still elusive. Therefore, we investigated the effect of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) on MMP-1 expression and used IL-6 antibody (IL-6 Ab) to examine the role of IL-6 on TNF-α mediated regulation of MMP-1 in fibroblasts of normal cornea and keratoconus. Real-time polymerase chain reaction, Enzyme-linked immunosorbent assay, and Western blot data demonstrated that MMP-1 and IL-6 were expressed in fibroblasts of normal cornea and keratoconus. Levels of MMP-1 and IL-6 were significantly higher in keratoconus than normal cornea. TNF-α treatment led to a significant increase in IL-6 levels. IL-6 treatment induced MMP-1 synthesis in normal cornea and keratoconus. TNF-α increased MMP-1 expression in a dose- and time-dependent manner and this response was completely inhibited by the IL-6 Ab. In conclusion, these results indicate that fibroblasts of keratoconus shows increased levels of IL-6 and MMP-1 gene and protein expression and IL-6 mediates the TNF-α-induced MMP-1 expression.

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