Characterization and active component identification of Premna herbacea roxb. root extract reveals anti-inflammatory effect and amelioration of imiquimod induced psoriasis via modulation of macrophage inflammatory response.

BACKGROUND: 1-2.88% of human populations are affected by psoriasis, one type of chronic inflammatory skin disease. Skin thickenings, erythema, scaling in skin are the most important symptoms of psoriasis. There are renewed interests amongst scientists in studying anti-inflammatory property of the plant extracts due to lower side effects and cost effectiveness. There are few reports suggesting anti-inflammatory activity of Premna herbacea roxb. but lacks systematic evaluation of these properties. METHODS: We, initially tested the anti-inflammatory activity of crude root methanolic extract in vitro, where it significantly reduced LPS generated ROS in splenic macrophages. We further tested the TLC and HPLC fraction in order to find active ingredient in Premna herbacea roxb. root extract that ameliorated the chronic inflammation of skin and performed GC-MS and LC-MS studies to identify active component. Upon finding significant anti-inflammatory effect of the crude root extract in vitro, We studied the efficacy of the Premna herbacea roxb. root extract in Imiquimod induced psoriasis like skin inflammation in male BALB/C mice that closely resembles human psoriasis. Immunophenotyping, Cytokine productions were observed by flow cytometry, status of gene expression was done by Real time PCR and nuclear co-localization was studied by confocal microscopy. RESULTS: We observed progressive increase in signs and symptoms of the disease in imiquimod treated diseased animals but the Premna herbacea roxb. Root Methanolic Extract (PHRME) reduced the thickening of the skin, redness and scaling in these animals. In our study, along with progression of the disease, the production of macrophages increases and with the application of PHRME, the percentage of macrophages have reduced. CONCLUSION: As per the previous Indigenous traditional knowledge regarding use of Premna herbacea roxb. against inflammatory disorder and lack of detail mechanistic study of the crude root extract prompted us to elucidate the efficacy of the root extract in vitro and in vivo psoriatic mice model. For the first time we have identified three putative bioactive active components (5­hydroxy-7-methoxyflavanone, 3-Hydroxy-7,8,2',3'-tetramethoxyflavone, 2,4',6'-trimethoxy chalcone) from Premna herbacea root methanolic extract (PHRME) and we suggest PHRME and purified active fractions influence NFκB and COX2 signaling pathway to suppress inflammatory conditions. All of the purified components show strong binding efficiency in our molecular docking analysis. Our study also suggests that Premna herbacea roxb. root extract may be explored as cost effective alternative for established treatment regimen as our study also indicates low side effect of the extract against pre-clinical psoriatic model.

Nanoscale Diamond-Based Formulation as an Immunomodulator and Potential Therapeutic for Lymphoma.

Integrative medicine practices, such as Ayurveda, are popular in India and many South Asian countries, yet basic research to investigate the concepts, procedures, and medical benefits of ayurvedic products has received little attention and is not fully understood. Here, we report a functional nanodiamond-based traditional Ayurvedic herbomineral formulation, Heerak Bhasma (Ayu_ND), for the treatment of solid tumors called Dalton's lymphoma generated in CD1 mice. Ayu_ND-mediated immunostimulation significantly reduces tumor cell proliferation and induces apoptosis aided by the active participation of dendritic cells. Immunomodulatory Ayu_ND treatment is highly immunostimulatory and drives dendritic cells to produce TNF-α. Treatment with Ayu_ND significantly reduces the tumor volume, inhibits metastasis in distant vascularized organs, and increases the life span of tumor-bearing animals compared with untreated littermates. These events were associated with elevated serum levels of the protective cytokines IFN-γ and TNF-α and downregulated the disease, exacerbating TGF-ß. Ayu_ND-mediated therapeutic success was also accompanied by the depletion of regulatory T cells and enhanced vaccine-induced T-cell immunity, guided by the restoration of the memory CD8+ T-cell pool and prevention of PD-1-mediated T cell exhaustion. The results provide a basis for further evaluation of ayurvedic formulations and drug efficacy in treating cancers.

α-Ketoglutarate Inhibits Thrombosis and Inflammation by Prolyl Hydroxylase-2 Mediated Inactivation of Phospho-Akt.

BACKGROUND: Phospho-Akt1 (pAkt1) undergoes prolyl hydroxylation at Pro125 and Pro313 by the prolyl hydroxylase-2 (PHD2) in a reaction decarboxylating α-ketoglutarate (αKG). We investigated whether the αKG supplementation could inhibit Akt-mediated activation of platelets and monocytes, in vitro as well as in vivo, by augmenting PHD2 activity. METHODS: We treated platelets or monocytes isolated from healthy individuals with αKG in presence of agonists in vitro and assessed the signalling molecules including pAkt1. We supplemented mice with dietary αKG and estimated the functional responses of platelets and monocytes ex vivo. Further, we investigated the impact of dietary αKG on inflammation and thrombosis in lungs of mice either treated with thrombosis-inducing agent carrageenan or infected with SARS-CoV-2. FINDINGS: Octyl αKG supplementation to platelets promoted PHD2 activity through elevated intracellular αKG to succinate ratio, and reduced aggregation in vitro by suppressing pAkt1(Thr308). Augmented PHD2 activity was confirmed by increased hydroxylated-proline and enhanced binding of PHD2 to pAkt in αKG-treated platelets. Contrastingly, inhibitors of PHD2 significantly increased pAkt1 in platelets. Octyl-αKG followed similar mechanism in monocytes to inhibit cytokine secretion in vitro. Our data also describe a suppressed pAkt1 and reduced activation of platelets and leukocytes ex vivo from mice supplemented with dietary αKG, unaccompanied by alteration in their number. Dietary αKG significantly reduced clot formation and leukocyte accumulation in various organs including lungs of mice treated with thrombosis-inducing agent carrageenan. Importantly, in SARS-CoV-2 infected hamsters, we observed a significant rescue effect of dietary αKG on inflamed lungs with significantly reduced leukocyte accumulation, clot formation and viral load alongside down-modulation of pAkt in the lung of the infected animals. INTERPRETATION: Our study suggests that dietary αKG supplementation prevents Akt-driven maladies such as thrombosis and inflammation and rescues pathology of COVID19-infected lungs. FUNDING: Study was funded by the Department of Biotechnology (DBT), Govt. of India (grants: BT/PR22881 and BT/PR22985); and the Science and Engineering Research Board, Govt. of India (CRG/000092).

