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Objective@#To investigate the effects of hesperetin on fine particulate matter (PM2.5) induced apoptosis in H9c2 cells and related mechanisms.@*Methods@#H9c2 cells were divided into 4 groups: control group (cells were cultured without intervention), PM2.5 group (cells were treated with 800 µg/ml PM2.5), hesperetin group (H group, cells were treated by 40 µmol/L hesperetin for 1 h at 37 ℃), and hesperetin+PM2.5 group (H+PM2.5 group, cells were pretreated with hesperetin before PM2.5 treatment). Cells were cultured for corresponding interval. Apoptotic cells were detected by Annexin Ⅴ-FITC/PI apoptosis detection kit and Hoechst staining. The intracellular reactive oxygen species (ROS) levels were measured by DCFH-DA Fluorescence Probe and mitochondrial membrane potential (MMP) was detected with JC-1 staining, respectively in these groups. Apoptotic related protein and phosphorylated MAPK expression levels were determined by Western blot.@*Results@#(1) Flow cytometry results showed that the apoptosis rate of PM2.5 group ((48.94±3.20)%) was significantly higher than that of control group ((8.13±1.40)%, P<0.01), which was significantly reduced in H+PM2.5 group ((34.80±2.21)%) (P=0.003 2 vs. PM2.5 group, P<0.01 vs. control group). The number of Hoechst 33258 positive apoptotic cells was distinctly less in H+PM2.5 group than in PM2.5 group. (2) The ROS levels was significantly higher in PM2.5 group ((49.69±5.05)%) than in control group (10.57±1.33)%, P<0.01), which was significantly reduced in H+PM2.5 group ((35.08±3.90)%) (P=0.000 2 vs. PM2.5 group, P<0.01 vs. control group). (3) Green fluorescence indicating the JC-1 monomer form, which represented MMP loss of H9c2 cells, was significantly higher in PM2.5 group ((20.28±4.69)%) than in control group ((10.50±2.72)%, P<0.01), which was significantly decreased in H+PM2.5 group ((13.41±2.89)%) (P<0.01 vs. PM2.5 group, P=0.029 4 vs. control group). (4) The expression levels of Bcl-2 protein of H9c2 cells was lower in PM2.5 group ((76.94±4.52)%) than in control group (100%, P=0.000 9), which was significantly upregulated in H+PM2.5 group ((92.95±6.82)%) (P=0.027 5 vs. PM2.5 group, P=0.15 vs. control group). The expression levels of cleaved caspase-3 protein of H9c2 cells was significantly higher in PM2.5 group ((243.98±17.94)%) than in control group (100%, P=0.000 2), which was significantly downregulated in H+PM2.5 group ((200.45±4.31)%) (P=0.015 vs. PM2.5 group, P<0.01 vs. control group). (5) The expression levels of phosphorylated p38 MAPK protein of H9c2 cells was higher in PM2.5 group ((118.90±4.78)%) than in control group(100%, P=0.002 7), which could be significantly downregulated in H+PM2.5 group ((103.30±1.27)%) (P=0.01 vs. PM2.5 group, P=0.05 vs. control group). The expression levels of phosphorylated ERK protein of H9c2 cells was higher in PM2.5 group ((163.50±4.98)%) than in control group (100%, P<0.01), which was significantly downregulated in H+PM2.5 group ((139.10±2.72)%) (P=0.001 6 vs. PM2.5 group, P<0.01 vs. control group).@*Conclusions@#Hesperetin protects H9c2 cells from PM2.5 stimulation through reducing oxidative stress and protecting mitochondrial function, regulating the expression of apoptotic associated proteins as well as MAPK signal pathway, thus inhibiting H9c2 cells apoptosis.
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Objectives To investigate the relationship of sex difference with serum bisphenol-A (BP-A),adiponectin and metabolic syndrome (MetS)in elderly patients with isolated systolic hypertension(EISH).Methods A retrospective study of the clinical data was conducted in 540 subjects from the Cardiology and Geriatric Department in the Affiliated Hospital of Shanxi Medical University,Changzhi Municipal People,s Hospital and the Department of Cardiology of Shanxi Medical University,First Clinic Hospital from January 2010 to December 2013.Elderly patients with EISH were divided into male group(n=270)and female group(n=270).Meanwhile 560 older health persons were severed as controls,including 300 females and 260 males.The changes of BP-A and adiponectin (Ad) concentration were measured.The blood lipid,insulin resistance index (HOMA-IR),blood pressure,body mass index,heart rate variability and ultrasonic change of heart and blood vessel were tested regularly.Results The level of serum BP-A[(0.89±0.10)ng/L vs.(0.57±0.04)ng/L]and [(0.64±0.10)ng/L vs.(0.55 ± 0.08)ng/L] were higher in male EISH vs in male control,and in female EISH than in female control (F =23.76,23.86,all P ＜ 0.01),respectively.The levels of adiponectin were lower in male EISH vs control[(4.9±1.4)ng/L vs.(10.5±2.7)ng/L and in female EISH vs control(6.0±1.3) ng/L vs.(11.5±3.3)ng/L),F=13.10,16.20,all P＜0.01.Root mean sequare of the successive normal sinus RR interval difference(rMSSD)were lower in male/ female EISH than control groups(F=13.10、13.70,P ＜0.01).Serum BP-A level was positively correlated with the bocly mass index and systolic pressure (r =0.38,0.54,P ＜ 0.01),and was negatively correlated with serum Ad and rMSSD(r=-0.46,-0.42,P＜0.01).Conclusions Obvious gender difference in changes of serum BP-A exists in older patients with EISH.Network cytokines may take part in the pathophysiological process of the obesity related hypertension.
