Vivek Kumar Gupta1, Anupam Kaushik2, Davendra Singh Chauhan3, Ramesh Kumar Ahirwar4, Shweta Sharma2, Deepa Bisht2. 1. Department of Biochemistry, National JALMA Institute for Leprosy & Other Mycobacterial Diseases (ICMR), Tajganj, Agra 282004, Uttar Pradesh, India. Electronic address: gupta.vk2@icmr.gov.in. 2. Department of Biochemistry, National JALMA Institute for Leprosy & Other Mycobacterial Diseases (ICMR), Tajganj, Agra 282004, Uttar Pradesh, India. 3. Department of Microbiology and Molecular Biology, National JALMA Institute for Leprosy & Other Mycobacterial Diseases (ICMR), Tajganj, Agra 282004, Uttar Pradesh, India. 4. Department of Botany, Govt. College Birsinghpur Pali, Umaria 484551, Madhya Pradesh, India.
Abstract
ETHNOPHARMACOLOGICAL RELEVANCE: Tuberculosis (TB) is one of the highly infectious disease caused by the bacterium Mycobacterium tuberculosis (Mtb). Several medicinal plants are traditionally used by the tribal healers of some tribal pockets of Madhya Pradesh (M.P.), India in the treatment of various ailments including TB related symptoms. The information of traditional knowledge of plants is empirical lacking systematic scientific investigation. AIM OF THE STUDY: To provide a scientific rationale for the traditional uses of some medicinal plants in treating prolonged cough, chest complaints and TB by ethnic groups in four districts of the state of M.P., India. The anti-mycobacterial activity of these plants was also investigated against multidrug-resistant (MDR) strains of Mtb. The active plant extracts were also assessed for general cytotoxicity by human monocytic leukemia cell line, THP-1 macrophages. MATERIALS AND METHODS: Semi structured interviews and guided field-walk methods were used to gather information on medicinal plants used by the tribal traditional healers. The study was carried out in the tribal pockets of Anuppur, Mandla, Umariya and Dindori districts of M.P., India. A total of 35 plant species distributed in 22 families were selected based on folklore uses suggested by the tribal traditional healer. The ethanol extracts of plants were evaluated for anti-mycobacterial activity against Mtb H37Rv and six MDR clinical isolates of Mtb. The anti-mycobacterial activity of plants was determined in terms of minimum inhibitory concentration (MIC) using standard resazurin microtitre plate assay (REMA). The plant extracts found to be active against Mtb were further evaluated for general cytotoxicity against human THP-1 macrophages using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium (MTT) assay. The selectivity index (SI) of active plant extracts were also calculated on the basis of MIC and cytotoxicity. RESULTS: Out of the 35 plants, 11 plant species showed anti-mycobacterial activity with MIC ranging from 500 to 31.25 µg/mL against Mtb H37Rv. The plant extracts also exhibited anti-mycobacterial activity against six multi-drug resistant (MDR) clinical strains of Mtb isolated from sputum samples of pulmonary TB patients. In vitro cytotoxicity of active plant extracts was also assessed in human THP-1 macrophages. The IC50 (50% inhibitory concentration) values of most of the plant extracts on THP-1 was found to be higher than MIC values against Mtb, indicating that the THP-1 cells are not adversely affected at concentrations that are effective against Mtb. Significant anti-mycobacterial activity was observed for eleven plants viz., Alstonia scholaris (L.) R. Br., Glycyrrhiza glabra L., Holorrhena antidysentrica (Roth) Wall. exA.DC., Mallotus philippensis (Lam.) Müll. Arg., Eulophia nuda Lindl., Cocculus hirsutus (L.) Diels, Pueraria tuberosa (Willd.) DC., Cyperus rotundus L., Curcuma caesia Roxb., Sphaeranthus indicus L. and Plumbago zeylanica L. which lends support to their traditional uses. CONCLUSION: The present investigation supports the potential role of plants used by tribal healers as our results have shown that these plants exhibit anti-mycobacterial activity in the acceptable range against Mtb. Our study clearly lends support to the traditional uses of some plants in TB related symptoms as we have found them to exhibit significant in vitro anti-TB activity. To the best of our knowledge, anti-mycobacterial activity in plants against Mtb is being reported for the first time from tribal areas of M.P., India by the current study.
