Andrographolide attenuates complete freund's adjuvant induced arthritis via suppression of inflammatory mediators and pro-inflammatory cytokines.

ETHNOPHARMACOLOGICAL RELEVANCE: Traditional plant-derived medicines have enabled the mankind in curing the wide spectrum of diseases throughout the ages. Andrographis paniculata (Burm.f.) Nees, is one of the traditional plant used as a folk medicine for the management of inflammation, arthritis, viral-bacterial infections and other ailments in India, China, Malaysia and other South-East Asian countries. Its major bioactive compound; andrographolide, a diterpenoid, also exerts cytoprotective properties and is reported to be effective in neuroprotection, hepatoprotection, etc. AIM: The study is aimed to explore the role of andrographolide in treatment of complete freund's adjuvant (CFA) induced arthritis. MATERIALS AND METHODS: The influx of immune cells, release of pro-inflammatory cytokines and subsequent accumulation of synovial fluid (swelling) and pain manifest into the disease. The present study used CFA induced Balb/c mice model and treated them intraperitoneally with andrographolide and dexamethasone (used as a positive control) on alternate days for six days. After 6 days, blood and peritoneal macrophages were collected to evaluate the expression of various arthritic markers and paw edema was measured on all days. RESULTS: The in vitro and ex vivo experiments showed that andrographolide treated animal group had reduced paw edema, cell cytotoxicity and nitric oxide production than dexamethasone treated animal group. Further, the study revealed the mechanistic role of andrographolide in treatment of arthritis by suppressing battery of molecules like COX-2, NF-κB, p-p38, CD40, TNF-α, IL-1ß and IL-6 involved in arthritis. CONCLUSION: The study showed the potent anti-arthritic effects of andrographolide and warrants further investigations on andrographolide for the development of safe and effective anti-arthritic drug.

Improvement in resilience and stress-related blood markers following ten months yoga practice in Antarctica.

BACKGROUND: Wintering is associated with distress to humans who work in the isolated and confined environment of Antarctica and yoga has been proved helpful for coping with stress. Therefore, a study was conducted on 14 winter expedition members of Indian Scientific Antarctic Expedition (2016) to find out the effects of yoga on stress-related markers. METHODS: Participants were divided into yoga, and control (non-yoga) groups. The yoga group practiced yoga for 10 months (from January to October 2016) daily in the morning for an hour. The Resilience test questionnaire was administrated at baseline and endpoint of the study. Blood samples were collected during the study at different intervals for the estimation of 8-hydroxydeoxyguanosine (8-OHdG), brain-derived neurotrophic factor (BDNF), serotonin and cortisol using ELISA. RESULTS: A trend of improvement was observed in the resilience test score in the yoga group. From January to October, 8-OHdG serum values in the yoga group declined by 55.9% from 1010.0 ± 67.8 pg/mL to 445.6 ± 60.5 pg/mL (Mean ± SD); in the control group, the decline was 49.9% from 1060.4 ± 54.6 pg/mL to 531.1 ± 81.8 pg/mL. In serotonin serum levels in the yoga group, there was a 3.1% increase from 6.4 ± 1.6 ng/mL to 6.6 ± 0.4 ng/mL while no increase was noticed in the control group. Cortisol values in the yoga group decreased by 19.9% from 321.0 ± 189.6 ng/mL to 257.1 ± 133.8 ng/mL; in the control group it increased by 2.8% from 241.2 ± 51.8 ng/mL to 247.8 ± 90.9 ng/mL. CONCLUSIONS: It could be concluded from the present study that following 10 months yoga practice may be useful for better resilience and management of stress-related blood markers for the polar sojourners.

Design and validation of Integrated Yoga Therapy module for Antarctic expeditioners.