Curcumin: the multi-targeted therapy for cancer regression.

Tumors are multifaceted; in fact, numerous things happen in synchrony to enable tumor promotion and progression. Any type of cancer is associated with the modification of 300-500 normal genes and characterized by the deregulation of cell signaling pathways at multiple steps leading to cancer phenotype. Thus a proper management of tumorigenesis requires the development of multi-targeted therapies. Several adverse effects associated with present day cancer therapies and the thirsts for multi-targeted safe anticancer drug instigate the use of natural polyphenol, curcumin. It appears to involve a blend of anti-carcinogenic, pro-apoptotic, anti-angiogenic, anti-metastatic, immunomodulatory and antioxidant activities. Also the molecular mechanisms implicated for the pleotropic activities of curcumin are diverse and seem to involve a combination of cell signaling pathways at multiple levels of tumorigenesis. Being a potent scavenger of reactive oxygen species, curcumin also ameliorates systemic toxicity in tumor-bearer. Taken together, by placing particular emphasis on molecular basis of tumor promotion and progression this review summarizes the anti-cancer properties of curcumin that may be exploited for successful clinical cancer prevention.

Black tea-induced decrease in IL-10 and TGF-beta of tumor cells promotes Th1/Tc1 response in tumor bearer.

Several lines of evidence support that impairment of host immune function by tumor may be related to several strategies of tumor escape from immunosurveillance. We found that in Ehrlich's ascites carcinoma (EAC)-bearing mice, the tumor cells secrete immunosuppressive cytokines, transforming growth factor beta (TGF-beta) and interleukin-10 (IL-10) that induce a general T helper cells type 2 (Th2) dominance dampening the T cytotoxic cells type 1 (Tc1) population. Interestingly, black tea at the antitumor dose of 2.5% significantly reduced TGF-beta and IL-10 in tumor cells in vivo, thereby preventing Th2 dominance in the tumor bearers and initiating a Th1/Tc1 response. Thus, apart from its anticancer activity, this popular beverage also rejuvenates cancer immunosurveillance by modulating cytokine profiles and establishing Th1/Tc1 dominance in the tumor-bearing host.

Black tea-induced amelioration of hepatic oxidative stress through antioxidative activity in EAC-bearing mice.

It is recognized that during cancer, the disease itself as well as many of the anticancer drugs in use produce undesirable side effects such as hepatotoxicity. We have already demonstrated the antitumor and immunorestoring effects of black tea. Here we report Ehrlich's ascites carcinoma (EAC)induced hepatotoxicity and its protection by antitumor dose of black tea in mice. Hepatotoxicity was adjudged by liver histopathology and by measurement of plasma level of alkaline phosphatase (ALP). An attempt to delineate the underlying mechanisms revealed tumor-induced generation of reactive oxygen species (ROS) on one hand and depression in antioxidants that neutralize ROS, i.e., superoxide dismutase (SOD), catalase, reduced glutathione (GSH), and glutathione-S-transferase (GST), on the other. As a result, lipid peroxidation, which leads to damage of host cell components, was increased. Treatment with antitumor dose of black tea could replenish the host's antioxidant system and regress cancer-induced ROS significantly, thereby protecting the host's liver from lipid peroxidation and subsequent degeneration. Thus, unlike many other anticancer agents, black tea not only has antitumor and immunorestoring properties, but it also protects host liver from tumor-induced toxicity. These results thus raise the possibility of inclusion of black tea in a successful therapeutic regimen against cancer.

Tumor-induced thymic involution via inhibition of IL-7R alpha and its JAK-STAT signaling pathway: protection by black tea.

Down-regulation of cell-mediated immune functions occurring at late stages of cancer may be related to the thymic involution since thymus is the major site of T cell maturation, proliferation, and differentiation. We found that in Ehrlich's ascites carcinoma (EAC)-bearing mice there was profound depletion of CD4+ and CD8+ cells in peripheral blood with severely damaged thymus on 21st day of tumor inoculation. However, treatment with black tea at an antitumor dose of 2.5% significantly reduced such depletion and protected the thymus considerably from tumor onslaught. A search for the underlying mechanism revealed EAC-induced IL-7Ralpha down-regulation, inhibition of JAK3 and STAT5 phosphorylation, and decrease in Bcl-2/Bax ratio in thymocytes that finally led to thymocyte apoptosis in one hand and T cell maturation block on the other. Interestingly, black tea treatment prevented IL-7Ralpha down-regulation and protected the signaling cascade through JAK-STAT thereby inhibiting tumor-induced thymic apoptosis and ensuring proper functioning of this organ in tumor-bearing host.