Stabilizing histamine release in gut mast cells mitigates peripheral and central inflammation after stroke.

Stroke is the most common cause of long-term disability and places a high economic burden on the global healthcare system. Functional outcomes from stroke are largely determined by the extent of ischemic injury, however, there is growing recognition that systemic inflammatory responses also contribute to outcomes. Mast cells (MCs) rapidly respond to injury and release histamine (HA), a pro-inflammatory neurotransmitter that enhances inflammation. The gut serves as a major reservoir of HA. We hypothesized that cromolyn, a mast cell stabilizer that prevents the release of inflammatory mediators, would decrease peripheral and central inflammation, reduce MC trafficking to the brain, and improve stroke outcomes. We used the transient middle cerebral artery occlusion (MCAO) model of ischemic stroke in aged (18 mo) male mice to investigate the role of MC in neuroinflammation post-stroke. After MCAO we treated mice with 25 mg/kg body weight of cromolyn (MC stabilizer) by oral gavage. Cromolyn was administered at 3 h, 10 h, 24 h and every 24 h for 3 days post-stroke. Three control groups were used. One group underwent a sham surgery and was treated with cromolyn, one received sham surgery with PBS vehicle and the third underwent MCAO with PBS vehicle. Mice were euthanized at 24 h and 3 days post-stroke. Cromolyn administration significantly reduced MC numbers in the brain at both 24 h and 3 days post-stroke. Infarct volume was not significantly different between groups, however improved functional outcomes were seen at 3 days post-stroke in mice that received cromolyn. Treatment with cromolyn reduced plasma histamine and IL-6 levels in both the 24-h and 3-day cohorts. Gut MCs numbers were significantly reduced after cromolyn treatment at 24 h and 3 days after stroke. To determine if MC trafficking from the gut to the brain occurred after injury, GFP+MCs were adoptively transferred to c-kit-/- MC knock-out animals prior to MCAO. 24 h after stroke, elevated MC recruitment was seen in the ischemic brain. Preventing MC histamine release by cromolyn improved gut barrier integrity and an improvement in stroke-induced dysbiosis was seen with treatment. Our results show that preventing MC histamine release possesses prevents post-stroke neuroinflammation and improves neurological and functional outcomes.

Evaluation of Pharmacological Potential of Miliusa nilagirica Bedd. Leaves Using In Vitro Antidiabetic and Antioxidant Assays.

Miliusa nilagirica, a rare tree species of Western Ghats, belongs to the Annonaceae family, a family with potential antioxidant and antidiabetic properties. This study is designed vividly to establish the relationship between the constituent phytochemicals and their hyperglycemic effects through the antioxidant traits of M. nilagirica in vitro. Phytochemical tests were conducted on dry powdered leaves and extracts of various methods to determine the existence of various constituents. The antidiabetic potential of leaf extracts was estimated by using the α-amylase inhibitory model and the antioxidant potential was estimated with various assays. The quantitative phytochemical screening of leaf parts shows the presence of carbohydrates (88.74 ± 0.65 mg GE/g sample), proteins (82.17 ± 2.52 mg BSAE/g sample), phenolics (40.44 ± 0.43 GAE/100 g), and flavonoids (66.05 ± 0.48 mg RE/g extract). Methanol extract of Soxhlet of M. nilagirica registered the strongest antioxidant activity in all assays, 75.66% inhibition (DPPH assay), 795.01 µmol/g (ABTS˙+ radical scavenging), 994.33 µmol/g (FRAP assay), 362.02 mg AAE/g extract (TAC assay), 47% inhibition (NO scavenging assay). In vitro α-amylase inhibition showed a highly noticeable reduction in ethyl acetate extract from Soxhlet (75.19%). HPLC and FTIR analyses on the extracts added strengths to the obtained results on the potentiality of M. nilagirica. From the results, it is evident that phytochemicals from M. nilagirica can be studied further, isolated, and incorporated as an alternative to synthetic supplements for hyperglycemia.

Efficacy of Trevesia palmata (Roxb. ex Lindl.) Vis. Extract on MG 63 cell lines and arthritis-induced animal models.

ETHNOPHARMACOLOGICAL RELEVANCE: Despite widespread use of herbal remedies for treating arthritis and osteosarcoma, many plants are still not pharmacologically evaluated for their efficacy. Contrary to many non-steroidal, immunosuppressants, antibiotics, and antineoplastic drugs that have adverse effects, phytotherapeutic compounds have promising benefits with fewer complications. In this study the unexplored Northeastern India indigenous plant Trevesia palmata (Roxb. ex Lindl.) Vis. used in traditional medicine to cure bone fractures is chosen for studying anti-proliferative and anti-rheumatic properties. AIM OF THE STUDY: This study designed to explore the polyphenolic composition, antioxidant, anti-inflammatory and anti-arthritic potential of T. palmata leaf extracts. Further, the cellular activity was studied using MG 63 osteoblast cell lines and pharmacologically evaluated using Complete Freund's Adjuvant (CFA) induced arthritic rat model. MATERIALS AND METHODS: In vitro free radical scavenging activity, anti-inflammatory and anti-arthritic activities of extracts were analyzed using standardized methods. The polyphenolic profiling and apoptosis inducing ability of T. palmata ethyl acetate (TPEA) extract on MG 63 osteoblast cell lines were analyzed. The in vivo pharmacological studies were carried out with low dose 250 mg/kg and high dose of 500 mg/kg of T. palmata. The biochemical and haematological parameters and in vivo antioxidant activity were evaluated for the control and treated groups. Radiological and histological study were done to understand the impact and penetration of inflammatory arthritis from tissues to joint bones. RESULTS: TPEA showed highest free radical scavenging activity (DPPH - 4.72 IC50, ABTS - 242.33 ± 6.81 mM TE/g extract), anti-inflammatory (40.04% inhibition of RBC lysis) and anti-arthritic activity (32.4% inhibition of protein denaturation) with the presence of gallic acid, catechin, caffeic acid, rutin, quercetin and naringenin. The TPEA extract inhibited cell proliferation of MG 63 osteoblast cells and induced apoptosis by arresting cell cycle at different phases. After acute toxicity studies the doses 250 mg/kg and 500 mg/kg were fixed and showed better results in CFA-induced arthritic animals. Thus, the extract phytoconstituents may have immense potential against chronic inflammation, joint ailments, bone cancer and arthritis which serves as a phytomedicine contrary to synthetic medications. CONCLUSIONS: The potential treatment of polyphenolic compounds in the T. palmata extract on osteosarcoma and arthritis was demonstrated from this study. Thus, cellular inflammatory infiltrates are significantly reduced in bone and joint tissues as well.

