Comparison of toad skins Bufo bufo gargarizans Cantor from different regions for their active constituents content and cytotoxic activity on lung carcinoma cell lines

Abstract

Background:The skin of Bufo bufo gargarizans Cantor, rich in bufadienolides, peptides, and alkaloids, has approved pharmacological activity for preliminary anti-liver and lung tumor treatment. However, few studies have systematically focused on the influence of the producing regions on the content and antitumor activity of the active constituents in toad skins.Objective:This study aims to compare toad skins obtained from six different regions in China (Jiangsu, Anhui, Henan, Hebei, Jiangxi, and Shandong province) for their bufadienolide and alkaloid content, and their cytotoxic activity on two lung carcinoma cell lines (SPC-A-1 cells and A549 cells).Materials and Methods:High performance liquid chromatography (HPLC) was used to quantificationally determine four bufadienolides, which included bufotalin, bufalin, cinobufagin, and resibufogenin in toad skins, from six different regions, respectively. In addition, an ultraviolet (UV) spectrophotometer was also employed to identify the content of the total alkaloids using 5-hydroxytryptamine (5-HT) as the reference substance. An MTT assay was performed to compare the antiproliferative effects of the toad skins’ ethanolic extracts from the different regions against SPC-A-1 and A549 cells.Results:In this study, the toad skins from Jiangsu province had the highest amount of bufadienolides (472.6 μg/g crude drug) and alkaloids (1.51 mg/g crude drug). Meanwhile, according to the extract, it exhibited the strongest cytotoxic effect against the lung carcinoma cell line (SPC-A-1 cells and A549 cells) with IC50 values of 24.82 ± 0.76 and 23.77 ± 0.63 μg crude drug/mL, respectively.Conclusion:The toad skins that originated from the Jiangsu province, have comparatively greater advantages over samples from other regions as far as active constituent content and potential anti-lung cancer activity is concerned, suggesting that it can be a promising chemotherapeutic agent in lung cancer therapy, in further studies.

The spinal microglial IL-10/β-endorphin pathway accounts for cinobufagin-induced mechanical antiallodynia in bone cancer pain following activation of α7-nicotinic acetylcholine receptors

“…Being listed in the Chinese Pharmacopeia, Bufonis venenum has been used as a therapeutic agent in Asian countries for centuries to treat cardiovascular diseases, inflammatory diseases, cancers and chronic pain [12,70,80,87,88,91]. The major active components in Bufonis venenum are bufadienolides, such as cinobufagin, bufalin, resibufogenin, cinobufotalin, bufotenine, bufotenidine, and bufobutanoicacid, the first three of which account for approximately 10% of the dry weight of toad venom [43,46,47,84,86]. The antiinflammatory and antinociceptive effects of Bufonis venenum and its purified bufadienolides have been extensively studied over the past years.…”

“…Cinobufagin is probably the most abundant bufadienolide in Bufonis venenum and has displayed significant analgesic effect [35,43,46,47,84,86]. Therefore, it was selected in this study to evaluate the antinociceptive effects of Bufonis venenum in cancer pain and explore the underlying mechanisms.…”

Toxins and pharmacologically active compounds from species of the family Bufonidae (Amphibia, Anura)

“…Owing to the well-known usage of bufonids in medicinal remedies and the proven pharmacological potential offered by the great chemical diversity of these steroids, the occurrence among species has been summarized (Table S-3). This summary table meets the need for a more complete chemical profile among species of Bufonidae (Gao et al, 2010b;Liu et al, 2014;Zhao et al, 2014).…”

Arenobufagin activates p53 to trigger esophageal squamous cell carcinoma cell apoptosis in vitro and in vivo

“…In traditional Chinese medicine, amphibian skin extract has been used for the treatment of different diseases. Previous studies have demonstrated that the major pharmacological constituents derived from toad skins are hydrosoluble indole alkaloids (bufotenine, bufotenidine, and cinobufotenine) and liposoluble steroidal cardiac glycosides mainly composed of bufadienolides 3,4. Bufadienolide-type cardiotonic steroids are a class of C-24 steroids with a characteristic pyrone ring at C-17 and have been identified as major active components and marker compounds of venenum bufonis 5.…”