Mechanism of Trichosanthis Pericarpium Aqueous Extract in Protecting H9c2 Cardiomyocytes from Hypoxia/Reoxygenation Injury via PI3K/Akt/NO Signaling Pathway
10.13422/j.cnki.syfjx.20192104
- VernacularTitle: 瓜蒌皮提取物基于PI3K/Akt/NO信号通路保护缺氧/复氧损伤心肌细胞的机制
- Author:
Dong-hai CHU
1
;
Zhen-qiu ZHANG
2
Author Information
1. School of Biomedical and Chemical Engineering, Liaoning Institute of Science and Technology, Benxi 117004, China
2. School of Pharmacy, Liaoning University of Traditional Chinese Medicine, Dalian 116600, China
- Publication Type:Research Article
- Keywords:
Trichosanthis Pericarpium aqueous extract;
hypoxia/reoxygenation injury;
cardiomyocytes;
phosphatidylionsitol-3-kinase/protein kinase B/nitric oxide(PI3K/Akt/NO)
- From:
Chinese Journal of Experimental Traditional Medical Formulae
2019;25(22):42-48
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To investigate the effect of Trichosanthis Pericarpium aqueous extract(TPAE)in protecting H9c2 cells from hypoxia/reoxygenation (H/R) injury by activating phosphatidylionsitol-3-kinase/protein kinase B/nitric oxide(PI3K/Akt/NO) signaling pathway. Method: The 2.5 mmol·L-1 Na2S2O4 was used to induce the model of H9c2 cardiomyocytes H/R injury in the experiments. The cultured H9c2 cardiomyocytes were randomly divided into normal group, H/R group (model group) and inhibition group (LY294002, 10 μmol·L-1). In the H/R + TPAE group, 50 mg·L-1 TPAE was added to the cultures at 24 h before H/R exposure. Cell viability was measured by methyl thiazolyl tetrazolium (MTT) assay. The amounts of NO, endothelial nitric oxide synthase (eNOS), and induced nitric oxide synthase (iNOS) were tested by enzyme linked immunosorbent assay (ELISA) kits. Reverse transcription-quantitative real-time polymerase chain reaction (Real-time PCR) was performed to analyze relative mRNA expressions of Akt, eNOS and iNOS. Western blot was used to detect the expressions of Akt, p-Akt (Ser 473), eNOS, and iNOS. Result: Compared with the normal control group,the cell viability significantly decreased in the model control group (P<0.01), the release of NO was obviously down-regulated (P<0.01), the mRNA and protein expressions of p-Akt, Akt, eNOS were remarkably decreased (P<0.01), while those of iNOS were up-regulated (P<0.01). Compared with the H/R group,the pretreatment with TPAE remarkably improved the morphological lesion of cardiomyocytes, enhanced cell viability(P<0.01),increased the expressions of Akt, p-Akt (Ser 473) and eNOS(P<0.01),decreased the expression of iNOS(P<0.01), and increased the release of NO(P<0.01). The PI3K inhibitor LY294002 was used. Obviously, cardioprotection of Trichosanthis Pericarpium aqueous extract was blocked by co-treatment LY294002, and cell viability was correspondingly reversed. Conclusion: TPAE can protect H9c2 cardiomyocytes from H/R injury by activating PI3K/Akt signaling pathway,which might be related to the up-regulation of the mRNA and protein expressions of eNOS, the down-regulation of the level of iNOS, and the increase of the production of physiological amounts of NO.
