Assessment of biological activity and UPLC-MS based chromatographic profiling of ethanolic extract of Ochradenus arabicus

Abstract

Natural products from wild and medicinal plants, either in the form of crude extracts or pure compounds provide unlimited opportunities for new drug leads owing to the unmatched availability of chemical diversity. In the present study, the cytotoxic potential of crude ethanolic extract of Ochradenus arabicus was analyzed by MTT cell viability assay in MCF-7 adenocarcinoma breast cancer cells. We further investigated its effect against oxidative stress induced by anticancer drug doxorubicin. In addition, Ultra Performance Liquid Chromatography-Mass Spectrometry (UPLC-MS) based chromatographic profiling of crude extract of O. arabicus was performed. The MTT assay data showed that the extract is moderately toxic to the MCF-7 cells. However, its treatment alone does not induce oxidative stress while doxorubicin increases the level of oxidative stress in MCF-7 cells. Whereas, simultaneous treatment of plant extract and doxorubicin significantly (p < 0.05) decreased the level of intracellular reactive oxygen species (ROS) and lipid peroxidation while an increase in the reduced glutathione and superoxide dismutase activity was observed in time and dose dependent manner. Hence, our finding confirmed cytotoxic and antioxidant potential of crude extract of O. arabicus in MCF-7 cells. However, further investigations on O. arabicus as a potential chemotherapeutic agent are needed. The analysis of bioactive compounds present in the plant extracts involving the applications of common phytochemical screening assays such as chromatographic techniques is discussed.

Keywords: Cytotoxicity; Ochradenus arabicus; Oxidative stress; ROS; UPLC–MS.

Figure 1

Inhibition of MCF-7 cell proliferation by crude ethanolic extract of Ochradenus arabicus. Cells were treated with indicated concentrations of extract for 24 h, and cell viability was determined by the MTT assay.

Figure 2

Inhibition of MCF-7 cell proliferation by crude ethanolic extract of Ochradenus arabicus. Cells were treated with indicated concentrations of extract for 24 h, and cell viability was determined by the MTT assay. The IC₅₀ value was estimated at 562 μg/ml (indicated by arrow).

Figure 3

Percentage of DCF fluorescence for detection of ROS in MCF-7 cells after exposure of Ochradenus arabicus extract and doxorubicin either alone or in combination for 12 h and 24 h. Each value represents the mean ± SE of three experiments (P < 0.05). ^∗indicates significant from Control, ^#indicates significant protective effects from doxorubicin.

Figure 4

Levels of lipid peroxides in MCF-7 cells after exposure of Ochradenus arabicus extract and doxorubicin either alone or in combination for 12 h and 24 h. Each value represents the mean ± SE of three experiments (P < 0.05). ^∗indicates significant from Control, ^#indicates significant protective effects from doxorubicin.

Figure 5

Depletion in glutathione levels in MCF-7 cells after exposure of Ochradenus arabicus extract and doxorubicin either alone or in combination for 12 h and 24 h. Each value represents the mean ± SE of three experiments (P < 0.05). ^∗indicates significant from Control, ^#indicates significant protective effects from doxorubicin.

Figure 6

Levels of SOD activity in MCF-7 cells after exposure of Ochradenus arabicus extract and doxorubicin either alone or in combination for 12 h and 24 h. Each value represents the mean ± SE of three experiments (P < 0.05). ^∗indicates significant from Control, ^#indicates significant protective effects from doxorubicin.

Figure 7

UPLC–MS spectrum obtained by screening of ethanolic extract of Ochradenus arabicus in −Ve and +Ve ESI mode.