Objective To observe the therapeutic effect of Di'ao Xinxuekang (DAXXK) on myocardial ischemia-reperfusion injury in rats, and to explore its mechanisms. Methods The myocardial ischemia-reperfusion injury model was established by the ligation of descending coronary artery in rats. Then animals after the modeling were randomized into model group, DAXXK-d (31.5 mg/kg) group, DAXXK-g (63.0 mg/kg) group and Diltiazem (24.8 mg/kg) group. A separate sham group was used as control. The treatment group was given DAXXK once a day for 7 days. Cardiac function and cardiac configuration were measured by color Doppler ultrasound diagnostic method. Hemodynamics was measured by Millar catheter method. The arterial oxygen saturation and blood oxygen pressure were measured by i-STAT 300 blood gas analyzer. Inflammatory cytokines interleukin (IL)-1β, IL-6, tumor necrosis factor (TNF)-α, adhesion molecules VCAM-1, ICAM-1, and apoptosis-related protein Bcl-2, Bax were detected by ELISA. Myocardial apoptosis was measured using TUNEL method. Results Compared with model group, the left ventricular fractional shortening (FS), the systolic and diastolic function were improved, and the left ventricular pressure maximum rise/ fall rates (± LVdp/dtmax) were increased, in DAXXK group. DAXXK improved lung function, increased arterial oxygen pressure and oxygen content. The inflammatory cytokines IL-1β, IL-6, TNF-α and adhesion molecules ICAM-1 and VCAM-1 were decreased in DAXXK group. The myocardial swelling and inflammatory infiltration were relieved, myocardial apoptosis was reduced, the expression of Bcl-2 protein was increased and the expression of Bax protein was decreased in DAXXK group. Conclusion DAXXK can protect myocardial ischemia-reperfusion injury, which involved in the inhibition of apoptosis and reduction of inflammatory cytokines.

Objective To evaluate the expression of Tc17 in a cigarette smoke-induced mice model of emphysema.To explore the probable mechanisms about how Tc17 cells to elevate in lungs of mice.Methods Forty male Balb/c mice were randomly divided into four groups,including control group ( 12 weeks,C12),control group (24 weeks,C24),smoke-exposure group (12 weeks,S12) and smoke-exposure group (24 weeks,S24 ),10 mice each group,Emphysema of mice was observed by HE pigmentation.Morphological changes were evaluated by mean linear intercepts (Lm) and destructive index (DI).The proportion of CD8+ IL-17 + Tc17,CD8+ IL-17 + CC chemokine receptor type 6 ( CCR6 ) + and 6CCR6 + Tc17 cells in lungs of mice was determined by flow cytometry.The mRNA expressions of retinoidrelated orphan nuclear receptor(RORγt) and IL-17 were evaluated by real-time PCR.The levels of IL-1 β,IL-6,IL-23,transforming growth factor β (TGFβ) and CC chemokine ligand 20 (CCL20) were tested by ELISA.Correlations among these indexes were analyzed.Results Lm and DI were significantly higher in S12 and S24 than in C12 and C24,S24 in particular (t value 4.378-15.188,all P ＜ 0.05).The percentages of Tc17 in S12 and S24[(9.28 ± 1.12)%,( 13.13 ±3.56)%]was significantly increased as compared with that in C12 and C24[(2.40 ±0.60 )%,(2.64 ±0.96 )%],S24 in particular.The mRNA levels of RORγt and IL-17 in S12 and S24 were higher than in C12 and C24,S12 and S24 in particular.There was significant difference (all P ＜0.05 ).The frequency of Tc17 cells had a positive correlation with Lm and DI ( r value were 0.734 and 0.884 respectively,P ＜ 0.01 ).The percentages of CD8+ IL-17 + CCR6 +T cells and CCR6 + Tc17 were significantly elevated in S12 and S24 compared to C12 and C24,S24 in particular (all P ＜ 0.05 ).There was positive correlation between Tc17 cell ratio and CCL20 levels( r =0.899,P ＜0.01 ).The levels of IL-1 β,IL-6,IL-23 and TGFβ in S12 and S24 were significantly increased as compared with that in C12 and C24.There was significant difference (all P ＜0.05).Meanwhile,the frequency of Tc17 cells had a positive correlation with IL-1β,IL-6,IL-23,and TGFβ.Conclusions An up-regulation of proportions Tc17 in lungs of cigarette smoke-induced emphysema mice were detected.The CCR6/CCL20 axis and the increased IL-1β,IL-6,IL-23 and TGFβ probably contributed to this up-regulation.

