Objectives: Isatis tinctoria L., clary, is an herbal plant traditionally used in folk medicine for the treatment of various diseases and conditions. Although it has been primarily used as an antimicrobial and antifung...Objectives: Isatis tinctoria L., clary, is an herbal plant traditionally used in folk medicine for the treatment of various diseases and conditions. Although it has been primarily used as an antimicrobial and antifungal, there are data on traditional use of I. tinctoria as an agent against antiallergic, anti-thrombocytosis. The aim of the study was to examine the effect of the I. tinctoria root extract on the lipopolysaccharide (LPS)-induced periodontitis in rats on osteoclast associated bone resorptive activity, cell death including apoptosis, and inflammation in a rat of disease model of periodontitis. Materials and Methods: Periodontitis, acute or chronic inflammatory status in periodontal tissue in rats could be induced by repeated injections of LPS from Escherichia coli into the periodontal pocket area between the first and second right maxillary molars. Eighteen male rats were distributed among the following treatment groups: 1) I. tinctorial root extract (Antifect) 200 mg/kg body weight, 2) acetylsalicylic acid (ASA), 20 mg/kg body weight and 3) Phosphate buffered saline (PBS) treatment used as a control. After 15 days, maxilla, alveolar bone, molar teeth and associated periodontal tissues were harvested. Inflammatory alveolar bone resorption was analyzed by microcomputerized tomography (μCT) (microcomputer tomography). Tissues fixed with paraformaldehyde and formalin for 2 days, after that paraffin embedded histological sections were stained with haematoxylin and eosin (H/E) for the assessment of histopathological changes or tested to immunohistochemistry for detecting TRAP (tartrate resistant acid phosphatase) positive cells and caspase 3. Cell death and Apoptosis were analyzed in the periodontal tissues by tunnel assay. The inflammatory status was assessed by the measurements of proinflammatory cytokines interleukin-Iβ (IL-Iβ), interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) of gingival tissues and descriptive analysis of histological sections of periodontal. Results: Treatment with Antifect, compared to the control group, significantly diminished the process of inflammation decreasing the levels of IL-Iβ, IL-6 and TNF-α, reducing the gingival tissue lesions and preserving bone alveolar resorption. Considerably a smaller number of inflammatory cells and a larger number of fibroblasts were noticed. Also, μCT analysis showed that only Antifect treated group reduced bone resorption and the number of TRAP-positive multinucleated cells (osteoclasts), also, significantly reduced the number of apoptotic cells in the gingival tissues and of osteocytes in the alveolar bone crest. Conclusion: Antifect manifested anti-inflammatory elect and reducing alveolar bone resorption in LPS-induced periodontitis suggest that it may have a role as a therapeutic agent in periodontal diseases.展开更多
Traditional Chinese medicine(TCM)belongs to the most elaborate and extensive systems of plant-based healing.The herb Northern Ban Lan(Isatis tinctoria)is famous for its antiviral and anti-inflammatory activity.Althoug...Traditional Chinese medicine(TCM)belongs to the most elaborate and extensive systems of plant-based healing.The herb Northern Ban Lan(Isatis tinctoria)is famous for its antiviral and anti-inflammatory activity.Although numerous components isolated from I.tinctoria have been characterized so far,their modes of action have remained unclear.Here,we show that extracts from I.tinctoria exert antimicrotubular activity.Using time-lapse microscopy in living tobacco BY-2(Nicotiana tabacum L.cvBrightYellow2)cellsexpressing greenfluorescentprotein-tubulin,weuse activity-guided fractionation to screen out the biologically active compounds of I.tinctoria.Among 54 fractions obtained from either leaves or roots of I.tinctoria by methanol(MeOH/H_(2)O 8:2),or ethyl acetate extraction,one specific methanolic root fraction was selected,because it efficiently and rapidly eliminated microtubules.By combination of further purification with ultrahigh-performance liquid chromatography and high-resolution tandem mass spectrometry most of the bioactivity could be assigned to the glucosinolate compound glucobrassicin.Glucobrassicin can also affect microtubules and induce apoptosis in HeLa cells.In the light of these findings,the antiviral activity of Northern Ban Lan is discussed in the context of microtubules being hijacked by many viral pathogens for cell-to-cell spread.展开更多
