LC-MS characterized methanolic extract of zanthoxylum armatum possess anti-breast cancer activity through Nrf2-Keap1 pathway: An in-silico, in-vitro and in-vivo evaluation
Authors: Salona Bharti
Priyashree Sunita
Roja Sahu
Sharad Srivastava
Rajiv Kumar Kar
Shakti P Pattanayak
Puja Kumari
Pritha Bose
Journal: Journal of Ethnopharmacology (2021)
Discipline: PHARMACOLOGY & TOXICOLOGY
Keywords: Antioxidant; Breast cancer; Kelch-like ECH-Associated protein 1; LC-MS/MS; Nuclear factor erythroid 2
Affiliations: Division of Pharmacology, Department of Pharmaceutical Sciences and Technology, Birla Institute of Technology, Mesra, Ranchi, 835 215, Jharkhand, India., Fritz Haber Center for Molecular Dynamics, Institute of Chemistry, The Hebrew University of Jerusalem, Jerusalem, Israel., Government Pharmacy Institute, Department of Health, Family Welfare and Medical Education, Government of Jharkhand, Bariatu, Ranchi, 834009, India., Pharmacognosy & Ethnopharmacology Division, CSIR-National Botanical Research Institute, NBRI-Govt. of India, Lucknow, 226001, India., Division of Pharmacology, Department of Pharmaceutical Sciences and Technology, Birla Institute of Technology, Mesra, Ranchi, 835 215, Jharkhand, India; Department of Pharmacy, School of Health Sciences, Central University of South Bihar (Gaya), Bihar, 824236, India. Electronic address: .

Abstract:  Ethnopharmacological relevance: Zanthoxylum armatum DC (Rutaceae) containing flavonoids, alkaloids, coumarins, lignans, amides and terpenoid is well-known for its curative properties against various ailments including cancer. In the current research, phytochemicals present in the methanolic extract of Zanthoxylum armatum bark (MeZb) were characterized by LC-MS/MS analysis and chemotherapeutic potential of this extract was determined on DMBA-induced female Sprague Dawley rats. Materials and methods: A simple and fast high-performance liquid chromatography-mass spectroscopy (LC-MS/MS) of MeZb was established followed by in-vitro antioxidant assays. This was followed by in-silico docking analysis as well as cytotoxicity assessment. Successively in-vivo study of MeZb was performed in DMBA-induced Sprague Dawley rats possessing breast cancer along with detailed molecular biology studies involving immunofluorescence, RT-qPCR and Western blot analysis. Results: LC-MS/MS investigation revealed the presence of compounds belonging to flavonoid, alkaloid and glycoside groups. MeZb revealed potential antioxidant activity in in-vitro antioxidant assays and strong binding energy of identified compounds was seen from the in-silico study with both HO1 and Keap1 receptor. Furthermore, the antioxidant action of MeZb was proven from the in-vivo analysis of antioxidant marker enzymes (lipid peroxidation, enzymic and non-enzymic antioxidants). This study also revealed upregulation of protective Nrf-2 following downregulation of Keap1 after MeZb treatment with respect to untreated cancerous rats. Conclusion: These results exhibited anti-breast-cancer potential of MeZb through Nrf2-Keap1 pathway which may be due to the flavonoids, alkaloids and glycosides present in it. 

Retracted: No

PubPeer comments:
 Figures 7D and 7E appear to share the same source, albeit with different dimensions. *** ![file] *** ![file] , 
 #1 Also Figures 7A and 7F appear to share the same source (cyan boxes), although (A) is control and (F) is Glibenclamide (10 mg kg) treatment. Could the authors please comment? ![file] , 
 Several micrographs in this paper were reused from another paper published by the same group a year before in very same journal, Kumar et al. 2013 BMC Compl Altern Med: https: pubpeer.com publications 26665E7D8B2E873C329648C9FF7A32 , 
 #3 In Figure 11 three panels (colored boxes) were published a year before in yet another paper by same group, **Kumar et al. 2013 SpringerPlus** in its Figure 22, but conditions are different. https: pubpeer.com publications 248668BE11C7245FAF6C8CEC1628BA#1 Figure 11: Paederia Foetida Linn. leaves extract effect on pancreas histopathology in normal and STZ induced diabetic rats. Figure 22: Effect of UFD photomicrographs of histological changes in rat pancreas. Could the authors please comment? EDIT: in Figure 11, panels A and F are also identical ![file] , 
 #3 This is what I meant with comment #3, treatments are different between papers, image reuse sometimes occurs also within same figure: ![file] , 
 Figure 4, several images were published a year before in [*Kumar et al. 2013 BMC Compl. Altern. Med. 13:222*](http: pubpeer.com publications DC5A54E9A031300BD9E2C3ECBEC2CE) by the same group, where the compound analyzed was different. ![file] ,