Myocardial infarction and following reperfusion therapy-induced myocardial ischemia/reperfusion (I/R) injury have been recognized as an important subject of cardiovascular disease with high mortality. As the antiarrhythmic agent, Wenxin Granule (WXG) is widely used to arrhythmia and heart failure. In our pilot study, we found the antioxidative potential of WXG in the treatment of myocardial I/R. This study is aimed at investigating whether WXG could treat cardiomyocyte hypoxia/reoxygenation (H/R) injury by inhibiting oxidative stress in mitochondria. The H9c2 cardiomyocyte cell line was subject to H/R stimuli to mimic I/R injury in vitro. WXG was added to the culture medium 24 h before H/R exposing as pretreatment. Protein kinase C-δ (PKC-δ) inhibitor rottlerin or PKC-δ lentivirus vectors were conducted on H9c2 cells to downregulate or overexpress PKC-δ protein. Then, the cell viability, oxidative stress levels, intracellular and mitochondrial ROS levels, mitochondrial function, and apoptosis index were analyzed. In addition, PKC-δ protein expression in each group was verified by western blot analysis. Compared with the control group, the PKC-δ protein level was significantly increased in the H/R group, which was remarkably improved by WXG or rottlerin. PKC-δ lentivirus vector-mediated PKC-δ overexpression was not reduced by WXG. WXG significantly improved H/R-induced cell injury, lower levels of SOD and GSH/GSSG ratio, higher levels of MDA, intracellular and mitochondrial ROS content, mitochondrial membrane potential and ATP loss, mitochondrial permeability transition pore opening, NOX2 activation, cytochrome C release, Bax/Bcl-2 ratio and cleaved caspase-3 increasing, and cell apoptosis. Similar findings were obtained from rottlerin treatment. However, the protective effects of WXG were abolished by PKC-δ overexpression, indicating that PKC-δ was a potential target of WXG treatment. Our findings demonstrated a novel mechanism by which WXG attenuated oxidative stress and mitochondrial dysfunction of H9c2 cells induced by H/R stimulation via inhibitory regulation of PKC-δ/NOX2/ROS signaling.

To study the correlations and relationship between spore yield and various macromorphological traits of Ganoderma lingzhi, a field experiment was conducted in a randomized block design with 3 replications. Ten macromorphological traits, including pileus diameter, pileus crust thickness, context thickness, tube thickness, pileus thickness, stipe length, stipe diameter, stipe weight, pileus weight, and spore yield, were recorded for all of the tested strains. There was significant variation among the strains for all of the traits studied. The results indicated that the highest variation was observed in spore yield and pileus weight. Correlation studies revealed that among 9 macromorphological traits, only the pileus weight of the fruiting body was significantly positively correlated with spore yield (r2 = 0.674*). Pileus diameter showed significant positive association with pileus weight of fruiting body (r2 = 0.838*). Stepwise multiple regression analysis showed that pileus weight and spore yield had a linear relationship (spore yield = -21.95 + 1.51 * pileus weight). The coefficient of determination of stepwise regression analysis (r2 = 0.4543) revealed 45.43% variation in the spore yield because of its relationship with pileus weight. Regression coefficient (b = 1.51) showed that a unit (1 g) increase in the pileus weight of the fruiting body resulted in a proportional increase of 1.51 g in spore yield. The derived information would be very useful when selecting potentially breeding strains for future G. lingzhi improvement programs.