The nature of cell death and mechanism associated with it is yet to be reported. Thus in this part of this thesis several investigations were attempted to provide possible mechanism of the nature and mode of cell death seen with a selected panel of kratom high point nc woolwine human cell lines. Kratom In Capsules Dosage the cytological examination using three different cell lines (SH-SY5Y HEK 293 and MCL-5 cells) was the first investigation. As anticipated toxicity Kratom In Capsules Dosage effects seen at high doses suggested apoptotic morphology with evidence of chromatin condensation which was predominantly seen in SH-SY5Y cells. Nuclear alterations are key in many descriptions of apoptosis. The severity of MSE insult in the SH-SY5Y cell line was obvious at the highest dose tested as there were very few cells present on the slide and all of them showed apoptotic morphology. For HEK 293 cells the nature of cell death was more necrotic than Kratom In Capsules Dosage apoptotic as morphologically the cell membrane integrity was compromised leaving a reduced stained intensity and indicating lysis of
cell membrane and subsequent lost of cell content.
Y Y Y Y Y Y Y Y Y Y Y Conc. Summary table of MLA result for MSE in the i) presence of S9 and ii) in the absence of S9. S9 treatment Treatment groups Negative control MSE 0 0 0 40 30 20 Positive control (MMS) Mean Control MF 75. Negative Negative Negative Negative Negative Negative Positive Conc.
MSE and MIT. From these estimates it appears that the SH-SY5Y cells are the most sensitive of those examined to the cytotoxic and possibly cytostatic effect of MSE. Based upon my estimation of 42% MIT-like compound in MSE extract the SHSY5Y cell IC50 for MSE is equal to what does kratom smell like 9.
Prominent apoptotic-like cell death is mainly observed for SH-SY5Y cells and a necrotic type of cell death for the MCL-5 and HEK-293 cells. Further confirmation on these findings in differentiating the Kratom In Capsules Dosage stages of cell death was carried out using Annexin V conjugate assay via flow cytometry analysis with SH-SY5Y and MCL-5 cells. Unfortunately difficulties in interpreting the analysis were encountered as dose-dependant shifts in dye uptake were found as in the earlier cell cycle analysis. The right shifting of the whole cell population made the interpretation of apoptotic and necrotic populations very difficult as they were not located in the anticipated quadrants thus the results remain inconclusive. This finding however gives strong justification to the hypothesised mechanism discussed earlier in which MSE and MIT may have the ability to change membrane permeabilisation or cause pore opening.
MSE and 2. M MIT respectively (Table 2. M -5 3. D ) in MSE and MIT treated HEK 293 cells as determined by the Trypan blue exclusion assay. SH-SY5Y cells With SH-SY5Y cells low doses MSE (0. These higher doses of MSE also substantially increased cell death within 24 hr (Fig.
The two most abundant oxindoles are mitraphylline and speciofoline. Other alkaloids present include ajmalicine corynanthedine mitraversine rhychophylline and stipulatine. Mitragynine is believed by many to be but has not been proven to be the primary active alkaloid in M. The effects of kratom can be described as comparable to opium based-products but milder. In general the effects are stimulating and euphoric at a lower doses and are more calming and narcotic at higher doses. These effects are noticeable after 5 to 10 minutes and can last for several hours. Kratom contains a number of active components so-called alkaloids of which mitragynine is believed to be responsible for most of its effects.
For cytotoxicity assay; MSE treated kratom effects length HepG2 cells were cultured as described in section 2. C for 10 min. The reaction was terminated with stop solutions provided with the kit. The plate was read using a best strain kratom euphoria eden valley fluorescent plate reader with an excitation wavelength of 560 nm and emission wavelength of 590 nm.