1.Side chain carbon losssalidroside analogues (N04) protects EAhy926 human endothelial cells against hypoxia injury by regulating the expressions of HIF-1α-related factors
Jing BU ; Yongliang ZHANG ; Lingzhi LI ; Zhiheng WU ; Pei DONG ; Jianyu LI ; Yaxiao LI
Journal of Medical Postgraduates 2017;30(8):798-803
Objective Salidroside is a major active component of integripetal rhodiola herbal medicine, which has a significant activity against hypoxia and ischemia.This study was to investigate the effects of side chain carbon losssalidroside analogues (N04) on the expressions of HIF-1α-related factors in the hypoxia-injured EAhy926 human vascular endothelial cells.Methods EAhy926 human umbilical vein endothelial cells in the logarithmic growth phase were randomly divided into a normal control, a hypoxia model control, a salidroside, a high-dose N04, a medium-dose N04, and a low-dose N04 group.The hypoxia model was established by depriving the culture medium of sugar and serum and culturing the EAhy926 cells in an environment of 95%N2+5%CO2 for 2 hours, followed by intervention with salidroside at 1×10-6 mol/L and N04 at 1×10-6, 1×10-7, and 1×10-8 mol/L, respectively.Then, the activity of the cells was detected by MTT assay, their LDH activity examined by spectrophotometry, the mRNA expressions of HIF-1α and VEGF measured by RT-PCR, the protein expressions of HIF-1α, VEGF and pVHL determined by Western blot, and the activity of eNOS measured by ELISA.Results Compared with the normal control group, the hypoxia model cells showed significantly reduced activity (0.51±0.05 vs 0.27±0.02, P<0.01), an elevated LDH level ([6.65±1.43] vs [78.82±2.33] U/L, P<0.01), and decreased eNOS activity ([1.56±0.23] vs [1.16±0.20] U/100 mL, P<0.01).In comparison with the hypoxia model group, the cells treated with high-, medium-, and low-dose N04 exhibited remarkably increased activity (0.27±0.02 vs 0.0.42±0.05, 0.40±0.03 and 0.37±0.04, P<0.01), a reduced LDH level ([78.82±2.33] vs [53.05±3.90], [58.42±4.45] and [62.73±3.63] U/L, P<0.01), and increased eNOS activity ([1.16±0.20] vs [3.01±0.47], [2.60±0.26] and [2.32±0.29] U/100 mL, P<0.01).The activity of eNOS was also increased in the salidroside group ([2.32±0.29] U/100 mL, P<0.01).The cell activity in the high-and medium-dose N04 groups was markedly higher than that in the salidroside group (P<0.05), and so was the eNOS activity in the high-dose N04 group and the LDH level in the medium-and low-dose N04 groups (P<0.05).In comparison with the normal control group, the expressions of HIF-1α mRNA, HIF-1α protein and VEGF protein were significantly up-regulated in the hypoxia model group (P<0.01) while that of the pVHL protein markedly down-regulated (P<0.01).Compared with the hypoxia model group, the expressions of HIF-1α mRNA, HIF-1α protein and VEGF protein were remarkably reduced (P<0.05), while that of the pVHL protein markedly elevated (P<0.05).Both the expressions of VEGF mRNA and HIF-1α protein were significantly lower in the medium-and low-dose N04 groups than in the salidroside group (P<0.05).Conclusion N04 can protect vascular endothelial cells against hypoxia-induced injury by regulating the expression of HIF-1α-related factors and eNOS.
2.Plasma metastin in adolescent polycystic ovary syndrome
Xiaoli CHEN ; Yaqin MO ; Lin LI ; Yaxiao CHEN ; Yu LI ; Qingxue ZHANG ; Dongzi YANG
Chinese Journal of Obstetrics and Gynecology 2009;44(10):745-749
Objective This study was designed to investigate the correlationship between plasma metastin and pathogenesis of adolescent women with polycystic ovary syndrome(PCOS).MethodsFrom Jan 2006 to Jun.2006.42 PCOS patients including 19 adolescent women and 23 adults with syndrome were treated in Second Affiliated Hospital of Sun Yat-Sen University.According to the range of age,those patients were divided into 19 cases in adolascent group(≤19 years)and 23 cases in aduh group(>19 years).Meanwhile,20 adolescent women were matched as controls.Blood samples were collected between day 1 and day 5 of a spontaneous bleeding episode in the PCOS groups and a menstrual cycle of the controls.The Jevels of luteinizing hormone(LH),follicle-stimulating hormone(FSH),testosterone(T),free T(FT),dehydroepiandrosterone sulfate(DHEAS),sex hormone-binding globulin(SHBG),insulin,glucose,and metastin were detected from day 1 to day 5 of spontaneous bleeding or withdrawal bleeding by progesterone.On the next day,oral glucose tolerance test(75 g)and insulin release test were performed on those above patients and controls.The area under carve(AUC),the ratio of fasting blood glucose to insulin(GIR)and homeostasis model assessment insulin resistance jndex ( HOMA-IR)were calculated.Results(1)The level of hormone:the level of LH was in(12±7)U/L in adult group and(12±8)U/L in adolescent PCOS group,which were significantly higher than(6±4)U/L in controls(P<0.05).The level of FT was(2.3±1.2)pmol/L in adult group,which was significantly higher than(1.3±0.8)pmol/L in adolescent group and(1.1±0.5)pmol/L in control roup(P<0.05).It was observed that the level of(3.1±2.7)μmol/L in adolescent group was significantly lower than(6.3±2.7)μmol/L in control group(P<0.05).Meanwhile,the level of FAI of 5.6±4.1 in adult group was significantly higher than 3.0±1.3 in control group(P<0.05).No significant difference in FSH,T and SHBG levels among three groups were observed (P>0.05).