文摘Objectives: Isatis tinctoria L., clary, is an herbal plant traditionally used in folk medicine for the treatment of various diseases and conditions. Although it has been primarily used as an antimicrobial and antifungal, there are data on traditional use of I. tinctoria as an agent against antiallergic, anti-thrombocytosis. The aim of the study was to examine the effect of the I. tinctoria root extract on the lipopolysaccharide (LPS)-induced periodontitis in rats on osteoclast associated bone resorptive activity, cell death including apoptosis, and inflammation in a rat of disease model of periodontitis. Materials and Methods: Periodontitis, acute or chronic inflammatory status in periodontal tissue in rats could be induced by repeated injections of LPS from Escherichia coli into the periodontal pocket area between the first and second right maxillary molars. Eighteen male rats were distributed among the following treatment groups: 1) I. tinctorial root extract (Antifect) 200 mg/kg body weight, 2) acetylsalicylic acid (ASA), 20 mg/kg body weight and 3) Phosphate buffered saline (PBS) treatment used as a control. After 15 days, maxilla, alveolar bone, molar teeth and associated periodontal tissues were harvested. Inflammatory alveolar bone resorption was analyzed by microcomputerized tomography (μCT) (microcomputer tomography). Tissues fixed with paraformaldehyde and formalin for 2 days, after that paraffin embedded histological sections were stained with haematoxylin and eosin (H/E) for the assessment of histopathological changes or tested to immunohistochemistry for detecting TRAP (tartrate resistant acid phosphatase) positive cells and caspase 3. Cell death and Apoptosis were analyzed in the periodontal tissues by tunnel assay. The inflammatory status was assessed by the measurements of proinflammatory cytokines interleukin-Iβ (IL-Iβ), interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) of gingival tissues and descriptive analysis of histological sections of periodontal. Results: Treatment with Antifect, compared to the control group, significantly diminished the process of inflammation decreasing the levels of IL-Iβ, IL-6 and TNF-α, reducing the gingival tissue lesions and preserving bone alveolar resorption. Considerably a smaller number of inflammatory cells and a larger number of fibroblasts were noticed. Also, μCT analysis showed that only Antifect treated group reduced bone resorption and the number of TRAP-positive multinucleated cells (osteoclasts), also, significantly reduced the number of apoptotic cells in the gingival tissues and of osteocytes in the alveolar bone crest. Conclusion: Antifect manifested anti-inflammatory elect and reducing alveolar bone resorption in LPS-induced periodontitis suggest that it may have a role as a therapeutic agent in periodontal diseases.
基金supported by fellowships from the China Scholarship Council。
文摘Traditional Chinese medicine(TCM)belongs to the most elaborate and extensive systems of plant-based healing.The herb Northern Ban Lan(Isatis tinctoria)is famous for its antiviral and anti-inflammatory activity.Although numerous components isolated from I.tinctoria have been characterized so far,their modes of action have remained unclear.Here,we show that extracts from I.tinctoria exert antimicrotubular activity.Using time-lapse microscopy in living tobacco BY-2(Nicotiana tabacum L.cvBrightYellow2)cellsexpressing greenfluorescentprotein-tubulin,weuse activity-guided fractionation to screen out the biologically active compounds of I.tinctoria.Among 54 fractions obtained from either leaves or roots of I.tinctoria by methanol(MeOH/H_(2)O 8:2),or ethyl acetate extraction,one specific methanolic root fraction was selected,because it efficiently and rapidly eliminated microtubules.By combination of further purification with ultrahigh-performance liquid chromatography and high-resolution tandem mass spectrometry most of the bioactivity could be assigned to the glucosinolate compound glucobrassicin.Glucobrassicin can also affect microtubules and induce apoptosis in HeLa cells.In the light of these findings,the antiviral activity of Northern Ban Lan is discussed in the context of microtubules being hijacked by many viral pathogens for cell-to-cell spread.