Ipomoea pes-tigridis L. extract accelerates wound healing in Wistar albino rats in excision and incision models.

ETHNOPHARMACOLOGICAL RELEVANCE: An annual herb, Ipomoea pes-tigridis L. (Convolvulaceae) is widely used for its anti-inflammatory and anti-spasmodic properties in traditional medicine. As well as treating wounds, fever, skin disorders, and other ailments, it is also used for other purposes. AIM OF THE STUDY: This study investigated polyphenolic content, antioxidant activity, RP-HPLC, wound healing, and antioxidant enzyme activity. In terms of I. pes-tigridis potential for healing wounds, there is no scientific data available. Hence this study is designed to use animal models to investigate the ethnopharmacological report. MATERIALS METHODS: The crude extracts of stem and leaf were subjected to phytochemicals, TPC, TTC, TFC, and free radical scavenging assays (DPPH, ABTS, etc). Excision and incision models were used to assess wound healing using the screened extracts (IPLEA, IPLM, IPSEA, and IPSM). Various tissue parameters (hydroxyproline, hexosamine, hexuronic acid content), as well as antioxidant enzyme activity (SOD, Catalase, GPX, LPO), were also examined. RESULTS: The maximum amount of polyphenolic content was found in IPLM (TPC- 118.86 ± 5.94 mg GAE/g, TTC - 75.25 ± 2.64 mg TAE/g, and TFC-25.73 ± 0.99 mg GAE/g) with significant IC50 value of 1.65 ± 0.87 µg/mL among all the extracts. Coumaric acid was reported high (92.86 mg/g) in RP-HPLC analysis of crude extract in IPLEA. The in vivo excision wound healing model revealed that 1% IPLM had better healing property with the maximum wound healing area (0.098 ± 0.03 cm) and wound concentration (95.56 ± 1.95%) was reported with the significance level of ***P < 0.001, **P < 0.01, *P < 0.05. In the incision model, IPLM represented maximum tensile strength (27500 gf). A significant functional effect of the granulation tissue parameters and enzyme antioxidants on the wound-healed area of dry tissue was also observed. Finally, the histopathological analysis showed enhanced re-epithelialization, fibroblast proliferation, and collagen synthesis in wound-treated animal tissue in both models. CONCLUSION: According to the present study, antioxidant-rich I. pes-tigridis promotes healthy cell regeneration while reducing inflammation and oxidative stress for wound healing. Additionally, it also enhances circulation and promotes healing.

Understanding the bioaccessibility, α-amylase and α-glucosidase enzyme inhibition kinetics of Allmania nodiflora (L.) R.Br. ex Wight polyphenols during in vitro simulated digestion.

This study renewed focus on Allmania nodiflora, a green leafy vegetable with diverse nutritional and medicinal properties. The bioaccessibility and the impact of in vitro simulated digestion on polyphenolics were investigated and identified using LC-MS. Although in vitro digestion reduced polyphenolics, the pancreatic digested sample showed a significant bioaccessibility of 97% with better metal ion binding activity (99%). Increased α-amylase and α-glucosidase inhibition (>45%) potentials were also observed in the digested samples. The presence of compounds such as rutin, caffeic acid, catechin, saikosaponin was also identified to be responsible for the enzyme inhibition against postprandial hyperglycemia. These results indicated that the pH of the digestive buffers is responsible for the structural changes in polyphenols for assimilation in the intestine. Hence, A. nodiflora leaf could serve as a functional food having higher assimilated polyphenolics with abundant therapeutic potential, which would be indispensible for future nutraceutical product development from green leafy vegetables.

α-Glucosidase, α-amylase inhibition kinetics, in vitro gastro-intestinal digestion, and apoptosis inducing abilities of Ficus microcarpa L. f. and Ficus racemosa L. fruit polysaccharides.

A rich source of nutrients, figs have a number of clinically validated benefits. This study aimed to evaluate the in vitro simulated gastrointestinal digestion, and the antidiabetic and anticancer activity of Ficus microcarpa (FMP) and F. racemosa (FRP) fruits polysaccharides. The pre-digested FMP revealed higher sugar content (721 ± 14.81 mg glucose equivalents/g sample) than FRP. After in vitro digestion, isolated fruit polysaccharides showed effective uptake with over 80% sugar loss. Free radicals and enzymatic inhibition after gastric digestion have been significantly modified, demonstrating the effective absorption of FMP and FRP through the intestine. Conversely, after the continuous digestion, the nutritional content of the isolated polysaccharides was gradually decreased compared to the pre-digested sample. At 30 µg/mL concentration of FMP and FRP was inhibited by 50% of breast cancer cells. The present study reveals the potential uptake of FMP and FRP as nutritional supplements in the future.