Archana Saxena

Diarylheptanoids Rich Fraction of Alnus nepalensis Attenuates Malaria Pathogenesis: In‐vitro and In‐vivo Study

Diarylheptanoids from Alnus nepalensis leaves have been reported for promising activity against filariasis, a mosquito‐borne disease, and this has prompted us to investigate its anti‐malarial and safety profile using in‐vitro and in‐vivo bioassays. A. nepalensis leaf extracts were tested in‐vitro against chloroquine‐sensitive Plasmodium falciparum NF54 by measuring the parasite specific lactate dehydrogenase activity. Among all, the chloroform extract (ANC) has shown promising anti‐plasmodial activity (IC50 8.06 ± 0.26 µg/mL). HPLC analysis of ANC showed the presence of diarylheptanoids. Efficacy and safety of ANC were further validated in in‐vivo system using Plasmodium berghei‐induced malaria model and acute oral toxicity in mice. Malaria was induced by intra‐peritoneal injection of P. berghei infected red blood cells to the female Balb/c mice. ANC was administered orally at doses of 100 and 300 mg/kg/day following Peters 4 day suppression test. Oral administration of ANC showed significant reduction of parasitaemia and increase in mean survival time. It also attributed to inhibition of the parasite induced pro‐inflammatory cytokines as well as afford to significant increase in the blood glucose and haemoglobin level when compared with vehicle‐treated infected mice. In‐vivo safety evaluation study revealed that ANC is non‐toxic at higher concentration. Copyright

Diarylheptanoids from Alnus nepalensis attenuates LPS-induced inflammation in macrophages and endotoxic shock in mice.

Diarylheptanoids, a group of plant secondary metabolites are increasingly recognized as potential therapeutic agents. The aim of study was to ascertain the anti-inflammatory profile of diarylheptanoids from Alnus nepalensis against lipopolysaccharide (LPS)-induced inflammation in macrophages and endotoxic shock in mice. Extracts prepared from dried leaves of A. nepalensis using standard solvents were tested against LPS-induced inflammation in macrophages. Among all, butanol extract (ANB) has shown most significant inhibition of pro-inflammatory cytokines without any cytotoxicity. HPLC analysis of ANB showed the presence of diarylheptanoids. The diarylheptanoids were further isolated and tested in-vitro for anti-inflammatory activity. Treatment of isolated diarylheptanoids (HOG, ORE and PLS) was able to reduce the production and mRNA level of pro-inflammatory cytokines (TNF-α and IL-6). Furthermore, we demonstrated that it inhibited the expression of NF-kB protein in LPS-induced inflammation in macrophages. In-vivo efficacy and safety profile of ANB revealed that oral treatment of ANB was able to improve the survival rate, and inhibited the production of pro-inflammatory cytokines in serum, attenuated vital organ injury in a dose dependent manner without any toxic effect at higher dose in mice. The results suggest that diarylheptanoids from A. nepalensis can be considered as potential therapeutic candidates for the management of inflammation related diseases.

Flavonoids Rich Fraction of Citrus limetta Fruit Peels Reduces Proinflammatory Cytokine Production and Attenuates Malaria Pathogenesis

Exploration of possible pharmacological effects along with characterisation of the bioactive compounds present in peels may have a key role in converting the fruit waste materials into therapeutic value added products. Extracts prepared from the Citrus limetta fruit peels were studied for antioxidant and anti- inflammatory activity using in-vitro bioassays. Among all, ClEt an ethanol extract of Citrus limetta fruit peels has shown promising anti-oxidant and anti-inflammatory activity. ClEt was further validated to ensure its safety evaluation at 2000mg/kg and anti-malarial efficacy at 100, 250, 500 mg/kg body weight with special reference to inflammatory mediators involved in malaria pathogenesis. In-vivo study revealed that ClEt was safe at higher dose and showed promising anti-malarial activity by inhibiting the parasitaemia and inflammatory mediators (IFN-γ, TNF-α, IL-6) involved in malaria pathogenesis, able to improve the haemoglobin and glucose level and increase the survival time. Chemical fingerprint of ClEt revealed the presence of flavonoids. Results suggested the suitability of ClEt, a flavonoid rich fraction of Citrus limetta fruit peels as a candidate for further investigation towards the management of malaria pathogenesis.

