ABSTRACT
Introduction: Cerebral malaria is the most severe complication of Plasmodium falciparum infection. The pathophysiology of cerebral malaria is still unclear, but it is expected caused by cytoadherence, rosetting, autoagglutinations and abundant pro-inflammatory response that will induce the release of secondary molecules like ubiquitin, HIF-1, VEGF, and iNOS. Aim: To determine the effect of Artesunate and brotowali extract (Tinospora crispa) against the expression of ubiquitin, HIF-1α, VEGF and iNOS in the brain of cerebral malaria mice model. Methods: An experimental study of post-test only control group using CB57BL/6J mice model malaria had been done. Samples were divided into 7 groups: negative control (K-), positive control (K+), Artesunate 32 mg/BWkg/ day (P1), Tinospora crispa 70 mg/BWkg/day (P2), combination Artesunate and Tinospora crispa dose 50 mg/ kgBW (P3), combination Artesunate and Tinospora crispa dose 60 mg/kgBW (P4) and combination Artesunate and Tinospora crispa dose 70 mg/kgBW (P5). Mice model were decapitated at 7th day after infection. Expression of ubiquitin, HIF-1α, VEGF, and iNOS was measured by immunohistochemistry. Result: One Way ANOVA showed different expression of ubiquitin, HIF-1α, VEGF and iNOS among groups. Tukey test showed, there was no significant difference in expression of ubiquitin, VEGF and iNOS among single therapy (Artesunate or Tinospora crispa) with combination therapy (p>0.05). Expression of HIF-1α were significantly different between single therapy (Artesunate or Tinospora crispa) with combination therapy of Artesunate and Tinospora crispa 60 mg/kgBW (p=0.019, p=0.013) and combination therapy of Artesunate and Tinospora crispa 70 mg/kgBW (p=0.034; p=0.023). Pearson correlation showed negative correlation between Tinospora crispa dose and expression of HIF-1α (p=0.001; r=-0.832) and iNOS (p=0.001, r=-0.874). Conclusion: The combination of Artesunate and brotowali (Tinospora crispa) extract generally decreases Ubiquitin, HIF-1 α, VEGF dan iNOS expression of cerebral malaria model although only brain HIF-1α expression gives significant decreasing.
ABSTRACT
The high prevalence of pediculosis capitis, commonly known as head lice (Pediculus humanus capitis) infestation, has led to the preparation of a community-based pediculicidal ointment, which is made of common household items and the extract of Tinospora crispa stem. The present study aimed to evaluate the safety, efficacy, and physicochemical characteristics of the T. crispa pediculicidal ointment. The physicochemical properties of the ointment were characterized, and safety was determined using acute dermal irritation test (OECD 404), while the efficacy was assessed using an in vitro pediculicidal assay. Furthermore, the chemical compounds present in T. crispa were identified using liquid-liquid extraction followed by ultra-performance liquid chromatography quadruple time-of-flight mass spectrometric (UPLC-qTOF/MS) analysis. The community-based ointment formulation was light yellow in color, homogeneous, smooth, with distinct aromatic odor and pH of 6.92±0.09. It has spreadability value of 15.04±0.98 g·cm/sec and has thixotropic behavior. It was also found to be non-irritant, with a primary irritation index value of 0.15. Moreover, it was comparable to the pediculicidal activity of the positive control Kwell®, a commercially available 1% permethrin shampoo (P>0.05), and was significantly different to the activity of the negative control ointment, a mixture of palm oil and candle wax (P<0.05). These findings suggested that the community-based T. crispa pediculicidal ointment is safe and effective, having acceptable physicochemical characteristics. Its activity can be attributed to the presence of compounds moupinamide and physalin I.