BACKGROUND: Extreme environments are inherently stressful and are characterized by a variety of physical and psychosocial stressors, including, but not limited to, isolation, confinement, social tensions, minimal possibility of medical evacuation, boredom, monotony, and danger. Previous research studies recommend adaptation to the environment to maintain optimal function and remain healthy. Different interventions have been tried in the past for effective management of stress. Yoga practices have been shown to be beneficial for coping with stress and enhance quality of life, sleep and immune status. OBJECTIVE: The current article describes preparation of a Yoga module for better management of stressors in extreme environmental condition of Antarctica. MATERIALS AND METHODS: A Yoga module was designed based on the traditional and contemporary yoga literature as well as published studies. The Yoga module was sent for validation to forty experts of which thirty responded. RESULTS: Experts (n = 30) gave their opinion on the usefulness of the yoga module. In total 29 out of 30 practices were retained. The average content validity ratio and intra class correlation of the entire module was 0.89 & 0.78 respectively. CONCLUSION: A specific yoga module for coping and facilitating adaptation in Antarctica was designed and validated. This module was used in the 35th Indian Scientific expedition to Antarctica, and experiments are underway to understand the efficacy and utility of Yoga on psychological stress, sleep, serum biomarkers and gene expression. Further outcomes shall provide the efficacy and utility of this module in Antarctic environments.

Evaluation of anti-dengue activity of Carica papaya aqueous leaf extract and its role in platelet augmentation.

Dengue disease is characterized by a marked decrease in platelet count, which is life threatening. In the present study, we investigated the antiviral activity of an aqueous extract of Carica papaya leaves (PLE) against dengue virus (DENV) and its effect on platelet augmentation. The anti-dengue activity of PLE in DENV-infected THP-1 cells was examined by immunoblotting and flow cytometry. The effect of PLE on erythrocyte damage was investigated using hemolytic and anti-hemolytic assays. Virus-infected THP-1 cells were assayed for IFN-α secretion. The effect of PLE on platelet augmentation in rats with cyclophosphamide-induced thrombocytopenia was also investigated. The platelet count of blood from the retro-orbital plexus of rats was determined on the 1st, 4th, 7th, 11th and 14th day of study. On the 14th day, the rats were sacrificed for histopathological examination of the liver, kidney and spleen. Plasma of thrombocytopenic rats was tested for thrombopoietin (TPO) and IL-6 secretion. The data suggest that PLE significantly decreases the expression of the envelope and NS1 proteins in DENV-infected THP-1 cells. A marked decrease in intracellular viral load upon PLE treatment confirmed its antiviral activity. This also resulted in a significant decrease in erythrocyte damage and hydrogen-peroxide-induced lipid peroxidation. A significant increase in the number of platelets was found in thrombocytopenic rats treated with PLE, along with an increase in IL-6 and TPO levels. These findings suggest that PLE can potentially be used as an antiviral agent, as it helps in platelet augmentation and exhibits antiviral activity against DENV.

A novel in vivo adjuvant activity of kaempferol: enhanced Tbx-21, GATA-3 expression and peritoneal CD11c+MHCII+ dendritic cell infiltration.

OBJECTIVE: Kaempferol, a natural flavonol present in various traditional medicinal plants, is known to possess potent anti-inflammatory properties. This study was designed to study the adjuvant effect of kaempferol administration along with ovalbumin antigen (K + O) in balb/c mice. METHODS: Mice were immunized with kaempferol (100 and 50 mg/kg body weight) without or with ovalbumin (20 µg/mouse). After priming, booster was administered on day 21. Antigen specific IgG titers and its subtypes, on day 28, were estimated by indirect ELISA. Effect of kaempferol administration on CD11c+MHCII+ peritoneal dendritic cells was studied by flow cytometry. Expression levels of proteins Tbx21, GATA-3, BLIMP-1, Caspase-1 and Oct-2 were studied by western blotting. LPS activated IL-1ß production by peritoneal cells of immunized mice was estimated by sandwich ELISA. RESULTS: Ovalbumin specific IgG, IgG1 and IgG2a antibody titers in sera samples of K + O immunized mice increased significantly (p < .01) as compared to controls. The enhanced Th1 and Th2 immune response in K + O immunized mice was also supported by the increased expression of Tbx21 and GATA-3 transcription factors in splenocytes. This corroborated with increased BLIMP-1 and Oct-2 protein expression. Kaempferol increased the infiltration of peritoneal CD11c+MHCII+ dendritic cells but failed to enhance LPS activated IL-1ß by peritoneal macrophages and suppressed caspase-1 protein expression as compared to that in ovalbumin immunized mice. CONCLUSION: Present study strongly demonstrates the novel adjuvant activity of kaempferol in vivo and its potential as an immunostimulatory agent.

