To investigate the effect of Astragaloside Ⅳ on high-glucose-induced cardiomyocytes pyroptosis / 中华老年医学杂志

Objective:To investigate the effect of Astragaloside Ⅳ on high glucose-induced cardiomyocyte pyroptosis.Methods:H9c2 cells were cultured in vitro and divided into control group(5.5 mmol/L glucose), high glucose group(33.3 mmol/L glucose), Astragaloside Ⅳ group(33.3 mmol/L glucose+ 100μmol/L Astragaloside Ⅳ), and NLRP3 inhibitor group(33.3 mmol/L glucose+ 1μmol/L MCC950). Cell counting kit 8(CCK-8)was used to detect the activity of H9c2 cells.Lactate dehydrogenase(LDH)kit was used to detect the content of LDH in cell supernatant.Superoxide anion fluorescent probe(DHE)was used to detect the level of intracellular reactive oxygen species(ROS). Real-time fluorescence quantitative polymerase chain reaction(RT-qPCR)and Western blot were used to detect the mRNA and protein expression levels of pyroptosis-related genes.Immunofluorescence was used to detect the fluorescence intensity of NLRP3.Enzyme-linked immunosorbent assay(ELISA)was used to detect the level of inflammatory factors in cell supernatant.Results:When the concentration of Astragaloside Ⅳ was 100 μmol/L, it could significantly inhibit the decrease of cardiomyocyte viability induced by high glucose( P<0.01)and reduce LDH release( P<0.01). Compared with the control group, the level of ROS was increased( P<0.01), the mRNA and protein expressions of pyroptosis-related molecules were up-regulated( P<0.01 for all), the fluorescence intensity of NLRP3 was increased( P<0.01), and the levels of inflammatory factors in the cell supernatant were increased in the high glucose group( P<0.01). Compared with the high glucose group, the ROS level was decreased( P<0.01), the mRNA and protein expressions of pyroptosis-related molecules were down-regulated( P<0.05 or P<0.01), the fluorescence intensity of NLRP3 was decreased( P<0.01), and the levels of inflammatory factors in cell supernatant were decreased( P<0.05 or P<0.01)in Astragaloside Ⅳ group and inhibitor group. Conclusions:Astragaloside Ⅳ plays a protective role in high glucose-induced cardiomyocyte injury by inhibiting NLRP3/Caspase-1 signaling pathway and inhibiting pyroptosis.Moreover, it can improve the anti-inflammatory and antioxidant properties in cell models.

Study on steaming time and characteristic quality standard of salt-steaming Morinda officinalis based on Q-marker and anti-oxidative activities / 中国药房

OBJECTIVE To comprehensively screen the optimal steaming time of salt-steaming Morinda officinalis （SSMO） based on Q-markers and anti-oxidative activities， and to establish characteristic quality standard of the decoction pieces. METHODS The contents of six Q-markers （1-kestose， nystose， 1F-fructofuranosylnystose， inulotriose， inulotetraose and inulopentaose） in SSMO at different steaming time were determined by HPLC-ELSD method simultaneously. The activity of sample extracts to scavenge 4 kinds of oxidative free radical and their iron reduction abilities were determined by visible UV spectrophotometer. The optimal steaming time of SSMO was screened by gray relevance degree and entropy weight technique for order preference by similarity to an ideal solution （TOPSIS）-fusion model method. The contents of six Q-markers in 10 batches of SSMO prepared at the optimal steaming time were determined. HPLC-ELSD fingerprints of SSMO decoction pieces were established. RESULTS The results showed that the contents of six Q-markers were the highest when SSMO was steamed for 3-5 h； and the ability of scavenging DPPH·， ABTS·， PTIO·， ·OH and iron reduction ability was the best at 5 h. There were 20 common peaks in the fingerprints for 10 batches of samples， and the similarities were higher than 0.990. A total of 9 chromatographic peaks were identified， which were D-fructose （peak 1）， D（+）-glucose （peak 2）， sucrose （peak 3）， 1-kestose （peak 4）， nystose （peak 5）， 1F-fructofuranosylnystose （peak 6）， inulotriose （peak X2）， inulotetraose （peak X3） and inulopentaose （peak X4）. Average contents of six Q-markers were 4.17%， 5.54%， 6.60%， 2.89%， 2.62% and 2.13%， respectively. CONCLUSIONS The optimal steaming time of SSMO is 5 h； the contents of six Q-markers are primarily determined on the basis of dry product， which are no less than 3.03%， 4.11%， 4.87%， 2.15%， 1.96% and 1.58%， respectively. The ratio of Inulin-/Inulo oligosaccharides content is no more than 2.5.