Does Cleistanthin A modulate proton currents of human neutrophils in the presence of ATP?

Abirami, V (2016) Does Cleistanthin A modulate proton currents of human neutrophils in the presence of ATP? Masters thesis, Christian Medical College, Vellore.

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Abstract

Cleistanthus collinus is a poisonous plant, the leaves of which are consumed for suicide in many parts of southern India. Exact mechanism of toxicity remains unclear. Previously, two toxic principles present in this plant namely, Cleistanthin A and Cleistanthin C were tested for proton channel blockade in human neutrophils. While neither compound blocked proton channels, the impression was that Cleistanthin A may actually enhance proton currents, though the increase was not statistically significant. Since ATP was not present in the pipette in the earlier experiments, we hypothesized that in the presence of ATP, Cleistanthin A may enhance proton currents significantly. This data is important because it will help us establish this effect as the mechanism of toxicity leading to respiratory arrest in cases of Cleistanthus collinus poisoning. AIM : To study the effect of Cleistanthin A on voltage gated proton channels of human neutrophils, with ATP included in the patch pipette. OBJECTIVES : 1. To record proton currents in freshly isolated human neutrophils using Patch clamp technique. 2. To compare the percentage of proton currents remaining after the addition of the test (Cleistanthin A) and control solutions (ethanol), as compared to the currents recorded before the addition, while ATP was included in the patch pipette. METHODS : • Isolation of fresh human neutrophils. • Isolation of Cleistanthin A. • Recording proton currents in isolated neutrophils by Patch Clamp technique. • Analysis of the percentage of proton currents remaining after the addition of the test and control solutions in comparison to currents recorded before addition. RESULTS: When the proton currents recorded at different voltage-clamp levels, after the addition of either Cleistanthin A (test) or ethanol (control), were expressed as percentages of the pre-intervention currents at the same voltages, there was no statistically significant difference in the percentage current remaining after intervention in both groups. CONCLUSION: The results indicate that even in the presence of ATP, Cleistanthin A does not show any significant effect on voltage gated proton channels of human neutrophils.

Item Type: Thesis (Masters)
Uncontrolled Keywords: Cleistanthin A ; ATP ; Proton channels ; Neutrophils.
Subjects: MEDICAL > Physiology
Depositing User: Subramani R
Date Deposited: 01 Jan 2018 18:21
Last Modified: 01 Jan 2018 18:21
URI: http://repository-tnmgrmu.ac.in/id/eprint/5021

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