Anti-inflammatory activity of the functional groups present in Hippophae rhamnoides (Seabuckthorn) leaf extract.

ETHNO PHARMACOLOGICAL RELEVANCE: The study explores the anti-inflammatory activity of components present in fractions obtained from leaves of Hippophae rhamnoides in mouse peritoneal macrophages. AIM OF THE STUDY: Immunomodulators salvage the immune response by enhancing or reducing its capacity to the required level. Plant extracts are extensively used as immunomodulators because of their easy availability, simple methods of preparation and minimum side effects with maximum efficacy. MATERIALS AND METHODS: The present study was conducted to assess the immunomodulatory activities of phyto constituents present in Seabuckthorn leaves. The aqueous-alcoholic leaf extract was subjected to successive and parallel extraction in the presence of polar and non-polar solvents for fractionation of compounds. Based on the yield, three fractions were selected viz. parallel methanol (PM), successive chloroform (SC) and successive methanol (SM) and screened for in vitro immunomodulatory activities. Peritoneal macrophages were isolated from Balb/c mice and cultured with or without LPS to evaluate the immunomodulatory effect of the three fractions on cell viability, hemolytic activity, nitric oxide (NO) production, cytokine levels, iNOS and COX-2 expressions. RESULTS: The results revealed that none of the three fractions induced hemolysis. Cells treated with PM fraction significantly suppressed LPS-induced NO production and pro-inflammatory cytokines such as TNF-α, IL-6 and IFN-γ as compared to SC and SM treatment. The iNOS and COX-2 expressions were also significantly reduced after treatment with PM fraction. CONCLUSIONS: The decrease in LPS-induced NO production, pro-inflammatory cytokine secretion, iNOS and COX-2 expression signifies anti-inflammatory properties of PM fraction containing tannins, proteins and carbohydrate groups. Hence, this plant-derived immunomodulator can be used as a therapeutic agent in inflammatory diseases.

Andrographolide - A promising therapeutic agent, negatively regulates glial cell derived neurodegeneration of prefrontal cortex, hippocampus and working memory impairment.

Over activation of glial cell derived innate immune factors induces neuro-inflammation that results in neurodegenerative disease, like working memory impairment. In this study, we have investigated the role of andrographolide, a major constituent of Andrographis paniculata plant, in reduction of reactive glial cell derived working memory impairment. Real time PCR, Western bloting, flow cytometric and immunofluorescence studies demonstrated that andrographolide inhibited lipopolysaccharide (LPS)-induced overexpression of HMGB1, TLR4, NFκB, COX-2, iNOS, and release of inflammatory mediators in primary mix glial culture, adult mice prefrontal cortex and hippocampus region. Active microglial and reactive astrocytic makers were also downregulated after andrographolide treatment. Andrographolide suppressed overexpression of microglial MIP-1α, P2X7 receptor and its downstream signaling mediators including-inflammasome NLRP3, caspase1 and mature IL-1ß. Furthermore, in vivo maze studies suggested that andrographolide treatment reversed LPS-induced behavioural and working memory disturbances including regulation of expression of protein markers like PKC, p-CREB, amyloid beta, APP, p-tau, synapsin and PSD-95. Andrographolide, by lowering expression of pro apoptotic genes and enhancing the expression of anti-apoptotic gene showed its anti-apoptotic nature that in turn reduces neurodegeneration. Morphology studies using Nissl and FJB staining also showed the neuroprotective effect of andrographolide in the prefrontal cortex region. The above studies indicated that andrographolide prevented neuroinflammation-associated neurodegeneration and improved synaptic plasticity markers in cortical as well as hippocampal region which suggests that andrographolide could be a novel pharmacological countermeasure for the treatment of neuroinflammation and neurological disorders related to memory impairment.

Augmentation of humoral and cellular immunity in response to Tetanus and Diphtheria toxoids by supercritical carbon dioxide extracts of Hippophae rhamnoides L. leaves.

