Berberine Inhibits Oxygen Consumption Rate Independent of Alteration in Cardiolipin Levels in H9c2 Cells

Abstract

Small clinical studies have shown that oral treatment with the plant alkaloid berberine (BBR) reduces blood glucose levels similar to that of metformin and have promoted its use as a novel anti‐diabetic therapy. However, in vitro studies have shown that high concentrations of BBR potently inhibit cell proliferation through inhibition of mitochondrial function. Cardiolipin (Ptd2Gro) is a key phospholipid required for regulating mitochondrial bioenergetic function. We examined if BBR inhibited oxygen consumption rate in H9c2 cardiac myocytes through alteration in Ptd2Gro metabolism. Treatment of H9c2 cells with BBR resulted in a rapid (within minutes) concentration‐dependent decrease in the oxygen consumption rate (OCR) as determined using a Seahorse XF24 analyzer. Concentrations of BBR as low as 1 µM were effective in inhibiting OCR. In addition, all concentrations of BBR inhibited the fatty acid‐mediated increase in OCR that was observed in untreated cells. Treatment of H9c2 cells with up to 25 µM BBR for 24 h markedly reduced [3H]thymidine incorporation into cells but did not alter the pool size of Ptd2Gro. In contrast, 12.5 µM BBR increased [1‐14C]palmitate incorporation into Ptd2Gro and 12.5 µM and 25 µM BBR reduced [1‐14C]oleate incorporation into Ptd2Gro. Protein kinase C delta (PKCδ) activation through its increased membrane association is known to alter Ptd2Gro distribution within mitochondria. BBR treatment resulted in a decrease in membrane‐associated PKCδ and attenuated the palmitate‐mediated increase in PKCδ membrane‐association. Thus, BBR treatment of H9c2 cardiac myocytes inhibits cellular OCR independent of alteration in Ptd2Gro levels.

Berberine for prevention of dementia associated with diabetes and its comorbidities: A systematic review

“…In addition, berberine protected cardiomyocytes from HG-induced damage and ischemia-reperfusion via enhancement of AMPK [116,120,124,132], PPARα [115] and PI3K-Akt-eNOS anti-apoptotic signaling pathways [116]. Berberine also attenuated cardiac fibrosis, possibly by reducing insulinlike growth factor (IGF)-1 receptor expression in cardiac fibroblasts [129], and enhanced lipid metabolism in cardiomyocytes, possibly via protein kinase C inhibition [119,127]; it also suppressed cardiac arrhythmia by modulating K + and Ca 2+ [110,113]. Furthermore, berberine may alleviate vascular dysfunction in diabetes by acting on adipose tissues.…”

Amorphous solid dispersion of Berberine mitigates apoptosis via iPLA2β/Cardiolipin/Opa1 pathway in db/db mice and in Palmitate-treated MIN6 β-cells

“…In addition, we found that BBR could enhance the expression of iPLA 2 β in MIN6 mitochondria. Chang et al have observed that BBR can increase CL levels in H9C2 cells 19. Given these results, we hypothesized that BBR could inhibit beta-cell apoptosis by enhancing the iPLA 2 β/CL/Opa1 pathway.…”

Reduced protein kinase C delta association with a higher molecular weight complex in mitochondria of Barth Syndrome lymphoblasts

“…For Blue-Native polyacrylamide gel electrophoresis (BN-PAGE) analysis, mitochondrial protein (80 µg) was treated with 0.2% digitonin and then separated on a 3-12% gradient gel as described [9]. Immunoblot analysis of the gel was performed using anti-PKCδ antibody (1:1000) (Abcam, Toronto, ON, Canada) as described [17]. PKCδ was visualized using the Amersham Enhanced Chemiluminescence Western blotting detection system (VWR, was not certified by peer review) is the author/funder.…”

“…Immunoblot analysis of the gel was performed using anti-PKCδ antibody (1:1000) (Abcam, Toronto, ON, Canada) as described [17]. PKCδ was visualized using the Amersham Enhanced Chemiluminescence Western blotting detection system (VWR, Mississauga, ON, Canada).…”