Anthelmintic, anticancer and phytochemical screening of Cotyledon orbiculata; Hermannia depressa; Nicotiana glauca and potassium permanganate
Molefe, Nthatisi Innocentia
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Cotyledon orbiculata, Hermannia depressa and Nicotiana glauca are widely used plants in traditional medicine for treatment of various infections and diseases. C. orbiculata is mostly used in treatment of epilepsy, earache and the removal of warts while H. depressa is used to relieve toothache, nerves and diarrhoea. Lastly, N. glauca has been mostly used as pain killer in relieving earache, toothache and seldom applied on body in treatment of soreness and inflammation. Traditional usages of these plants by indigenous people and local traditional healers have not yet been scientifically reported, that is why the current study was aimed at determining the anthelmintic and anticancer activity of these plants. The anthelmintic activity of acetone and water extracts from the shoots of Cotyledon orbiculata, Hermannia depressa and Nicotiana glauca were investigated using the egg hatch, larval development and larval mortality assays. In all extracts tested, C. orbiculata water extract at 7.5 mg/ml prevented nematode eggs from hatching with 82.63% success rate. Other extracts exhibited egg hatch inhibition in a degree of less than 50%. Similarly, C. orbiculata water extracts suppressed nematode larval development with 85.32% at the concentration of 2.5 mg/ml followed by 66.69% of H. depressa extract at 7.5 mg/ml concentration. However, N. glauca water and all acetone extracts induced the 100% larval development inhibition. The in vitro larval mortality rate revealed that the water extracts from all the plants were able to kill all larvae at 2.5 mg/ml within 48 to 96 h. The results from this study have shown that the extracts from the three plants have the potential to prevent and ameliorate diseases associated with gastrointestinal nematodes. The anticancer activity of the above mentioned plants was tested on two cancer cell lines, the MCF-7 and HeLa cells. All plants possessed anticancer activity at different durations and concentrations. The N. glauca demonstrated an activity against both cell lines, however, the plant acetone extracts were much effective on the MCF-7 line at 48 hours with inhibition percentages > 80% at all concentrations. The N. glauca acetone extracts were effective at 24 hours. The H. depressa acetone extracts also possessed much activity than water extracts at 24 hours, whereas C. orbiculata showed no activity at all on this cell line. The C. orbiculata acetone extracts followed by the water extracts were the most effective on the HeLa line ranging from 12 to 84% and 0 to 77% inhibition. H. depressa activity was observed at 48 hours of experimentation at all concentrations in both extracts. N. glauca exhibited significant inhibition percentages at 24 hours of water extracts and 48 hours of acetone extracts. This study has showed that the three plants are potential candidates for cancer treatment. Brine shrimp lethality test was conducted on the nauplii stage of the shrimps. Furthermore, two assays, MTT and LDH cytotoxicity assays were conducted on the MDBK cells. None of the plants was toxic on the brine shrimps as most of them survived through 24 and 48 hours. C. orbiculata and H. depressa brine shrimp larval mortality was observed at 96, 72, 72, 96 and 96 h at 0.2, 0.4, 0.6, 0.8, and 1.0 mg/ml of water extracts respectively and 72 h at all concentrations of acetone extracts while N. glauca larval mortality was observed at 120 and 96 hours at all concentration of both extracts. There was a cell survival decrease of 50% at 0.65 mg/ml and 50-60% in C. orbiculata water and acetone extracts treated wells, respectively. No significant cell decrease was observed from both H. depressa and N. glauca water and acetone extracts except for N. glauca acetone extract at 2.50 mg/ml. None of the plants induced a significant toxicity on both the brine shrimps and the bovine cells. Alkaloids, saponins, tannins and phenols were the constituents tested for in order to identify constituents responsible for observed activity of the whole study. No alkaloids were detected from all plants for both extracts, only saponins, tannins and phenols were present even though N. glauca acetone extracts possessed none of the tested constituents. Tested plants are therefore good candidates in treating parasitic gastrointestinal nematodes and cancer disease. Potassium permanganate (KMnO₄) is a widely used chemical substance in the rural communities as prophylaxis or in treatment of various infections and diseases; however, there is no scientific validation of its usage. Anthelmintic activity of potassium permanganate was tested against parasitic gastrointestinal nematodes in comparison to anthelmintic commercial drugs, Tramisol®, Noromectin® and Valbazen®. There was no egg hatch inhibition induced (<10%) by potassium permanganate at all concentrations as compared to >80% inhibition of commercial drugs, however, 85.24, 98.10 and 90.91% of larval development was induced at 0.5, 1.0 and 1.5 mg/ml, respectively. Larval mortality was observed at 216, 144 and 144 hours at 0.5, 1.0 and 1.5 mg/ml, respectively. The cytotoxicity of this chemical was tested on the brine shrimps and MBDK cells as well. Within 24 hours all the brine shrimps larvae treated with KMnO₄ died. A significant decrease in MDBK cell survival was observed at 0.65 mg/ml at 24, 48 and 72 hours. This study has therefore showed that potassium permanganate have anthelmintic activity and concentration dependent cytotoxicity on the mammalian cell lines. In conclusion Cotyledon orbiculata, Hermannia depressa, Nicotiana glauca and potassium permanganate have the potential to supress the aggressiveness of parasitic gastrointestinal nematode and cancer diseases. These substances are therefore potential candidates in treatment of tested pathogens.