Subject(s)
Chromatography, Liquid , Family Characteristics , Hydrogen-Ion Concentration , In Vitro Techniques , Lice Infestations , Liquid-Liquid Extraction , Odorants , Pediculus , Permethrin , Prevalence , TinosporaABSTRACT
To investigate the effect of Tinospora crispa (T. crispa) extract on matrix metalloproteinase 13 (MMP-13) expression and cell migration. Methods: The cytotoxicity of T. crispa extract was examined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay on head and neck squamous cell carcinoma (HNSCC) cell lines. The effect on expression of MMP-13 was analysed by RT-PCR and ELISA. The migration was assessed by wound healing assay. Results: MMP-13 mRNA was highly expressed in the metastatic human HNSCC cell lines, HN22 and HSC-3. T. crispa extract at a concentration of 100.0 μg/mL caused about 50% reduction of cell survival. T. crispa extract at a non-toxic concentration of 12.5, 25.0 and 50.0 μg/mL significantly suppressed MMP-13 mRNA expression and secreted MMP-13 in both HN22 and HSC-3. The expression of tissue inhibitors of metalloprotease by HSC-3 cells was attenuated by 25.0 and 50.0 μg/mL of T. crispa extract. Addition of the extract to cells in a wound healing assay showed inhibition of cell migration by HN22 cells. Conclusions: These data suggest that T. crispa could be considered as a potential therapeutic drug to prevent metastasis of HNSCC.
ABSTRACT
Objective: To investigate the effect of Tinospora crispa (T. crispa) extract on matrix metalloproteinase-13 (MMP-13) expression and cell migration. Methods: The cytotoxicity of T. crispa extract was examined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay on head and neck squamous cell carcinoma (HNSCC) cell lines. The effect on expression of MMP-13 was analysed by RT-PCR and ELISA. The migration was assessed by wound healing assay. Results: MMP-13 mRNA was highly expressed in the metastatic human HNSCC cell lines, HN22 and HSC-3. T. crispa extract at a concentration of 100.0 μg/mL caused about 50% reduction of cell survival. T. crispa extract at a non-toxic concentration of 12.5, 25.0 and 50.0 μg/mL significantly suppressed MMP-13 mRNA expression and secreted MMP-13 in both HN22 and HSC-3. The expression of tissue inhibitors of metalloproteinase-2 (TIMP-2) by HSC-3 cells was attenuated by 25.0 and 50.0 μg/mL of T. crispa extract. Addition of the extract to cells in a wound healing assay showed inhibition of cell migration by HN22 cells. Conclusions: These data suggest that T. crispa could be considered as a potential therapeutic drug to prevent metastasis of HNSCC.
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Background: In South Asian countries, stems of the plant, Tinnospora crispa, Linn (TC) are often used as a folk medicine in the treatment of type 2 diabetes mellitus. Though TC’s antiglycemic activity has been demonstrated in diabetic rats, the mechanism for its action has not yet been elucidated. Objective: To investigate the effect of TC’s aqueous extract (TCA) on glucose transport activity in skeletal muscle cell line. Materials and methods: A skeletal muscle cell line, L6 myoblasts, was used for this study. The myoblasts grown to the stage of fused myotubes were pre-incubated with and without TCA for 24 hours. Then, a 2-[³H]-deoxy-D-glucose (2-DG) uptake test was made in a 24-well plate for 10 minutes. In the downstream transport regulation studies, the TCA pre-incubated cells was either treated or untreated with specific inhibitors of the PI3-Kinase (wortmannin) and p38 MAP-Kinase (SB203580) pathways prior to the uptake test. All studies were carried out in triplicate with a minimum of three independent experiments. Results were expressed as mean±SE and compared with student’s t test for a level of significance at p \< 0.05. Results: TCA at 4 mg/mL significantly enhances glucose uptake of L6 myotubes in dose and time dependent manner with the half maximum effects at 24 hours (196.60±11.09%, p \< 0.05). The effect was completely abolished by a cytoskeletal blockade (10 μM of cytochalasin B), supportive of active glucose transport activity. Both wortmannin and SB203580 have no effect on the TCA-stimulated glucose uptake. Conclusion: Tinospora crispa enhances glucose transport of L6 myotubes in an insulin-independent pathway in a time- and dose-dependent manner.