Hippophae rhamnoides L. commonly known as Seabuckthorn (SBT), a wild shrub of family Elaegnacea, has extensively used for treating various ailments like skin diseases, jaundice, asthma, lung troubles. SBT leaves have been reported to possess several pharmacological properties including immunomodulatory, antioxidant, anti-inflammatory, antimicrobial and tissue regeneration etc. The present study was undertaken to evaluate the adjuvant property of supercritical carbon dioxide extracts (SCEs 300ET and 350ET) of SBT leaves in balb/c mice immunized with Tetanus and Diphtheria toxoids. The dynamic changes in the immune response were measured in terms of humoral and cell-mediated immune responses. We have seen the effect of SCEs on immunoglobulin subtypes and secondary immune response generation. In addition, the effect of SCEs on antigen specific cellular immunity was evaluated. Our results show that SCEs 300ET and 350ET significantly enhanced antibody titers in response to both TT and DT antigens. The secondary immune response generated was significantly increased in case of TT immunized animals. SCEs also enhanced cytokine levels (IFN-γ, IL-4, TNF-α and IL-1ß) and increased lymphoproliferation. Besides, both SCEs did not show any toxic effects. Therefore, the study suggests that SCEs are safe and have potent immunostimulatory activity and hence, seems to be a promising balanced Th1 and Th2 directing immunological adjuvant for various veterinary as well as human vaccines.

Golden root: A wholesome treat of immunity.

Rhodiola is native to the high altitude regions of Asia, Europe and Northern Hemisphere. It has a long history of use as a medicinal plant in various ailments, boosting immunity, increasing energy and mental capacity. It is also known as "Adaptogen" to help the body to adapt and resist stress. The part of the plant, which is used for medicinal values, is rhizome, which is an underground stem. The rhizome contains mainly salidroside, rosin, rosavin and p-tyrosol. There are many studies, which have reported the effects of Rhodiola spp. on different organs and health conditions. In this review, we have selected the articles from Pubmed and Google Scholar from year 1992-2016 to report the effects of Rhodiola spp. and their role in curtailing various diseases and stress. The present review emphasizes the medicinal and therapeutic applications of Rhodiola spp. on different experimental models. Overall conclusion is that Rhodiola spp. has immense therapeutic potential and hence, this review would give impetus to new research for the development of Rhodiola based herbal nutraceuticals as well as pharmaceuticals.

Broad-spectrum antiviral properties of andrographolide.

Andrographolide, a diterpenoid, is known for its anti-inflammatory effects. It can be isolated from various plants of the genus Andrographis, commonly known as 'creat'. This purified compound has been tested for its anti-inflammatory effects in various stressful conditions, such as ischemia, pyrogenesis, arthritis, hepatic or neural toxicity, carcinoma, and oxidative stress, Apart from its anti-inflammatory effects, andrographolide also exhibits immunomodulatory effects by effectively enhancing cytotoxic T cells, natural killer (NK) cells, phagocytosis, and antibody-dependent cell-mediated cytotoxicity (ADCC). All these properties of andrographolide form the foundation for the use of this miraculous compound to restrain virus replication and virus-induced pathogenesis. The present article covers antiviral properties of andrographolide in variety of viral infections, with the hope of developing of a new highly potent antiviral drug with multiple effects.

A pharmacological investigation of Hippophae salicifolia (HS) and Hippophae rhamnoides turkestanica (HRT) against multiple stress (C-H-R): an experimental study using rat model.

Hippophae salicifolia (HS) and Hippophae rhamnoides turkestanica (HRT) are abundantly found species of Hippophae in Himalayan region of India. As these plants thrive under extreme climatic conditions, it is suspected that these plants must have a unique adaptogenic property against high-altitude stress. To keeping these views in our mind, the present study was planned to evaluate the mechanism of action of aqueous extract of HS and aqueous extract of HRT against multiple stress [cold-hypoxia-restraint (C-H-R)] for their adaptogenic activity. The present study reported the adaptogenic activity of HS in facilitating tolerance to multiple stress, CHR in rats. Pre-treatment with aqueous extract of HS significantly attenuated reactive oxygen species (ROS) production, protein oxidation, and lipid peroxidation and also showed role in maintaining antioxidant status as similar to control rats. Since protein oxidation was decreased by pre-treatment of HS, protein homeostasis was also sustained by regulation of heat shock proteins (HSP70 and HSP60). Interestingly, heme oxygenase-1 (HO-1), Vascular Endothelial Growth Factor (VEGF), and nitric oxide (NO) level was also increased in HS pre-treated rats depicted its adaptogenic activity against multiple stress, CHR. Conclusively, aqueous extract of HS could use an adaptogen for high altitude-associated multiple stress (CHR).

