Abstract
Trichosanthis Pericarpium (TP) is a traditional Chinese medicine for treating cardiovascular diseases. In this study, we investigated the effects of TP aqueous extract (TPAE) on hypoxia/reoxygenation (H/R) induced injury in H9c2 cardiomyocytes and explored the underlying mechanisms. H9c2 cells were cultured under the hypoxia condition induced by sodium hydrosulfite for 30 min and reoxygenated for 4 h. Cell viability was measured by MTT assay. The amounts of LDH, NO, eNOS, and iNOS were tested by ELISA kits. Apoptotic rate was detected by Annexin V-FITC/PI staining. QRT-PCR was performed to analyze the relative mRNA expression of Akt, Bcl-2, Bax, eNOS, and iNOS. Western blotting was used to detect the expression of key members in the PI3K/Akt pathway. Results showed that the pretreatment of TPAE remarkably enhanced cell viability and decreased apoptosis induced by H/R. Moreover, TPAE decreased the release of LDH and expression of iNOS. In addition, TPAE increased NO production and Bcl-2/Bax ratio. Furthermore, the mRNA and protein expression of p-Akt and eNOS were activated by TPAE pretreatment. On the contrary, a specific inhibitor of PI3K, LY294002 not only inhibited TPAE-induced p-Akt/eNOS upregulation but alleviated its anti-apoptotic effects. In conclusion, results indicated that TPAE protected against H/R injury in cardiomyocytes, which consequently activated the PI3K/Akt/NO signaling pathway.
Highlights
Ischemic heart disease, which includes coronary heart disease (CHD), is one of the leading causes for death all over the world [1,2,3,4]
The results revealed that cell viabilities did not decrease among TP aqueous extract (TPAE)-treated groups compared with the normal group
Our results indicate that recoupling eNOS and reducing inducible NOS (iNOS) collectively regulate the amount of Nitric oxide (NO) during the hypoxia/ reoxygenation (H/R)
Summary
Ischemic heart disease, which includes coronary heart disease (CHD), is one of the leading causes for death all over the world [1,2,3,4]. Angina pectoris is the fatal subtype of CHD, resulting from myocardial ischemia and hypoxia [5]. The effective therapy for relieving ischemia is early myocardial reperfusion or primary percutaneous coronary intervention (PCI) [6,7]. Both of them may self-contradictorily lead to further injury, which is known as myocardial ischemia/reperfusion injury (MI/RI) [7,8]. It has been reported that TP protects from ischemia reperfusion injury, hypoxia, angina and various cardiovascular diseases in China [4,5,14]. Many studies have verified that TP displayed its protective effect on MI/RI [5,13,14,15]
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