Objective To investigate the effect of cigarette smoke exposure on the interleukin-17-producing CD4+T helper lymphocyte( Th17 ) in peripheral blood and lungs of cigarette smoke-induced emphysema mice. MethodsForty male BALB/c mice were randomly divided into 4 groups, including a control group C12, a control group C24, a smoke-exposure group S12 and a smoke-exposure group S24, 10 mice each group. Morphological changes were evaluated by mean linear intercepts (Lm) and destructive index (DI). The concentration of IL-17, IFN-γ and TNF-α in bronchoalveolar lavage fluid(BALF) and the concentration of IFN-γ and TNF-α in lung homogenates were measured by enzyme-linked immunosorbent assay (ELISA). The proportion of CD4+IL-17+ Th17 cells in peripheral blood and lungs of mice was determined by flow cytometry. The mRNA expressions of RORγt and IL-17 in peripheral blood and lungs were evaluated by real-time PCR. ResultsLm and DI were significantly higher in S12 and S24 groups [(39.19±3.51)μm vs (46.87±7.16) μm, 39.13±1.57 vs 45.16±3.13]than that in C12 and C24 groups [(32.60± 3.21 ) μm vs (32.38±3.73) μm, 28.23± 1.62 vs 28.86±2.07], all P＜0.05. IL-17, IFN-γand TNF-α in BALF of S12 and S24 groups [(119.72±10.72) ng/L vs (296.40±14.00) ng/L,(129.7±22.2) ng/L vs(251. 1±62.4) ng/L, (17.35±1.60) ng/L, (36.35±1.43) ng/L]were higher than those in C12 and C24 groups [(52.06±4.70) ng/L vs (51.89 ±6.82) ng/L, ( 85.8 ±26.8) ng/L vs ( 88.9 ± 11.5 ) ng/L,(6.41 ±0.90) ng/L vs (5.85±0.92) ng/L], IFN-γ and TNF-α in lungs of S12 and S24 groups [( 1124.3±174.4) ng/L vs (1342.7±206.1) ng/L, (77.2±13.7) ng/L vs (101.7±19.0) ng/L, (129.7±22.2)ng/L vs (251.1 ±62.4) ng/L]were higher than those in C12 and C24 groups[(946.2±81.9) ng/L vs (1027.2±188.3) ng/L, (62.1±16.1) ng/L vs(64.4±15. 1) ng/L], all P＜0.05. The percentage of Th17 cells in lungs of S12 and S24 groups [(3.27±1.12)％ vs (7.19±2.24)％, ( 1.96±0.61 )％ vs (3.82±1.26)％]was significantly increased as compared with that of C12 and C24 groups [(1.80± 0.75)％ vs (1.99±0.59)％], all P＜0.05. And the percentage of Th17 cells in peripheral blood of S12 and S24 groups [(1.96±0.61)％ vs ( 3.82± 1.26)％]was also significantly higher than those in C12 and C24 groups [(0.90±0.37)％ vs (0.97±0.32)％], all P＜0.05. In peripheral blood and lungs,the mRNA levels of RORγt and IL-17 in S12 and S24 groups were higher than in C12 and C24 groups, all P＜0.05.Moreover, the frequency of Th17 cells in peripheral blood and lungs had a positive correlation with Lm and DI( r=0.706-0.772 ,all P＜0.05 ). ConclusionAn up-regulation of proportions Th17 in peripheral blood and lungs of cigarette smoke-inducsed emphysema mice were detected. It may be involved in the amplified and persistent inflammation.