Background: (TTD) is a zinc-specific popularly used to treat chronic alcoholism by targeting zinc-dependent alcohol enzyme. Interestingly, enzyme (ACE), neutral (NEP) and N (APN) implicated in hypertension are zinc-dependent enzymes.
Objective: This study aimed to evaluate the inhibitory potential of TTD against ACE, NEP, and APN in vitro. Further, the anti-hypertensive and efficacy of TTD in were evaluated using N (Ï‰)-nitro-L-arginine-methyl ester (L-NAME)-induced hypertensive rat model.
Method: Dose-dependent inhibition of ACE, NEP, and APN by TTD was evaluated in vitro. Further, five groups consisting of 6 rats in each group were used for animal studies. Group one served as control, and the second group received L-NAME alone (40mg/kg/day). The third group was TTD control. Fourth and fifth group received L-NAME and simultaneously treated with TTD (10mg/kg/day) and (10mg/kg/day) respectively. Systolic blood pressure, heart and kidney weight; angiotensin, atrial natriuretic peptide urea, creatinine, aspartate transaminase, alanine transaminase, and nitrite levels were measured in serum of all group of rats. Fibrosis in heart and kidney tissues were evaluated by staining collagen with red.
Results: TTD effectively inhibited the activities of ACE, NEP, and APN in a dose-dependent manner with IC50 of 10.15 ÂµM, 22.35 ÂµM and 3.032 ÂµM respectively. Further, administration of the TTD effectively decreased systolic blood pressure (140Â±8 mmHg) when compared to L-NAME group (190Â±10 mmHg), improved hypertension markers (Ang II and ANP), kidney and liver markers. TTD ameliorated heart and kidney hypertrophy and partially prevented fibrosis.
Conclusion: These findings provide evidence that TTD affords efficient anti-hypertensive effect with moderate end-organ protection possibly by cumulative inhibition of the aforementioned enzymes.
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Introduction: Dendritic cell therapy is a promising therapeutic therapy for cancer. Various methods have been developed to culture and expand dendritic cells in vitro. However, most methods have used fetal bovine serum (FBS)-supplemented media to induce and expand monocytes; cells prepared by these methods cannot be used in the clinic. Therefore, this study aims to develop new methods to produce dendritic cells (DCs) from monocytes using serum-free medium.
Methods: mononuclear cells (MNCs) were isolated from human umbilical cord blood by gradient centrifugation. They were then induced into DCs using 3 kinds of inducing media: M1 (medium supplemented with FBS), M2 (medium supplemented with human serum (HS) from umbilical cord blood), and M3 (medium supplemented with platelet-rich plasma (PRP) from umbilical cord blood) at 10% supplement. The MNCs were induced to immature DCs (iDCs) by granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin (IL)-4, and matured by tumor necrosis factor (TNF)-alpha. The phenotype of the DCs were evaluated by flow cytometry, immunohistochemistry, and in vitro phagocytic assay.
Results: The results showed that all cells in the groups exhibited the shape of dendritic cells.Immunohistochemistry analysis showed that the mature cultured cells in the 3 kinds of media (i.e. supplemented with either FBS, HS, or PRP) were all CD86+HLA-DR+CD14-, representative of mature DC (mDC) phenotype. DCs cultured in HS and PRP media also exhibited FITC-Dextran phagocytosis and showed IL-12 gene expression similar to those DCs cultured in FBS medium.
Conclusion: The results of the present study suggest that HS or PRP can replace FBS to produce DCs in vitro for clinical use.
Biology is one the most exciting disciplines. Biology is a science subject that can effect on all fields of the life. Day by day, biology participates to a lot of fields included agriculture, medicine, pharmacology, biotechnology, environmental protection, and conservation. It covers from these smallest molecules to the ecosystems.