Beneficial health applications and medicinal values of Pedicularis plants: A review

Pedicularis plants (Orobanchaceae), popularly known as lousewort, are found in Asia, Europe, and North America, and have been used in Sowa-Rigpa, the Himalayan art of healing and a traditional system of medicine for treating various ailments in humans. A comprehensive compilation on this valuable medicinal plant is not available, however. The present extensive review provides insight into the salient medicinal properties of Pedicularis plants with respect to various health issues and diseases. Our previous studies on Pedicularis plants from the Changthang region of Ladakh (India) and research advances leading to new developments in this field have prompted this review. The information presented here has been compiled and analyzed from authenticated published resources available on Medline, Pubmed, Pubmed Central, Science Direct, and other scientific databases. The Pedicularis genus consists of approximately 600 species (83 of which are found in India), with commonly reported species being Pedicularis longiflora Rudolph, P. bicornuta Klotzsch, P. oederi Vahl, P. cheilanthifolia, and P. pectinata. The major phytoconstituents of the Pedicularis sp. are phenols, phenylethanoids, phenylpropanoids, flavonoids, iridoids, lignans, and alkaloids, among others. The existing literature highlights that these compounds possess antioxidant, immunomodulatory, anti-inflammatory, antidiabetic, antibacterial, antifungal, analgesic, antitumor, hepatoprotective, neuroprotective, muscle-relaxing, antifatigue, diuretic, antipyretic, antithrombus, antihemolysis, and DNA-repairing properties. This medicinal herb is used in the treatment of leucorrhoea, fevers, sterility, rheumatism, general debility, collapse, and urinary problems, and for revitalizing the blood circulation, improving digestion, and maintaining vitality. This review emphasizes the various medicinal aspects of Pedicularis sp. plants containing a variety of phytoconstituents. Besides phenols, terpenoids, flavonoids, lignans, tannins, iridoid, and phenylpropanoid glycosides are among the active constituents responsible for multiple health effects. However, further extensive research is required to characterize the various phytoconstituents of Pedicularis to explore their modes of action at a molecular level and identify other beneficial applications that can exploit the tremendous medicinal potential of this important herb.

Advances in Boerhaavia diffusa hairy root technology: a valuable pursuit for identifying strain sensitivity and up-scaling factors to refine metabolite yield and bioactivity potentials

The present study reports the Agrobacterium rhizogenes-mediated hairy root induction in of an ethno-medicinally significant herb–Boerhaavia diffusa L., for elucidating the underlying competence regarding its biosynthetic (i.e. boeravinone B and eupalitin) and bioactivity (antibacterial, antioxidant and anti-inflammatory) potentials. Host plant-specific receptiveness towards A. rhizogenes strains and disparity in compatibility threshold of leaf and nodal explants were evident. Only leaf explants responded, attaining hairy root induction with the ATCC 15834 followed by A4 and SA79 strains in reducing order of transformation efficiency. The growth behaviours differed amongst independent rhizoclones, and two clones of A4 (RBH) and ATCC 15834 (RBT8) origin demonstrated higher growth potentials. Polymerase chain reaction amplification of rol genes confirmed their transformed nature. Optimization of the appropriate solvent and reverse phase high-performance liquid chromatography parameters relating to the targeted metabolite production in the selected RBH and RBT8 clones revealed higher accumulation of eupalitin with the RBH clone having the best result of 1.44 times greater yield over the control root. Compared to the selected rhizoclones, the control roots however showed higher boeravinone B content. Devising a modified “stirred-tank” reactor through equipping with marine impellers and ring spargers facilitated high-density RBH root biomass yield with 6.1-fold and 1.15-fold yield increment of the boeravinone B and eupalitin respectively compared to shake-flask cultures. Considering the control roots, the RBH clone revealed analogous antioxidant/antibacterial activities with improved anti-inflammatory potential. The hairy root mediated higher production of boeravinone B and eupalitin could be achieved for the first time in bioreactor.

Antimalarial activity of phytol derivatives: in vitro and in vivo study

Severe malaria is one of the leading causes of mortality among children and pregnant woman globally. Resistance development against the frontline antimalarial drugs has created an alarming situation, which requires intensive drug discovery to develop effective, affordable, and accessible antimalarial agents possessing novel modes of action. As a part of our drug discovery program for antimalarial agents from plants, phytol a very commonly occurring diterpene alcohol in the plant was investigated for its antimalarial potential. In vitro antiplasmodial activity against the chloroquine-sensitive Plasmodium falciparum NF54 by measuring the parasite specific lactate dehydrogenase (pfLDH), showed moderate activity (IC50 211.5 ± 0.93 µM). Further, phytol was chemically converted into thirteen derivatives, which were evaluated for their antiplasmodial potential. All the derivatives were moderate active, but among all the derivatives palmitoyl (PhY-3; IC50 12.13 ± 0.31 µM) has found most active without any cytotoxic effect on macrophage cells. PhY-3 was further validated in an in vivo system for its efficacy and safety profile in mice. Oral administration of PhY-3 showed significant reduction of parasitemia and increased mean survival time. It also attributed significant increase in blood glucose and hemoglobin level, when compared with vehicle-treated P. berghei infected mice without any toxic effect on normal mice at a higher dose. These findings confirm suitability of the phytol derivatives as new chemical entities for further investigation towards the management of malaria.

