IC50 values (Inhibition concentration that caused 50% cell death) of 24 hr treatment with MSE and MIT treated cell lines. The values were interpolated from percentage dead cells curves obtained from the Trypan blue exclusion experiments. Captain Kratom Thai Capsules Review Doran mIT (Molar) 7. 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.
In modern times people from cultures around the globe have incorporated the powder into comprehensive approaches to kratom legal bali yukon well-being. But as every plant interacts slightly differently with every user sometimes a more potent variation is desirable. For this purpose the technique of extraction was created. This dark gummy substance dries into a smooth hard rock which can then be crushed and ground up easily. It is highly concentrated with a rating indicating the ratio of Captain Kratom Thai Capsules Review Doran original leaves Captain Kratom Thai Capsules Review Doran to final product.
However the potential cytotoxicity of this plant is unknown. Therefore the cytotoxicity of methanol-chloroform extract (MSE) and MIT on human cell lines (HepG2 HEK 293 MCL-5 Captain Kratom Thai Capsules Review Doran cHol and SH-SY5Y cells) has been examined. SH-SY5Y was the most sensitive cell line examined. MIT showed a similar response. Clonogenicity assay was performed to assess the longer- term effects of MSE and MIT. The colony forming ability of HEK 293 and SH-SY5Y cells was inhibited in a dose-dependant manner. Involvement of metabolism in cytotoxicity was further assessed by clonogenicity assay using Captain Kratom Thai Capsules Review Doran rat liver S9 (induced by Arochlor 1254); toxicity increased 10-fold in both cell lines.
To further clarify the above finding S9 from rat liver (induced by Arochlor 1254) was used with SH-SY5Y and HEK-293 cells as these cells have no metabolic what is the best strain of kratom for pain activity. MSE in SHSY5Y and HEK 293 cells respectively; this cytotoxic dose of MSE is ten fold lower than with cells treated without S9. CYP 1A2 inhibitor) and 3-amino 124triazole (CYP 2E1 inhibitor) were used with MCL-5 cells and analysed for cytotoxicity. MIT toxicity was not possible. Introduction The results from trypan
blue exclusion experiments and clonogenicity assays described in the previous chapter (chapter 2) demonstrated that MSE and MIT were cytotoxic in the cell lines examined.
After 3 hr incubation the cells were washed with PBS (for SH-SY5Y cells) or D-PBS (for HEK 293 cells) by centrifugation resuspended in drug-free medium and reseeded for clonogenicity as described above. To further examine the involvement of metabolism in MSE and MIT associated toxicity specific inhibitors of metabolic enzymes were used. M ketoconazole (KT) a CYP 3A4 inhibitor (Gibbs et al. M 3-amino-124-triazole (ATZ) a CYP2E1 inhibitor (Koop 1990).
Absorbance 227 nm 2 1. Calibration curve for MIT. M under standard conditions of room temperature.
This assessment can either be tailored to determine cell morphology characteristics biochemical or even the molecular changes. Various methods have been developed for identification of living and dead cells which could easily be differentiated during microscopic examinations or by other means such as fluorescence using a plate reader or by flow cytometry analysis. The methods developed were based on difference capability of intracellular intake or dye processing between live and dead cells:
- Biochemical assessments confirmed that MSE induced cell death independent of p53 or caspases pathway while MIT cell death appeared to be associated with p53 and caspases pathway
- M checkpoints (Pellegata et al 1996)
- Studies with opioid antagonists were performed using SH-SY5Y cells treated with MSE and MIT
- Mitragyna speciosa extracts
. Such methods includes the use of coloured dyes such as trypan blue eosin nigrosin or fast green or fluorescence dyes such as fluoresceine diacetate propidium iodide acridine orange or ethidium bromide (Cianco et al 1988). As discussed in section 1. The use of common histochemistry staining such as Wright-Giemsa stain which contains methylene blue and eosin will aid in identifying the nucleus and cytoplasm based on different colouration methylene blue stained nucleus blue-purplish and eosin stained cytoplasm pink (Colomick et al 1979). Microscopic mitragyna speciosa bali south solon technique may also be used to study the detailed morphology of cell death (apoptosis) define mitragyna speciosa by using electron microscopy (Odaka and Ucker 1996).