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Title:	Immunomodulatory effects of the extract and chemical constituents of Tinospora crispa (L) Hook. F. & thomson on various components of the innate and adaptive immune systems
Authors:	Waqas Ahmad (P73188)
Supervisor:	Ibrahim Jantan, Prof. Dr.
Keywords:	Tinospora crispa Immune system Dissertations, Academic -- Malaysia
Issue Date:	25-Jul-2017
Description:	Tinospora crispa (L) Hook. f. & Thomson has been numerously used as folkloric medicine for the treatment of various diseases due to its versatile pharmacological activities. However, the mechanistic effect of TC on the immune system was unexplored. The present study was aimed to investigate the immunomodulatory effects of 80% ethanol extract of T. crispa (TCE) and its chemical constituents using in vitro and in vivo models. TCE was qualitatively and quantitatively analyzed using a validated reversed-phased high performance liquid chromatography method while its chemical constituents were isolated by various chromatographic methods and identified by spectroscopic techniques. The in vitro assessment was performed by evaluating the immunomodulatory potential of TCE and its isolates using RAW 264.7 macrophages, by assessing their effects on chemotaxis, phagocytosis, production of inflammatory mediators such as reactive oxygen species (ROS), nitric oxide (NO), and pro inflammatory cytokines which include tumor necrosis factor-α (TNF-α), interleukin (IL)-1β, interleukin-6 (IL-6) and chemokine (MCP-1). In addition, the effects of TCE and its isolates on the mRNA expression of inducible nitric oxide synthase (iNOS), pro-inflammatory cytokines (IL-1β, TNF-α and IL6) and chemokine (MCP-1) were also evaluated using RT-PCR. The in vivo immunomodulatory prospects of TCE on cellular and humoral components of the immune response were investigated using Wistar Kyoto rats and Balb/c mice. In Wistar Kyoto rat model, the effect of TCE was examined by evaluating chemotactic and phagocytic activities of neutrophils isolated from the blood. To further elucidate the mechanistic approach, its effects on the proliferation of T- and B-lymphocytes, T lymphocytes subsets (CD4+ and CD8+) and the secretion of Th1 and Th2 cytokines were also monitored. Similarly, the immunomodulatory effects of TCE in Balb/c mice were investigated on cellular immune response by evaluating NO production, peritoneal macrophages phagocytosis and delayed type hypersensitivity (DTH), whereas humoral immune response was evaluated through measurement of serum immunoglobulin (IgG and IgM) and serum lysozyme levels. The chemical constituents isolated from TCE were N-formylannonain, N-formylnornuceferine, magnoflorine, lysicamine, syringin and 1-octacosanol. Results obtained demonstrated that TCE significantly enhanced the NO production and improved phagocytic activity of RAW 264.7 macrophages. Among the isolated compounds, magnoflorine showed remarkable inducing effects on the phagocytic activity, ROS and NO production as well as the secretion of IL-1β, TNF-α, IL6 and MCP-1 at the transcriptional and translational levels. In contrast, syringin potently reduced the phagocytic activity, ROS and NO production and secretion of IL-1β, TNF-α, IL6 and MCP-1 at the transcriptional and translational levels. The in vivo studies revealed that the chemotactic and phagocytic activities of the neutrophils extracted from TCE-treated rats were significantly increased as compared to the control group. The increasing trend was also observed in T and Blymphocytes proliferation. Besides cell mediated immune response, the levels of Th1 (TNF-α, IL-2 and IFN-γ) and Th2 (IL-4)-mediated cytokines were significantly increased in the rats exposed to TCE compared to the control group. TCE considerably improved the peritoneal macrophages' ability to engulf FITC-labeled E. coli and promoted NO production in a dose-dependent manner. TCE also markedly potentiated the SRBSinduced swelling rate of mice paw in DTH and significantly enhanced the level of serum immunoglobulins. As compared to the control group, serum lysozyme level and myeloperoxidase (MPO) activity were also significantly higher in TCE-treated groups. These findings suggested that T. crispa possessed promising immunostimulatory activities and stimulated the innate and adaptive arms of the immune response. Thus, T. crispa has potential to be developed and used as natural immunostimulant and may be useful for improvement of immune-related disorders.,Ijazah Doktor Falsafah
Pages:	202
Call Number:	QW504.W252i 2017 9
Publisher:	UKM, Kuala Lumpur
Appears in Collections:	Faculty of Pharmacy / Fakulti Farmasi

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