Quantitative Proteomics Reveal That Metabolic Improvement Contributes to the Cardioprotective Effect of T89 on Isoproterenol-Induced Cardiac Injury

Abstract

BackgroundT89, a traditional Chinese medicine, has passed phase II, and is undergoing phase III clinical trials for treatment of ischemic cardiovascular disease by the US FDA. However, the role of T89 on isoproterenol (ISO)-induced cardiac injury is unknown. The present study aimed to explore the effect and underlying mechanism of T89 on ISO-induced cardiac injury.MethodsMale Sprague-Dawley rats received subcutaneous injection of ISO saline solution at 24 h intervals for the first 3 days and then at 48 h intervals for the next 12 days. T89 at dose of 111.6 and 167.4 mg/kg was administrated by gavage for 15 consecutive days. Rat survival rate, cardiac function evaluation, morphological observation, quantitative proteomics, and Western blotting analysis were performed.ResultsT89 obviously improved ISO-induced low survival rate, attenuated ISO-evoked cardiac injury, as evidenced by myocardial blood flow, heart function, and morphology. Quantitative proteomics revealed that the cardioprotective effect of T89 relied on the regulation of metabolic pathways, including glycolipid metabolism and energy metabolism. T89 inhibited the enhancement of glycolysis, promoted fatty acid oxidation, and restored mitochondrial oxidative phosphorylation by regulating Eno1, Mcee, Bdh1, Ces1c, Apoc2, Decr1, Acaa2, Cbr4, ND2, Cox 6a, Cox17, ATP5g, and ATP5j, thus alleviated oxidative stress and energy metabolism disorder and ameliorated cardiac injury after ISO. The present study also verified that T89 significantly restrained ISO-induced increase of HSP70/HSP40 and suppressed the phosphorylation of ERK, further restored the expression of CX43, confirming the protective role of T89 in cardiac hypertrophy. Proteomics data are available via ProteomeXchange with identifier PXD024641.ConclusionT89 reduced mortality and improves outcome in the model of ISO-induced cardiac injury and the cardioprotective role of T89 is correlated with the regulation of glycolipid metabolism, recovery of mitochondrial function, and improvement of myocardial energy.

Proteomics Revealed That Mitochondrial Function Contributed to the Protective Effect of Herba Siegesbeckiae Against Cardiac Ischemia/Reperfusion Injury

“…Enzyme-linked immunosorbent assay (ELISA) was performed using a specific kit indicated by a microplate reader (RT-6100, Rayto, Shenzhen, China), respectively, to determine the content of creatine kinase-MB (CK-MB), troponin T (cTnT), lactic dehydrogenase (LDH), alanine transaminase (ALT), aspartate transaminase (AST), ROS, NO, RNS, IL-1β, IL-18, TNF-α, and IL-6 in plasma (Dogesce Biotech, Beijing, China), and 8-OHdG, malonaldehyde (MDA), and NAD + in the left ventricle (Dogesce Biotech, Beijing, China). Likewise, the activities of caspase-1, MnSOD, and mitochondrial Complexes I, II, IV, and V were also detected using the ELISA kit according to the manufacturer’s instructions ( 24 ).…”

“…Three biological replicates were analyzed, and each sample was adjusted to the same intensity of pooled internal standard within each TMT experiment. Only protein groups without any missing values in all samples were selected for Student’s t -test ( 24 ).…”

“…The specimens were processed as routine for ultrathin sections. Sections were stained with uranium acetate and lead citrate, and ultrastructural changes were then evaluated by using a transmission electron microscope (TEM) (Hitachi TEM system, Tokyo, Japan) ( 24 ).…”

Mitochondria in Pathological Cardiac Hypertrophy Research and Therapy

“…A growing number of studies have proved that phytochemicals are capable of mitigating cardiac hypertrophy in cell and animal models via modulating mitochondrial function and maintaining mitochondrial hemostasis. Wei's recent findings suggest that T89, a traditional Chinese medicine, may be a potential therapeutic approach to treat ISO-induced cardiac hypertrophy, and the underlying mechanisms mainly lay on the regulation of metabolic pathways ( 71 ). By performing quantitative proteomics, the group revealed that T 89 could inhibit glycolysis, accelerate fatty acid oxidation and restore mitochondrial OXPHOS, thereby attenuating energy metabolism disorders and alleviating ISO-induced cardiac injury.…”

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“…Experimental procedures were approved by the Animal Care Committee of Dongzhimen Hospital Affiliated to the Beijing University of Chinese Medicine (No. [21][22][23][24][25][26][27][28][29][30][31][32][33][34][35].…”