Toddalolactone Protects Against Osteoarthritis by Ameliorating Chondrocyte Inflammation and Suppressing Osteoclastogenesis

Background: Osteoarthritis (OA) is widely recognized as the most common chronic joint disease accompanied by progressive cartilage and subchondral bone damage. Toddalolactone (TOD), a natural compound extracted from Toddalia asiatica (L.) Lam., has been widely used in the treatment of stroke, rheumatoid arthritis, and oedema. Nevertheless, what TOD acts as in the pathogenesis and progression of OA hasn’t been reported. In this investigation, we have aimed to determine how TOD affects OA in vitro and in vivo.Methods: LPS (10μg/ml) was employed to induce chondrocyte inflammation or RANKL to induce osteoclast differentiation in bone marrow derived macrophages (BMMs); Meanwhile, the effects of TOD on chondrocyte inflammation and osteoclast differentiation were evaluated. Anterior cruciate ligament transection (ACLT) was performed to develop an OA animal model and study the effects of TOD.Results: We found that TOD inhibited the expression of inflammatory and catabolic mediators ( IL-6, IL-8, TNF-α, MMP2, MMP9, and MMP13) in LPS-treated chondrocytes in vitro. Furthermore, TOD was proven to inhibit RANKL-induced-osteoclastogenesis and inhibit the expression of marker genes (NFATc1 and c-fos). Our data also confirmed that TOD suppressed the destruction of articular cartilage and osteoclastogenesis by inhibiting the activation of the NF-κB and MAPK signalling pathways. In the ACLT mouse model, we found that TOD attenuated the ACLT-induced cartilage erosion and inhibited bone resorption.Conclusions: These results showed that TOD can be adopted as a potential therapeutic agent for OA.

Evaluation of Antibacterial and Antifungal Activity of Herbs Used in Treatment of Diabetes, Malaria and Pneumonia in Kisii and Nyamira Counties Region, Kenya

The herbs Carissa spinarum, Physalis minima and Toddalia asiatica have traditionally been used in healing diabetes, malaria and pneumonia by the communities around the Kisii region, Kenya. However in the available literature, there is scanty information on effectiveness of different plant parts of the herbs in healing the ailments. The objective of this study was to investigate the potential antimicrobial and antifungal activity of methanolic extract of whole plant Physalis minima, leaf and root of Carissa spinarum and Toddalia asiatica against gram positive bacteria Staphylococcus aureus (ATCC 25923), gram negative bacteria Escherichia coli (ATCC 25922) and fungus Candida albicans (ATCC14053). Antibiotic disc methicillin, cotrimoxazole, chloramphenicol, gentamicin, ampicillin, nalidixic and nitrofurantoin were used in the study. In each herb, plant part was extracted by soaking in methanol/dichloromethane in ratio 1:1 for a week, filtered, concentrated by rotary vapor and cooled. The same process was repeated three times for all samples. The study was conducted by agar well diffusion method. Methanolic root extract of Toddalia asiatica showed highest antibacterial activity against Staphylococcus aureus (ATCC 25923), root extract of Carissa spinarum had highest antibacterial activity against Escherichia coli (ATCC 25922) while root extract of Toddalia asiatica showed highest antifungal activity. It was concluded that root extract of Toddalia asiatica showed highest antibacterial activity 16.7mm against Staphylococcus aureus(ATCC 25923), root extract of Carissa spinarum had highest antibacterial activity 10 mm against Escherichia coli (ATCC 25922) while root extract of Toddalia asiatica had highest antifungal activity 18 mm against Candida albicans.

A Simple and Efficient Two-Dimensional High-Speed Counter-Current Chromatography Linear Gradient and Isocratic Elution Modes for the Preparative Separation of Coumarins from Roots of Toddalia asiatica (Linn.) Lam.

Toddalia asiatica (L.) Lam. (Rutaceae) has shown a broad spectrum of biological properties, such as anti-inflammatory, antioxidant, antimicrobial, anti-HIV, and anticancer properties. The present study is concerned with the separation of the main components with broad partition coefficients (KD values) from T. asiatica, using linear gradient high-speed counter-current chromatography (LGCCC) combined with an off-line two-dimensional (2D) mode. Similar to the binary gradient HPLC, the LGCCC mode is operated by the adjustment of the proportion between the mobile phase of 5:5:1:9 (v/v) (pump A) and 5:5:4.5:5.5 (v/v) (pump B) in an n-hexane/ethyl acetate/methanol/water solvent system. The off-line 2D-CCC mode was used in this study for the secondary separation of two similar KD value compounds with n-hexane/ethyl acetate/methanol/water (5:5:4:6, v/v). Notably, six coumarins, namely, tomentin (1), toddalolactone (2), 5,7,8-trimethoxycoumarin (3), mexoticin (4), isopimpinellin (5), and toddanone (6), were efficiently separated. The structures of the pure compounds were elucidated by spectral techniques and compared with the literature.

Pimpinellin Inhibits Collagen-induced Platelet Aggregation and Activation Through Inhibiting Granule Secretion and PI3K/Akt Pathway

Pimpinellin is a coumarin-like compound extracted from the root of Toddalia asiatica. Its effects on platelet function has not been investigated. This study found that pimpinellin pretreatment effectively inhibited collagen-induced platelet aggregation, but did not alter ADP- and thrombin-induced aggregation. Platelets pretreated with pimpinellin showed reduced α granule (CD62) level and secretion of dense granule (ATP release). Pimpinellin-treated platelets also exhibited decreased clot reaction and TxB2 production. Pimpinellin pretreatment suppressed adhesion and spreading of human platelets on the fibrinogen coated surface. Analysis of tail bleeding time of mice administered with pimpinellin (40 mg/kg) revealed that pimpinellin did not change tail bleeding time significantly, number of blood cells, and APTT and PT levels. Pimpinellin inhibited collagen-induced ex vivo aggregation of mice platelets. Immunoblotting results showed that pimpinellin suppressed collagen-induced phosphorylation of PI3K-Akt-Gsk3β and PKC/MAPK in platelets.

­­Medicinal Plant Use and Trade Among the Samburu People in Maralal Town, Kenya

Background: The Samburu people have for the longest time used traditional knowledge on medicinal plants to treat diseases both in human and livestock due to the attributed belief of how effective they are. I set out to understand medicinal plant use and trade among the Samburu community in Maralal town. The objectives of this study were to; (1) identify and document the medicinal plants traded, (2) record the medicinal uses of identified plants, (3) document the parts of the plant used and methods of preparation. Method: I conducted market surveys and held in-depth discussions with the traders on what plant parts were used and sold and where they collected and sourced their plants. I also joined collectors in the wild and collected samples for further identification. Results: From the market surveys, a total of 24 plants species from 16 families were traded. Most of the plants were collected from Kirisia Forests Reserve and the arid shrublands within Samburu County. It was noted that Myrsine africana L., Rotheca myricoides (Hochst.) Steane & Mabb., Rhamnus staddo A.Rich., Salvadora persica L. and Toddalia asiatica (L.) Lam were the most popular species with buyers. Most of the plants were used to treat multiple ailments. Stomach related problems and respiratory disorders were some of the commonly treated ailments. Conclusion: It is evident that use and trade of medicinal plant is an important economic activity in Maralal, however; overharvesting plants from the wild has contributed to some species like Osyris lanceolata Hochst. & Steud (East African sandalwood) and M. africana becoming rare and scarce hence the need to establish conservation measures and sustainable exploitation.