(2)Metastin and metabolism:Both the levels of fasting blood insulin,2-hour insulin and AUC of insulin were(13±7)mU/L,(88±59)mU/L and(133±80)mU·L-1·min-1 in adolescent group,which were significantly higher than(7±3)mU/L,(57±29)mU/L and(82±34)mU·L-1·min-1 in control group.The fasting blood insulin of(13±7)mU/L in adolescent group was significantly higher than (9±5)mU/L in adult group.The level of fasting blood glucose and 2-hour glucose were(5.01±0.44)mmol/L and(6.48±1.16)mmol/L in adult group,which were significantly higher than(4.68±0.29)mmol/L and(5.44±0.83)mmol/L in control group and(4.67±0.30)mmol/L and(5.93±1.44)mmol/L in adolescent group.The glucose AUC of(9.99±1.85)mmol·L-1·min-1 in adult group was significantly higher than(8.42±1.53)mmol·L-1·min-1 in control group(P<0.05).HOMA-IR of 2.6±2.0 in adolescent group was significantly higher than 1.4±0.7 in control group.GIR of 10±8 in adolescent group was significantly lower than 16±10 in control group(P<0.05).The metastin level of (0.25±0.19)pmol/L in adolescent group and(0.29±0.29)pmol/L in adult group were all significantly higher than(0.18±0.23)pmol/L in control group(PPh glucose were observed(r=0.256,0.286 and 0.267.P=0.044.0.025 and 0.043).Conclusions The expression of metastin in adolescent PCOS women was significantly higher that of normal adolescent women The increased level of metastin might be associated with pathogenesis of adolescent women with PCOS.
3.Bone marrow mesenchymal stem cell transplantation for treatment of emphysema:intravenous versus intratracheal approach
Xiaojian ZHAO ; Caiping LU ; Weiwei CHU ; Yaxiao ZHANG ; Bing ZHANG ; Qiang ZHEN ; Guoliang TAN ; Renfeng WANG ; Jiabao LIU ; Lin WU
Chinese Journal of Tissue Engineering Research 2015;(14):2211-2215
BACKGROUND:Bone marrow mesenchymal stem cel s transplantation can inhibit experimental emphysema inflammatory reaction and apoptosis, and has been experimental y confirmed to treat severe lung function impairment. OBJECTIVE:To explore the inhibitory effects of bone marrow mesenchymal stem cel s transplantation via different ways on inflammatory reaction and apoptosis due to experimental emphysema. METHODS:Female Wistar rats were randomly divided into control group, intravenous group and endotracheal group fol owing model establishment using fumigation plus intratracheal instil ation of porcine pancreatic elastase. In the latter two groups, bone marrow mesenchymal stem cel s from male rats were injected via the tail vein and the trachea, respectively. In the control group, rats were given PBS via he tail vein and trachea. At 14 days after transplantation, pathological changes of rat lung tissues were observed, cel apoptotic index in alveolar wal cel s and tumor necrosis factorαlevel in the bronchoalveolar lavage fluid were detected. RESULTS AND CONCLUSION:Compared with the control group, in the intravenous and endotracheal groups,the pathological changes of lung tissues were relieved, tumor necrosis factorαlevel and apoptosis index were reduced significantly (P<0.01);but there were no differences between the intravenous and endotracheal groups (P>0.05). These findings indicate that bone marrow mesenchymal stem cel s transplantation via the tail vein and trachea both can exert obvious therapeutic effects on emphysema. Moreover, cel transplantation via the tail vein is more convenient and easier than that via the trachea in the treatment of emphysema.
4.Bone marrow mesenchymal stem cell transplantation suppresses emphysema-induced inflammation and apoptosis
Xiaojian ZHAO ; Caiping LU ; Weiwei CHU ; Qiang ZHEN ; Guoliang TAN ; Yaxiao ZHANG ; Renfeng WANG ; Bing ZHANG ; Jiabao LIU
Chinese Journal of Tissue Engineering Research 2014;(6):906-911
BACKGROUND:Bone marrow mesenchymal stem cells transplantation can change the surrounding microenvironment through paracrine mechanisms, and can be employed for treatment of serious damage to lung function through the promotion of angiogenesis, inhibition of apoptosis and maintaining functional stability of autonomic nervous system. OBJECTIVE:To observe the inflammatory reaction in experimental emphysema and inhibition of apoptosis through bone marrow mesenchymal stem cells transplantation.
METHODS:Twenty-four Wistar female rats were randomly divided into three groups:healthy control group, model group and experimental group. In the latter two groups, smoking and endotracheal instil ation of porcine pancreatic elastase were performed to establish emphysema models. After modeling, bone marrow mesenchymal stem cells were injected via tail vein in the experimental group. Pathological changes of the lung, the level of tumor necrosis factor-alpha and cellnumber in the bronchoalveolar lavage fluid as wel as apoptotic index in lveolar wal s were detected after celltransplantation.
RESULTS AND CONCLUSION:In the model and experimental groups, pathological changes of lung tissues were observed to different extent. The lung pathological changes were slighter in the experimental group than the model group (P<0.01). The level of tumor necrosis factor-alpha and apoptotic index in lung tissue were lower in the experimental group than the model group (P<0.01). These findings indicate that bone marrow mesenchymal stem cells can improve emphysema pathological y through inhibition of inflammatory response and apoptosis in experimental emphysema.