Rhodiola inhibits dengue virus multiplication by inducing innate immune response genes RIG-I, MDA5 and ISG in human monocytes.

Recognition of virus infection by retinoic acid-inducible gene (RIG) I and melanoma differentiation-associated protein (MDA) 5, which are RNA helicases, and interferon-stimulated gene (ISG) 15 activates cascades of signal transduction pathways leading to production of type I interferons and proinflammatory cytokines that orchestrate the elimination of the viruses. However, it has been demonstrated that RNA-helicase-mediated innate immunity plays an essential role in defending the host from infection. In our efforts to identify plant-derived antivirals that selectively enhance ISG- and RNA-helicase-mediated antiviral immune responses, we identified a plant, rhodiola, that significantly promoted ISG, RIG-I and MDA 5 gene expression and an antiviral immune response against dengue virus (DENV) infection. Rhodiola induced interferon (IFN) ß and other cytokines, including IL-1ß, TNF-α, IL-6 and IL-8, in infected cells. It was also found that rhodiola upregulated phosphorylated eIF-2α, PKR and NF-kB in infected cells. In addition, the number of NK cells was also increased by rhodiola treatment in dengue-virus-infected human PBMCs. Treatment with a crude extract of rhodiola (RAE) resulted in effects in the 20 % range, which is similar to the magnitude of the same effects observed in DENV infections. Taken together, our results imply that rhodiola induces pharmacological modulation of RIG-I, MDA 5 and ISG signal transduction pathways in favor of the induction of a beneficial antiviral immune response against dengue virus, which can be a novel therapeutic strategy for management of infection.

Anti-cellular and immunomodulatory potential of aqueous extract of Rhodiola imbricata rhizome.

In the present study, we have evaluated the anti-cellular and immunomodulatory potential of aqueous extract of Rhodiola imbricata rhizome (RAE). Rhodiola extract inhibited the proliferation of human T cell lymphoma cell line EL-4 and erythroleukemic cell line HL-60. Furthermore, treatment of human peripheral blood mononuclear cells (hPBMCs) with lipopolysaccharide (LPS) and RAE suppressed regulated upon activation, normal T cell expressed and secreted (RANTES) production. However, number of TNF-α spots was increased in RAE treated hPBMCs. The reverse transcriptase polymerase chain reaction (RT-PCR) analysis of RAE treated rat splenocytes confirmed the up regulation of TLR-4 mRNA expression. Therefore, the present study concludes that RAE has potent immune boosting activity which might be useful in immunocompromised individuals.

Adjuvant effect of aqueous extract of Rhodiola imbricata rhizome on the immune responses to tetanus toxoid and ovalbumin in rats.

In the present study we have evaluated the immunopotentiating activity of Rhodiola aqueous extract (RAE) in rats. The efficacy of RAE was determined by using strong antigen tetanus toxoid (TT) and weak antigen Ovalbumin (OVA). The dynamic changes in humoral and cell-mediated immune response were measured. The results indicated that the TT specific immunoglobulin levels were significantly enhanced by RAE and were at par with complete Freund's adjuvant (CFA). The level of OVA induced antibody response was also enhanced by RAE. It was observed that TT and OVA in combination with CFA or RAE could evoke a significant delayed type hypersensitivity (DTH) response, confirming its potential to generate strong cell-mediated immunity (CMI). The anti-inflammatory or immunosuppressive effect of RAE was evaluated in adjuvant-induced arthritis model (AIA). RAE could not suppress the swelling response. Therefore, this study suggests that RAE has adjuvant/immunopotentiating activity in terms of humoral as well as cell-mediated immune response against strong antigen like TT and weak antigen like OVA.

Aqueous extract of Rhodiola imbricata rhizome stimulates Toll-like receptor 4, granzyme-B and Th1 cytokines in vitro.