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Objective@#To investigate the effect of natural active compounds apigenin (API) on the proliferation of rat aortic vascular smooth muscle cells (VSMCs) induced by lipopolysaccharide (LPS) and related mechanisms.@*Methods@#VSMCs of primary cultured SD rats were obtained and the cytotoxic effects of API (0, 10, 20, 40 and 80 μmol/L) was explored by CCK-8 method. Impact of LPS (0, 0.1, 1, 10 and 100 μg/ml) on VSMCs proliferation and the impact of API (0, 10, 20, 40 μmol/L) on LPS (10 μmol)-induced VSMCs proliferation by CCK-8 methods. Using EdU and FCM method, we observed the effect of API on proliferation of VSMCs induced by LPS. VSMCs proliferation and cell cycle were also assessed by EdU method and FACS in 10 μg/ml LPS, 10 μg/ml LPS+ 40 μmol/L API and equal volume DMSO treated VSMCs.@*Results@#(1) CCK-8 cell vitality test showed that cell vitality was not affected by 0-40 μmol/L API, while cell vitality was significantly reduced by 80 μmol/L API (57%), which was significantly lower than in blank group (P<0.05). (2) VSMCs proliferation was significantly promoted by 0.1, 1 and 10 μg/ml LPS and peaked in 10 μg/ml LPS stimulated VSMCs group, while VSMCs proliferation was significantly reduced in 100 μg/ml LPS stimulated group (P<0.05 vs. blank group). (3) LPS (10 μm/ml) induced VSMCs proliferation was not affected by 10 μmol/L API, which was significantly inhibited by 20 and 40 μmol/L API (both P<0.05 vs. LPS). (4) VSMCs proliferation assessed by EdU was significantly higher in LPS group than in blank group (P<0.01), which could be significantly reduced by cotreatment with API (P<0.01). (5) FACS results showed that percent of VSMCs in G0/G1 stage was significantly lower in LPS group compared to blank group (P<0.05), which could be significantly increased post API treatment (P<0.05 vs. LPS), while percent of VSMCs in S stage was significantly lower post API treatment in comparison with LPS group.@*Conclusion@#API can significantly inhibit LPS-induced proliferation of VSMCs, partly through inhibiting mitosis and inducing G0/G1 cell cycle arrest.
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Objective To investigate the protective effects of antioxidant taurine plus metoprolol on the blood pressure(BP)and blood vessel function of elderly spontaneously hypertensive rats(SHR).Methods SHRs were divided into three groups (6 rats,each):metoprolol group (100 mg · kg-1 · d-1,intragastric administration);taurine group (200 mg · kg-1 · d-1,intraperitoneal injection);taurine plus metoprolol group(metoprolol 100 mg· kg-1 · d-1,intragastric administration ± taurine 200 mg· kg-1 · d-1,intraperitoneal injection).The control group[6 Wistar-Kyoto(WKY) rats]was treated with the same volume of sterile normal saline on the same schedule in intragastric administration and intraperitoneal injection.Blood pressure variability(BPV),diurnal variation of BP,the level of glutathion peroxidase (GSH-Px),malonaldehyde (MDA) and the aorta GSTM1 enzyme expression were evaluated before and 14 days after treatment.Results The serum GSH-Px activities were higher in metoprolol group[(2 759.8 ± 117.6) kU/L],taurine group [(2 848.0 ± 280.2) kU/L] and taurine plus metoprolol group[(3 052.8±283.7)kU/L]than in control group[(2 368.0± 60.4) kU/L] (all P＜0.05).Fourteen days after treatment,MDA level was significantly lower in taurine group[(9.5±0.7)ng/L,P＜0.01]than in control group[(13.7±1.5)ng/L].However,there was no statistical difference in MDA level between metoprolol group/taurine plus metoprolol group and control group.Forty weeks after treatment with taurine,GSTM1 protein expression was increased,but it had no statistical difference.Conclusions The combined antioxidant taurine and metoprolol can effectively decrease BPV of elderly SHRs,in which the antioxidant effect might be associated with elevated GSTM1 protein expression.