ETHNOPHARMACOLOGICAL RELEVANCE: Tuberculosis (TB) is one of the highly infectious disease caused by the bacterium Mycobacterium tuberculosis (Mtb). Several medicinal plants are traditionally used by the tribal healers of some tribal pockets of Madhya Pradesh (M.P.), India in the treatment of various ailments including TB related symptoms. The information of traditional knowledge of plants is empirical lacking systematic scientific investigation. AIM OF THE STUDY: To provide a scientific rationale for the traditional uses of some medicinal plants in treating prolonged cough, chest complaints and TB by ethnic groups in four districts of the state of M.P., India. The anti-mycobacterial activity of these plants was also investigated against multidrug-resistant (MDR) strains of Mtb. The active plant extracts were also assessed for general cytotoxicity by human monocytic leukemia cell line, THP-1 macrophages. MATERIALS AND METHODS: Semi structured interviews and guided field-walk methods were used to gather information on medicinal plants used by the tribal traditional healers. The study was carried out in the tribal pockets of Anuppur, Mandla, Umariya and Dindori districts of M.P., India. A total of 35 plant species distributed in 22 families were selected based on folklore uses suggested by the tribal traditional healer. The ethanol extracts of plants were evaluated for anti-mycobacterial activity against MtbH37Rv and six MDR clinical isolates of Mtb. The anti-mycobacterial activity of plants was determined in terms of minimum inhibitory concentration (MIC) using standard resazurin microtitre plate assay (REMA). The plant extracts found to be active against Mtb were further evaluated for general cytotoxicity against humanTHP-1 macrophages using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium (MTT) assay. The selectivity index (SI) of active plant extracts were also calculated on the basis of MIC and cytotoxicity. RESULTS: Out of the 35 plants, 11 plant species showed anti-mycobacterial activity with MIC ranging from 500 to 31.25 µg/mL against MtbH37Rv. The plant extracts also exhibited anti-mycobacterial activity against six multi-drug resistant (MDR) clinical strains of Mtb isolated from sputum samples of pulmonary TBpatients. In vitro cytotoxicity of active plant extracts was also assessed in humanTHP-1 macrophages. The IC50 (50% inhibitory concentration) values of most of the plant extracts on THP-1 was found to be higher than MIC values against Mtb, indicating that the THP-1 cells are not adversely affected at concentrations that are effective against Mtb. Significant anti-mycobacterial activity was observed for eleven plants viz., Alstonia scholaris (L.) R. Br., Glycyrrhiza glabra L., Holorrhena antidysentrica (Roth) Wall. exA.DC., Mallotus philippensis (Lam.) Müll. Arg., Eulophia nuda Lindl., Cocculus hirsutus (L.) Diels, Pueraria tuberosa (Willd.) DC., Cyperus rotundus L., Curcuma caesia Roxb., Sphaeranthus indicus L. and Plumbago zeylanica L. which lends support to their traditional uses. CONCLUSION: The present investigation supports the potential role of plants used by tribal healers as our results have shown that these plants exhibit anti-mycobacterial activity in the acceptable range against Mtb. Our study clearly lends support to the traditional uses of some plants in TB related symptoms as we have found them to exhibit significant in vitro anti-TB activity. To the best of our knowledge, anti-mycobacterial activity in plants against Mtb is being reported for the first time from tribal areas of M.P., India by the current study.
Authors: Amir Roointan; Reza Kamali-Kakhki; Mohammad Fathalipour; Zohreh Hashemi; Mohammad Mehdi Zarshenas; Mohammad Soleimani; Ruhola Mirjani Journal: Iran J Med Sci Date: 2020-11