JBM  Vol.7 No.9 , September 2019
Anti-Inflammatory Effect of the Isatis tinctoria L. Root Extract on Lipopolysaccharide-Induced Periodontitis in Rats
Abstract: 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.
Cite this paper: Lee, S. , Cho, Y. (2019) Anti-Inflammatory Effect of the Isatis tinctoria L. Root Extract on Lipopolysaccharide-Induced Periodontitis in Rats. Journal of Biosciences and Medicines, 7, 81-95. doi: 10.4236/jbm.2019.79008.

[1]   Kolenbrander, P.E. (1993) Coaggregation of Human Oral Bacteria: Potential Role in the Accretion of Dental Plaque. Journal of Applied Bacteriology, 74, 79S-86S.

[2]   Oliver, D.R. (1968) The Etiology of Periodontal Disease. The Journal of the Missouri Dental Association, 48, 12-18.

[3]   Lang, N.P. (2014) Commentary: Bacteria Play a Critical Role in the Etiology of Periodontal Disease. Journal of Periodontology, 85, 211-213.

[4]   Golz, L., et al. (2014) LPS from P. gingivalis and Hypoxia Increases Oxidative Stress in Periodontal Ligament Fibroblasts and Contributes to Periodontitis. Mediators of Inflammation, 2014, Article ID: 986264.

[5]   Stein, S.H., Borke, J.L. and Cummings, L.A. (1999) Effects of Chronic Adult Periodontitis and Endotoxin (LPS) on Gingival Fibroblast Plasma Membrane Ca -Pump. Connective Tissue Research, 40, 59-66.

[6]   Sharma, N., et al. (2014) Cytokine Gene Polymorphism (Interleukin-1Beta +3954, Interleukin-6 [-597/-174] and Tumor Necrosis Factor-Alpha -308) in Chronic Periodontitis with and without Type 2 Diabetes Mellitus. Indian Journal of Dental Research, 25, 375-380.

[7]   Teles, F.R., Teles, R.P., Martin, L., Socransky, S.S. and Haffajee, A.D. (2012) Relationships among Interleukin-6, Tumor Necrosis Factor-Alpha, Adipokines, Vitamin D, and Chronic Periodontitis. Journal of Periodontology, 83, 1183-1191.

[8]   Babel, N., et al. (2006) Analysis of Tumor Necrosis Factor-Alpha, Transforming Growth Factor-Beta, Interleukin-10, IL-6, and Interferon-Gamma Gene Polymorphisms in Patients with Chronic Periodontitis. Journal of Periodontology, 77, 1978-1983.

[9]   Kanzaki, H., et al. (2017) Pathways that Regulate ROS Scavenging Enzymes, and Their Role in Defense against Tissue Destruction in Periodontitis. Frontiers in Physiology, 8, 351.

[10]   Chiang, C.Y., et al. (2016) A Salvia miltiorrhiza Ethanol Extract Ameliorates Tissue Destruction Caused by Experimental Periodontitis in Rats. Journal of Periodontal Research, 51, 133-139.

[11]   Yoshinaga, Y., et al. (2014) Green Tea Extract Inhibits the Onset of Periodontal Destruction in Rat Experimental Periodontitis. Journal of Periodontal Research, 49, 652-659.

[12]   Barrella, G.E., et al. (2012) Evaluation of the Effect of an Organic Extract Obtained from Ipomoea alba L. on Experimental Periodontitis in Rats. Brazilian Oral Research, 26, 158-164.

[13]   Anand, P., Kunnumakkara, A.B., Newman, R.A. and Aggarwal, B.B. (2007) Bioavailability of Curcumin: Problems and Promises. Molecular Pharmacology, 4, 807-818.

[14]   Liou, C.J., Huang, W.C., Kuo, M.L., Yang, R.C. and Shen, J.J. (2010) Long-Term Oral Administration of Gynostemma pentaphyllum Extract Attenuates Airway Inflammation and Th2 Cell Activities in Ovalbumin-Sensitized Mice. Food and Chemical Toxicology, 48, 2592-2598.

[15]   Kerdar, T., Rabienejad, N., Alikhani, Y., Moradkhani, S. and Dastan, D. (2019) Clinical, in Vitro and Phytochemical, Studies of Scrophularia striata Mouthwash on Chronic Periodontitis Disease. Journal of Ethnopharmacology, 239, 111872.

[16]   Heinemann, C., Schliemann-Willers, S., Oberthür, C., Hamburger, M. and Elsner, P. (2004) Prevention of Experimentally Induced Irritant Contact Dermatitis by Extracts of Isatis tinctoria Compared to Pure Tryptanthrin and Its Impact on UVB-Induced Erythema. Planta Medica, 70, 385-390.

