Anti-inflammatory and anti-arthritic effects of 3-hydroxy, 2-methoxy sodium butanoate from the leaves of Clerodendrum phlomidis L.f.

INTRODUCTION: The leaves of Clerodendrum phlomidis L.f. have been used in the Indian traditional system of medicine to treat several inflammatory diseases and arthritis. The aim of the present study was to assess the anti-inflammatory and anti-arthritic activities of the leaves of C. phlomidis and to isolate the active principle by bioactivity guided fractionation. MATERIALS AND METHODS: To find the anti-inflammatory constituents from this plant, fractionations were performed with concurrent bioassays. Carrageenan-induced inflammation and Freund complete adjuvant (FCA)-induced arthritic rat models were used. The anti-inflammatory and anti-arthritic activities of the isolated compound were studied by assessing the histology of the joints, levels of lysosomal enzymes, protein-bound carbohydrates, acute phase protein, etc., in plasma, as well as by estimating the levels and expression of pro-inflammatory cytokines in the joints. RESULTS: Repeated fractionations and bioassays yielded a novel bioactive compound: 3-hydroxy, 2-methoxy-sodium butanoate. Treatment with this compound reduced the paw edema induced by carrageenan and FCA dose dependently. The levels of lysosomal enzymes and protein-bound carbohydrates decreased significantly upon treatment with the compound. The level of plasma acute phase protein was also decreased compared with control animals. Protein levels and mRNA expression of pro-inflammatory cytokines TNF, IL-1 and IL-6 in the joints were decreased significantly in a dose-dependent manner and the histopathological data also added evidence of the anti-arthritic property of the compound. CONCLUSION: The 3-hydroxy,2-methoxy sodium butanoate isolated from plant leaves displays considerable potency in anti-inflammatory action and has a prominent anti-arthritic effect. This is the first report of this natural compound with bioactivity.

Ameliorative effect of Drynaria quercifolia (L.) J. Sm., an ethnomedicinal plant, in arthritic animals.

Drynaria quercifolia (L.) J. Sm., is an ethnomedicinal plant used widely in Tamil Nadu to treat arthritis. The present study was aimed to evaluate the traditional claim of D. quercifolia rhizome water extract in adjuvant induced arthritic animals. Anti-arthritic effect was studied by assessing the levels of lysosomal enzymes, protein bound carbohydrates, urinary degradative collagen and serum cytokines on control and adjuvant induced arthritis. The paw swelling and body weight were also analyzed. The levels of ROS and lysosomal enzymes in neutrophils of control and adjuvant induced animals were also estimated. D. quercifolia rhizome water extract at doses of 100 and 200mg/kg reduced the paw thickness and elevated the mean body weight of arthritic rats. The treatment with extract showed a significant reduction in the levels of plasma and liver lysosomal enzymes as well as protein bound carbohydrates and urinary degradative collagen levels. The treatment reduced the levels of ROS and lysosomal enzymes in neutrophils significantly. The significant reduction in the levels of serum pro-inflammatory cytokines (TNF-α and IL-1ß) and the increment in the levels of anti-inflammatory cytokine (IL-10) were also observed by the treatment. The present study supports the traditional claim of using D. quercifolia to treat rheumatism.

Immunomodulatory potential of Enicostema axillare (Lam.) A. Raynal, a traditional medicinal plant.

ETHNOPHARMACOLOGICAL RELEVANCE: Enicostema axillare (Lam.) A. Raynal., (Gentianaceae) has been used in traditional Indian system of medicine as depurative and for the treatment of psoriasis, intermittent fever and cancer. Ethnobotanical survey conducted in Theni District indicated a high consensus for this plant as blood purifier and to treat dermatopathy and venereal infections. The present study evaluated the immunomodulatory activity of the methanol extract of Enicostema axillare. MATERIALS AND METHODS: In vivo immunomodulatory activity of Enicostema axillare methanol extract (100 and 200mg/kg b.w) was evaluated by assessing its effect on the total and differential leukocyte count, organ weight, hemaggultinating antibody titer, plaque forming cells, quantitative hemolysis of SRBC and delayed type hypersensitivity. Sheep red blood cells (SRBC, 5×10(8)cells/0.1mL) were used to immunize the animals. In vitro immunomodulatory potential of the extract was studied using peritoneal macrophages by evaluating its effects on NBT reduction, NO production and cytokine release. RESULTS: The animals treated with Enicostema axillare methanol extract showed a significant (P≤0.05) increase in weight of the thymus and spleen. The total leukocyte and lymphocyte count was increased significantly (P<0.005) by the treatment. There was no significant alteration in neutrophil count. A dose dependent increase in antibody titer value was observed. A decreased response to DTH reaction induced by SRBC was recorded. A potential phagocytic response was seen on treatment with the extracts at 10 and 25µg/mL. The extract inhibited the release of pro-inflammatory cytokines and production of NO significantly in a dose dependent manner. CONCLUSION: These findings suggested that the methanol extract of Enicostema axillare acted on both humoral and cell mediated immune functions and decreased the release of pro-inflammatory cytokines in the peritoneal macrophages.

Antimicrobial activity of Ulopterol isolated from Toddalia asiatica (L.) Lam.: a traditional medicinal plant.

ETHNOPHARMACOLOGICAL RELEVANCE: The leaves of Toddalia asiatica (L.) Lam. (Rutaceae) are widely used in folk medicine in India to treat various ailments like cough, malaria, indigestion, influenza lung diseases and rheumatism, fever, stomach ailments, cholera and diarrhea. In our earlier communication we have reported the antimicrobial study on the various extracts of the leaves and the isolation and identification of Flindersine, a quinolone alkaloid as the major active principle. In the present study, we report the antibacterial and antifungal activities of Ulopterol, a coumarin isolated as another major active antimicrobial principle. MATERIALS AND METHODS: The leaves were successively extracted with hexane, chloroform, ethyl acetate, methanol and water. The extracts were studied for their antimicrobial activity against selected bacteria and fungi by using disc-diffusion method. The ethyl acetate extract which was found to possess highest antimicrobial activity was subjected to activity guided fractionation by column chromatography over silica gel. This resulted in the isolation of the coumarin, Ulopetrol, an active principle besides Flindersine which was reported by us earlier. The structure of the compound was elucidated using physical and spectroscopic data. Flindersine and Ulopterol were quantified by HPLC. RESULTS: Ulopterol showed activity against the bacteria viz. Staphylococcus epidermidis, Enterobacter aerogenes, Shigella flexneri, Klebsiella pneumoniae (ESBL-3967), Escherichia coli (ESBL-3984) and fungi viz. Aspergillus flavus, Candida krusei and Botrytis cinerea. Quantification by HPLC showed the content of Flindersine and Ulopterol to be 0.361% and 0.266% respectively on dry weight basis of the leaves. CONCLUSIONS: Ethyl acetate extract (successive extraction) contained Ulopterol, a coumarin, besides Flindersine, a quinolone alkaloid, as a major active principle in the antimicrobial studies. This is the first report of the antimicrobial activity of Ulopterol and also its first report from the plant.