Mechanism Study of the Protective Effects of Sodium Tanshinone IIA Sulfonate Against Atorvastatin-Induced Cerebral Hemorrhage in Zebrafish: Transcriptome Analysis

Abstract

Hemorrhage stroke is a severe vascular disease of the brain with a high mortality rate in humans. Salvia miltiorrhiza Bunge (Danshen) is a well-known Chinese Materia Medica for treating cerebral vascular and cardiovascular diseases in traditional Chinese medicine. Sodium tanshinone IIA sulfonate (STS) is a water-soluble derivative of tanshinone IIA, which is the main active ingredient of Danshen. In our previous study, we established a zebrafish model of cerebral hemorrhage and found that STS dramatically decreased both the hemorrhage rate and hemorrhage area, although the underlying mechanism was not fully elucidated. We conducted a transcriptome analysis of the protective effect of STS against atorvastatin (Ator)-induced cerebral hemorrhage in zebrafish using RNA-seq technology. RNA-seq revealed 207 DEGs between the Ator-treated group and control group; the expression levels of 53 DEGs between the Ator-treated group and control group were reversed between the STS + Ator-treated group and Ator-treated group. GO enrichment analysis indicated that these 53 DEGs encode proteins with roles in hemoglobin complexes, oxygen carrier activity and oxygen binding, etc. KEGG analysis suggested that these 53 DEGs were most enriched in three items, namely, porphyrin and chlorophyll metabolism, ferroptosis, and the HIF-1 signaling pathway. The PPI network analysis identified 12 hub genes, and we further verified that Ator elevated the mRNA expression levels of hemoglobin ( hbae1.3 , hbae3 , hbae5, hbbe2 , and hbbe3 ), carbonic anhydrase ( cahz) , HIF-1 ( hif1al2 ) and Na+/H+ exchanger ( slc4a1a and slc9a1 ) genes, while STS significantly suppressed these genes. In addition, we found that pharmacological inhibition of PI3K/Akt, MAPKs, and mTOR signaling pathways by specific inhibitors partially attenuated the protective effect of STS against Ator-induced cerebral hemorrhage in zebrafish, regardless of mTOR inhibition. We concluded that hemoglobin, carbonic anhydrase, Na+/H+ exchanger and HIF-1 genes might be potential biomarkers of Ator-induced cerebral hemorrhage in zebrafish, as well as pharmacological targets of STS. Moreover, HIF-1 and its regulators, i.e., the PI3K/Akt and MAPK signaling pathways, were involved in the protective effect of STS against Ator-induced cerebral hemorrhage. This study also provided evidence of biomarkers involved in hemorrhage stroke and improved understanding of the effects of HMG-COA reductase inhibition on vascular permeability and cerebral hemorrhage.

Isosteviol Sodium Exerts Anti-Colitic Effects on BALB/c Mice with Dextran Sodium Sulfate-Induced Colitis Through Metabolic Reprogramming and Immune Response Modulation

“…Yu et al reported similar findings 55 and found that IBD is associated to changes in lipid metabolism and energy metabolic intermediates that occur to match the metabolic demands of intestinal inflammation. Hemoglobin is dependent on chlorophyll and porphyrin metabolisms, 56 while riboflavin metabolism may be related to the production of inflammatory cytokines in IBDs. 57 STV-Na administration significantly changed these metabolic pathways by inhibiting inflammation.…”

Luteolin Prevents Cardiac Dysfunction and Improves the Chemotherapeutic Efficacy of Doxorubicin in Breast Cancer

“…Tg (cmlc2: GFP) zebrafish with GFP specifically expressed in myocardial cells were used in this study. Zebrafish were maintained as described in the Zebrafish Handbook ( 28 ). All animal experiments were approved by the Animal Research Ethics Committee of Shanghai University of Traditional Chinese Medicine.…”

The PI3K/AKT Pathway—The Potential Key Mechanisms of Traditional Chinese Medicine for Stroke

“…Tanshinone IIA treatment promotes the recovery of brain function and increases neuronal viability; furthermore, the glucose concentration in the serum and cultured medium was increased, possibly regulated by an increased glucose uptake ability and activation of the PI3K/AKT/mTOR pathway ( 19 ). Sodium tanshinone IIA sulfonate is a water-soluble derivative of tanshinone IIA, which suppresses the expression of HIF-1 and hemoglobin genes via inhibition of the PI3K/AKT/mTOR pathway, thereby protecting against hypoxia in cerebral hemorrhage in zebrafish ( 20 ). Catalpol , an iridoid glycoside compound, the main active component of Rehmannia glutinosa Libosch (Di-Huang in Chinese), is commonly used in TCM for treating diseases of aging and stroke ( 21 ).…”

“…The neuroprotection of herbal single components through the PI3K/AKT pathway is detailed in the main text, some herbs contain multiple active components that synergistically protect the nervous system through PI3K/AKT-mediated downstream molecules. There are four active components of Salvia miltiorrhiza, including Sodium tanshinone IIA sulfonate, Sal A, SMND-309, and Cryptotanshinone, exert neuroprotection synergistically through the four PI3K/AKT-mediated pathways, including increasing neuronal viability, preventing apoptosis and promoting neurogenesis ( 20 , 56 , 59 ). Ginkgo biloba has various therapeutic effects, and contains multiple active components, like Ginkgolides, bilobalide, Ginkgetin aglycone.…”