Essential oil from waste leaves of Curcuma longa L. alleviates skin inflammation

BackgroundCurcuma longa L. is an important industrial crop used by medicinal and cosmetic industries in the world. Its leaves are a waste material after harvesting rhizomes. The aim of the study was to evaluate the chemical and pharmacological profile of essential oil from waste leaves of Curcuma longa (EOCl) against skin inflammation.MethodsEOCl was subjected to gas chromatography (GC) analysis for identification of essential oil constituents and its anti-inflammatory evaluation through in vitro and in vivo models.ResultsChemical fingerprinting using GC and GC–MS analysis of EOCl revealed the presence of 11 compounds, representing 90.29% of the oil, in which terpinolene (52.88%) and α-phellandrene (21.13%) are the major components. In the in vitro testing EOCl inhibited the production of pro-inflammatory cytokines (TNF-α, IL-6, IL-1β) in lipopolysaccharide (LPS) and 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced inflammation in the human keratinocyte cell line (HaCaT). Topical application of EOCl produced anti-inflammatory effects by reducing ear thickness, ear weight and ameliorating the level of pro-inflammatory cytokines (TNF-α, IL-6, IL-1β) at protein and mRNA levels as well as regulating the overproduction of oxidative markers and restoring the histopathological damage in a TPA-induced mouse model of inflammation.ConclusionThese findings of topical anti-inflammatory properties of EOCl provide a scientific basis for medicinal use of this plant material against inflammatory disorders.Graphical Abstract

In vitro and in vivo immunomodulatory potential of Pedicularis longiflora and Allium carolinianum in alloxan-induced diabetes in rats

Pedicularis longiflora Rudolph (Orobanchaceae) and Allium carolinianum Linn (Alliaceae) are two important medicinal plants found in trans-Himalayan Changthang. The immunomodulatory potential of these plants has not been explored. In the present study, we evaluated the in vitro and in vivo immunomodulatory potential of P. longiflora and A. carolinianum in alloxan-induced diabetes in Wistar rats. The ethanol extracts of the aerial parts of P. longiflora and whole plant parts of A. carolinianum were used for studying the in vitro immunomodulatory activity using lymphocyte stimulation and cytokine release assays. For the in vivo study, 5 groups of 6 rats per group, including alloxan-induced diabetic and plant extract-treated rats, were evaluated for cell-mediated immune (CMI) and humoral immune (HMI) responses in a 42-day experimental trial using doses of 500mg/kg b.wt. for P. longiflora and 250mg/kgbwt. for A. carolinianum. For P. longiflora, the median effective dose was found to be 500mg/kg. The in vitro lymphocyte stimulation index for P. longiflora was significantly higher (1.73±0.04, p<0.05) than that for A. carolinianum (1.27±0.06). However, the release of transforming growth factor-β1 (TGF-β1, 15.63±1.00, p<0.05) by P. longiflora was significantly lower than that by A. carolinianum (21.61±1.19), suggesting a better immune response by P. longiflora than by A. carolinianum. P. longiflora significantly increased the ear thickness (53.12%), inflammatory cellular infiltration (200.00±11.42), and total leukocyte count (7.44±0.02) compared to A. carolinianum (47.57%, 165.83±3.96, and 7.01±0.01, respectively). P. longiflora significantly reduced the percentage of leukocytes with depolarized mitochondria (3.24±0.16%) and apoptosis (1.81±0.07%), and induced a better CMI response than A. carolinianum. Significantly (p<0.05) higher hemagglutination titer (28.37±0.80) and IgG production (6.43±0.34mg/mL) were observed in the P. longiflora-treated group than in the A. carolinianum-treated group (23.93±0.58 and 6.23±0.37mg/mL). Plasma tumor necrosis factor-α (TNF-α) and TGF-β1 levels, and nuclear factor-κB (NF-κB) expression were significantly (p<0.05) lower in the P. longiflora-treated group than in the A. carolinianum-treated group. This may be due to better HMI responses produced by P. longiflora than by A. carolinianum. This is the first study to show that P. longiflora ethanol extract has more potent in vitro and in vivo immunomodulatory activities than A. carolinianum, especially in alloxan-induced diabetic rats. However, further research is needed to identify the different molecular mechanisms involved in mediating this immunomodulatory response.