Rhodiola imbricata is a medicinal plant, native to mountainous regions of Asia, parts of Europe, and the Arctic. Traditionally it is recommended to help combat fatigue and restore energy. It exhibits anti-stress, anti-cancer, and immunostimulatory activities. However, the effect of Rhodiola on immunological responses largely remains unknown. In this study, we have investigated the effect of aqueous extract of R. imbricata rhizome (RAE), on Toll-like receptor-4 (TLR-4) and intracellular granzyme-B expression in mouse splenocytes. Furthermore, TH1/TH2 cytokine profile was analyzed in RAE-treated human peripheral blood mononuclear cells (PBMCs) using multiplex flowcytomix kit. Our findings suggest that RAE induces TLR-4 expression and intracellular granzyme-B in treated splenocytes while RAE stimulated IL-1beta, IL-6, and TNF-alpha in human PBMCs. The present study suggests that RAE stimulates the innate immune pathway and has potent immunostimulatory activity, which can be used in modulating the immune system of immunocompromised individuals.

Effect of leaf extract of Seabuckthorn on lipopolysaccharide induced inflammatory response in murine macrophages.

Nitric oxide (NO) is synthesized in large quantities by activated inflammatory cells and has been demonstrated to be involved in the pathogenesis of acute and chronic inflammatory conditions. Seabuckthorn (SBT) has been used in traditional medicine systems for the treatment of various diseases like cardiovascular, pain relief, oral inflammation and promotion of tissue regeneration. The present study focuses on the effects of SBT leaf extract on NO production induced by lipopolysaccharide (LPS) in the murine macrophage cell line RAW 264.7. In addition, cell viability, free radical-scavenging activity and inducible nitric oxide synthase (iNOS) expression were also evaluated. Seabuckthorn leaf extract significantly inhibited the enhanced production of NO induced by LPS in a dose dependent manner. Treatment with SBT did not reduce cell viability at any dose used. The extract showed significant scavenging of NO radicals released by the NO donor. Treatment of macrophages with SBT leaf extract also caused a significant inhibition of iNOS activation. These observations suggest that the inhibition of net NO production by SBT leaf extract may be due to its scavenging activity and/or its inhibitory effects on iNOS activation. The study suggests that SBT leaf extract has significant anti-inflammatory activity and has potential for the treatment of inflammatory diseases.

Anti-inflammatory activity of Seabuckthorn (Hippophae rhamnoides) leaves.

Immunomodulatory activity of Seabuckthorn (SBT) leaf extract was evaluated in adjuvant induced arthritis (AIA) rat model. Inflammation was induced by injecting Complete Freund's Adjuvant (CFA) in the right hind paw of rats. SBT extract was administered intraperitoneally to treat the inflammation. The extent of inflammation and treatment response was evaluated by clinical analysis, scintigraphic visualization using technitium-99m-glutathione (Tc99m-GSH) and lymphocyte proliferation. Serial evaluation was carried out on days 1, 7, 14, 21 and 28 after creation of inflammation. The Tc99m-GSH uptake in the inflamed leg was compared with the normal contralateral leg of the same animal. The measurements were done by obtaining scintigraphic images using gamma camera and an online computer. Both qualitative and quantitative evaluation of radiotracer accumulation was considered to evaluate the anti-inflammatory response. The lymphocyte proliferation study revealed cellular immunosuppression during the early phase of the disease. Administration of SBT extract on the same day or 5 days prior to inflammatory insult into the joint, significantly reduced the inflammation as compared to the untreated animals in a dose dependent manner. These observations suggest that the SBT leaf extract has a significant anti-inflammatory activity and has the potential for the treatment of arthritis.

Immunomodulatory effects of agents of plant origin.

The immunomodulatory properties of amla (Emblica officinalis) and shankhpushpi (Evolvulus alsinoides) were evaluated in adjuvant induced arthritic (AIA) rat model. Injecting Complete Freund's Adjuvant (CFA) in right hind paw of the animals induced inflammation. The crude extracts of both the herbs were administered intraperitonially following a repeated treatment profile. The anti-inflammatory response of both the extracts was determined by lymphocyte proliferation activity and histopathological severity of synovial hyperplasia. Both the extracts showed a marked reduction in inflammation and edema. At cellular level immunosuppression occurred during the early phase of the disease. There was mild synovial hyperplasia and infiltration of few mononuclear cells in amla or shankhpushpi treated animals. The induction of nitric oxide synthase (NOS) was significantly decreased in treated animals as compared to controls. These observations suggest that both the herbal extracts caused immunosuppression in AIA rats, indicating that they may provide an alternative approach to the treatment of arthritis.