[17]   Danz, H., Baumann, D. and Hamburger, M. (2002) Quantitative Determination of the Dual COX-2/5-LOX In-hibitor Tryptanthrin in Isatis tinctoria by ESI-LC-MS. Planta Medica, 68, 152-157.

[18]   Seifert, K. and Unger, W. (1994) Insecticidal and Fungicidal Compounds from Isatis tinctoria. Zeitschrift für Naturforschung C, 49, 44-48.

[19]   Danz, H., Stoyanova, S., Wippich, P., Brattström, A. and Hamburger, M. (2001) Identification and Isolation of the Cyclooxygenase-2 Inhibitory Principle in Isatis tinctoria. Planta Medica, 67, 411-416.

[20]   Zhang, D., et al. (2019) Indole Alkaloid Glycosides from Isatis tinctoria Roots. Natural Product Research, 1-7.

[21]   Zhou, J. and Qu, F. (2011) Analysis of the Extracts of Isatis tinctoria by New Analytical Approaches of HPLC, MS and NMR. African Journal of Traditional, Complementary and Alternative Medicines, 8, 33-45.

[22]   Brattstrom, A., et al. (2010) The Plant Extract Isatis tinctoria L. Extract (ITE) Inhibits Allergen-Induced Airway Inflammation and Hyperreactivity in Mice. Phytomedicine, 17, 551-556.

[23]   Dumitrescu, A.L., Abd El-Aleem, S., Morales-Aza, B. and Donaldson, L.F. (2004) A Model of Periodontitis in the Rat: Effect of Lipopolysaccharide on Bone Resorption, Osteoclast Activity, and Local Peptidergic Innervation. Journal of Clinical Periodontology, 31, 596-603.

[24]   Zhang, Z.S. (2018) Association between COX2 -765G/C Polymorphism and Periodontitis in Chinese Population: A Meta-Analysis. BMC Oral Health, 18, 32.

[25]   Kamon, M., Fujita, D., Goto, N., Amano, H. and Sakamoto, K. (2008) Prostaglandin F2α Negatively Regulates Bone Resorption in Murine Osteoclast Development. Prostaglandins & Other Lipid Mediators, 87, 26-33.

[26]   Acharya, A.B., Thakur, S., Muddapur, M.V. and Kulkarni, R.D. (2016) Tumor Necrosis Factor-Alpha, Interleukin-4 and -6 in the Serum of Health, Chronic Periodontitis, and Type 2 Diabetes Mellitus. Journal of Indian Society of Periodontology, 20, 509-513.

[27]   Martin, P.L., et al. (2005) Reviews Preclinical Safety and Immune-Modulating Effects of Therapeutic Monoclonal Antibodies to Interleukin-6 and Tumor Necrosis Factor-alpha in Cynomolgus Macaques. Journal of Immunotoxicology, 1, 131-139.

[28]   Fujikawa, Y., Sabokbar, A., Neale, S.D., Itonaga, I., Torisu, T. and Athanasou, N.A. (2001) The Effect of Macrophage-Colony Stimulating Factor and Other Humoral Factors (Interleukin-1, -3, -6, and -11, Tumor Necrosis Factor-Alpha, and Granulocyte Macrophage-Colony Stimulating Factor) on Human Osteoclast Formation from Circulating Cells. Bone, 28, 261-267.

[29]   Stashenko, P., Jandinski, J.J., Fujiyoshi, P., Rynar, J. and Socransky, S.S. (1991) Tissue Levels of Bone Resorptive Cytokines in Periodontal Disease. Journal of Periodontology, 62, 504-509.

[30]   Recio, M.C., Cerdá-Nicolás, M., Potterat, O., Hamburger, M. and Ríos, J.-L. (2006) Anti-Inflammatory and Antiallergic Activity in Vivo of Lipophilic Isatis tinctoria Extracts and Tryptanthrin. Planta Medica, 72, 539-546.

[31]   Gao, B., Zhang, J. and Xie, L. (2018) Structure Analysis of Effective Chemical Compounds against Dengue Viruses Isolated from Isatis tinctoria. Canadian Journal of Infectious Diseases and Medical Microbiology, 2018, Article ID: 3217473.

[32]   Ahmad, I. and Fatima, I. (2008) Butyrylcholinesterase, Lipoxygenase Inhibiting and Antifungal Alkaloids from Isatis tinctoria. Journal of Enzyme Inhibition and Medicinal Chemistry, 23, 313-316.