Effect of Cleistanthin A on Voltage Gated Proton Channels of Human Neutrophils

Introduction: Cleistanthus collinus (C. collinus), a well known plant toxin, contains active principles like Cleistanthin A, Cleistanthin B, Cleistanthin C and Diphyllin. Previous human case reports and animal studies have revealed that C. collinus poisoning leads to type I Distal renal tubular acid...

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Main Authors: Rajasegaran Rajalakshmi, Soosai Manickam Amirtham, V Abirami, Sathya Subramani, Praghalathan Kanthakumar
Format: Article
Language:English
Published: JCDR Research and Publications Private Limited 2018-01-01
Series:Journal of Clinical and Diagnostic Research
Subjects:
Online Access:https://jcdr.net/articles/PDF/11124/31729_F(SHU)_PF1_(BT_PB_RA_SL)_PFA(GG_MJ)_PN(GG).pdf
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author Rajasegaran Rajalakshmi
Soosai Manickam Amirtham
V Abirami
Sathya Subramani
Praghalathan Kanthakumar
author_facet Rajasegaran Rajalakshmi
Soosai Manickam Amirtham
V Abirami
Sathya Subramani
Praghalathan Kanthakumar
author_sort Rajasegaran Rajalakshmi
collection DOAJ
description Introduction: Cleistanthus collinus (C. collinus), a well known plant toxin, contains active principles like Cleistanthin A, Cleistanthin B, Cleistanthin C and Diphyllin. Previous human case reports and animal studies have revealed that C. collinus poisoning leads to type I Distal renal tubular acidosis and type II respiratory failure. However, the mechanism of toxicity of this plant is still uncertain. Based on the hypothesis that blockade of proton channels could result in type II respiratory failure, patch clamp experiments were done to see if Cleistanthin A blocked the proton channels. Aim: To record and compare the changes in the magnitude of voltage-gated proton currents in human neutrophils, before and after addition of Cleistanthin A (test) and control solution. Materials and Methods: The test compound Cleistanthin A was isolated by partition chromatography and characterised using thin layer chromatography. Neutrophils were isolated by density gradient centrifugation method. Using voltage clamp protocol, proton currents were recorded before (pre-intervention currents) and after (post-intervention currents) the addition of Cleistanthin A or control solution. The pre and post-intervention current densities for different voltages were compared within the groups (control and test) by Wilcoxon signed-rank test and the percentage current remaining in both the groups were compared using Mann-Whitney U test, p<0.05 was considered significant. Results: Normal proton currents were recorded in human neutrophils. Comparison of the pre and post-intervention current densities within the control and test group revealed a significant depletion effect in the control group but not in the test group. However, comparison of the percentage current remaining after intervention across the groups did not show any significant difference between the control and test groups. Conclusion: Cleistanthin A does not seem to have any significant effect on the voltage-gated proton channels of human neutrophils.
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spelling doaj.art-16ecb532c6d4483284750b1a9f9396fc2022-12-22T01:24:04ZengJCDR Research and Publications Private LimitedJournal of Clinical and Diagnostic Research2249-782X0973-709X2018-01-01121CC05CC0810.7860/JCDR/2018/31729.11124Effect of Cleistanthin A on Voltage Gated Proton Channels of Human NeutrophilsRajasegaran Rajalakshmi0Soosai Manickam Amirtham1V Abirami2Sathya Subramani3Praghalathan Kanthakumar4Assistant Professor, Department of Physiology, Indira Gandhi Medical College and Research Institute, Kathirkamam, Puducherry, India.Associate Research Officer, Department of Physiology, Christian Medical College, Vellore, Tamil Nadu, India.Assistant Professor, Department of Physiology, Thiruvarur Medical College, Thiruvarur, Tamil Nadu, India.Professor and Head, Department of Physiology, Christian Medical College, Vellore, Tamil Nadu, India.Assistant Professor, Department of Physiology, Christian Medical College, Vellore, Tamil Nadu, India.Introduction: Cleistanthus collinus (C. collinus), a well known plant toxin, contains active principles like Cleistanthin A, Cleistanthin B, Cleistanthin C and Diphyllin. Previous human case reports and animal studies have revealed that C. collinus poisoning leads to type I Distal renal tubular acidosis and type II respiratory failure. However, the mechanism of toxicity of this plant is still uncertain. Based on the hypothesis that blockade of proton channels could result in type II respiratory failure, patch clamp experiments were done to see if Cleistanthin A blocked the proton channels. Aim: To record and compare the changes in the magnitude of voltage-gated proton currents in human neutrophils, before and after addition of Cleistanthin A (test) and control solution. Materials and Methods: The test compound Cleistanthin A was isolated by partition chromatography and characterised using thin layer chromatography. Neutrophils were isolated by density gradient centrifugation method. Using voltage clamp protocol, proton currents were recorded before (pre-intervention currents) and after (post-intervention currents) the addition of Cleistanthin A or control solution. The pre and post-intervention current densities for different voltages were compared within the groups (control and test) by Wilcoxon signed-rank test and the percentage current remaining in both the groups were compared using Mann-Whitney U test, p<0.05 was considered significant. Results: Normal proton currents were recorded in human neutrophils. Comparison of the pre and post-intervention current densities within the control and test group revealed a significant depletion effect in the control group but not in the test group. However, comparison of the percentage current remaining after intervention across the groups did not show any significant difference between the control and test groups. Conclusion: Cleistanthin A does not seem to have any significant effect on the voltage-gated proton channels of human neutrophils.https://jcdr.net/articles/PDF/11124/31729_F(SHU)_PF1_(BT_PB_RA_SL)_PFA(GG_MJ)_PN(GG).pdfchromatographycleistanthus collinuspatch clampproton currents
spellingShingle Rajasegaran Rajalakshmi
Soosai Manickam Amirtham
V Abirami
Sathya Subramani
Praghalathan Kanthakumar
Effect of Cleistanthin A on Voltage Gated Proton Channels of Human Neutrophils
Journal of Clinical and Diagnostic Research
chromatography
cleistanthus collinus
patch clamp
proton currents
title Effect of Cleistanthin A on Voltage Gated Proton Channels of Human Neutrophils
title_full Effect of Cleistanthin A on Voltage Gated Proton Channels of Human Neutrophils
title_fullStr Effect of Cleistanthin A on Voltage Gated Proton Channels of Human Neutrophils
title_full_unstemmed Effect of Cleistanthin A on Voltage Gated Proton Channels of Human Neutrophils
title_short Effect of Cleistanthin A on Voltage Gated Proton Channels of Human Neutrophils
title_sort effect of cleistanthin a on voltage gated proton channels of human neutrophils
topic chromatography
cleistanthus collinus
patch clamp
proton currents
url https://jcdr.net/articles/PDF/11124/31729_F(SHU)_PF1_(BT_PB_RA_SL)_PFA(GG_MJ)_PN(GG).pdf
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