6th World Congress of Oxidative Stress, Calcium Signalling and TRP Channels, Isparta, Turkey, 24 - 27 May 2016, vol.8, no.1, pp.497
Lignosus rhinocerostis (cooke) Prevents Neurotoxic Effects of Cisplatin on Cultured Sensory Neurons
Ramazan Üstün1, Elif Kaval Oğuz2, Ayşe Şeker1, Sıddık Keskin3
1Yuzuncu Yıl University, Faculty of Medicine, Department of Physiology, Van, Turkey
2Yuzuncu Yıl University, Faculty of Education, Division of Science Teaching, Van, Turkey
3Yuzuncu Yıl University, Faculty of Medicine, Department of Biostatistics, Van, Turkey
Cisplatin is effective chemotherapy drug in cancer treatment, induces peripheral neuropathy in 30% of patients. Peripheral neuropathy is the dose limiting side effect, which has no preventative therapy.
The purpose of this study is to examine the regenerative effect of Lignosus rhinocerostis on cultured sensory neurons exposed to the toxic effect of cisplatin.
Sensory neurons are grown in primary culture following enzymatic and mechanical dissociation of dorsal root ganglia from 6-8-week-old mice. Neuron culture are incubated for 24 hours under the moist condition which has 5% CO2 circulation at 37 °C. Propidium iodure is added to culture to visualize dead cells and Calcein AM (1μM) is added to visualize the survival cells.
1) Cisplatin: Neurons are exposed for 24 hours with cisplatin (10μg/ml).
2) Lignosus r. extract + cisplatin: Neurons are treated for 1 h with lignosus r. extract (30 μg/ml) and then exposed for 24 hours with cisplatin (10μg/ml).
3) K252a + Lignosus r. extract + cisplatin: Firstly neurons are treated for 1 h with k252a (inhibitor of trkA) (100nM) and then treated for 1 h with lignosus r. extract (30 μg/ml) and finally exposed for 24 hours with cisplatin (10μg/ml)
4) Control: No application is made
Microscopic visualization: “Zeiss Cell Observer”, a microscopic visualization system is used. Mozaix mode is chosen for visualizing the survival and the dead neuron cells and changes in the neuritis. Two proportions Z test is used for comparison of groups’ proportions for survival rates of neurons, survival rates of neuritis and rates of fragmentation. In addition, Wilcoxon test is also used to compare before and after values of axon length.
At the end of the study, the survival rates of neurons in the groups (cisplatin, lignosus, k252a, control) are observed as 275/313, 204/206, 274/277 and 265/269, respectively. The survival rates of neuritis are also recorded as 90/261, 193/206, 272/277, 265/269, respectively. The rates of fragmentation in neuritis are found 67/261, 8/206, 5/277, 1/269 in the groups.
There is no statistically significant differences among the lignosus, k252a and control groups in terms of rates of survival neuron, neurite and fragmentation as well as axon length. However, cisplatin group is found different from the other three groups (p < 0,01).
As a conclusion, it can be stated that Lignosus rhinocerostis (cooke) prevents neurotoxic effects of cisplatin on cultured sensory neurons and it has therapeutic potential.
Key words: Lignosus, Cisplatin, Dorsal root ganglion neuron