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AIM:To explore whether YAP protein is important in induced pluripotent stem cell ( iPSC)-induced cardiovascular progenitor cell and/or vascular smooth muscle differentiation .METHODS:Using episomal vector based reprogramming , we generated human iPSCs from donor fibroblasts .We used both this iPSCs and human H 1 embryonic stem cells to differentiate into vascular smooth muscle cells (VSMCs) through cardiovascular progenitor cells (CVPC).Western blotting, qPCR and immunofluorescence microscopy were used to check the expression of YAP and related genes during this differentiation process .RESULTS:The results showed that iPSCs expressed pluripotent stem cell markers, such as Oct4, Nanog, Sox2, TRA-1-60 and SSEA3, and could form teratoma in SCID mice.YAP was highly expressed in pluripotent stem cells , but dramatically decreased when CVPC differentiation started .YAP gradually increased dur-ing CVPC three-day differentiation.The TAZ and YAP binding partner TEAD1, but not TEAD2 and TEAD4, have similar expression pattern in CVPC differentiation .Immunofluorescence result confirmed that YAP was activated and accumulated in nucleus .Interesting-ly, both YAP and phosphorylated YAP expression decreased to very low level after CVPC differentiated into VSMCs in 7 days.TEAD4 and TAZ also decreased, while TEAD1, TEAD2 and TEAD3 expression did not change during VSMC differentiation .CONCLU-SION:YAP and TEAD1 expression increased during CVPC differentiation , while YAP and TEAD4 expression decreased from CVPC to VSMCs differentiation , which suggested YAP might have different function during diverse cell differentiation .
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Objective To observe changes of spatial learning-memory in rats with chronic hypoxic hypercapnia and the effect of gingkgo biloba extra. Methods After established the rat model of chronic hypoxic hypercapnia,seventy-two rats were randomly divided into four groups normal control (NC),hypoxic-hypercapnia 4-week (4HH),hypoxic-hy-percapnia 4-week+gingkgo biloba extra (EGb)high dose(100 mg/kg)group[4HH+EGb(H)] and hypoxic-hypercapnia 4-week+EGb low dose (50 mg/kg) group[4HH+EGb(L)]. Praxiology in rats was asessed by the Morris water maze and step down test. Results The spatial learning-memory in rats exposed to chronic hypoxic-hypercapnia 4-week(4HH group)were displayed significant impairment in their performance,the longer mean escape latencies and swim path dis tances,the more error times. 4HH+EGb(H) and 4HH+EGb(L)groups shortened the reaction time of leaning, pro longed the latent time of memory, reduced times of mistakes. Conclusions EGb can enhance the capacity of learning-memory in the rats exposed chronic hypoxic hypercapnia.
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AIM: To explore the effects and possible mechanism of exogenous spermine on the apoptosis of primary cultured neonatal cardiomyocytes induced by simulated ischemia-reperfusion (I/R) injury. METHODS: To establish a model of simulated I/R, the primary cultured neonatal rat cardiomyocytes were incubated in ischemia-mimetic solution (under the conditions of hypoxia plus serum deprivation) for 2 h, and re-incubated the cells in normal culture medium for 24 h. The apoptotic cell death was assayed by flow cytometry. The morphological alterations of the cells were observed under transmission electron microscope. The transcription and expression of Fas and FasL were determined by the methods of RT-PCR, Western blotting and immunofluorescence. RESULTS: The cells exposed to I/R underwent significant apoptosis, and the percentage of apoptotic cells was 27.4%±1.8%, much higher than that in normal group (5.7%±0.3%). In I/R group the evident histopathological changes were observed and the myocardial transcription and expression of Fas and FasL were significantly upregulated. Compared to normal group, mRNA expression of Fas and FasL increased 2.2 folds and 2.4 folds, respectively, and their proteins increased 1.7 folds and 1.9 folds at 24 h of reperfusion respectively (P<0.01). Pretreatment with 10 μmol/L spermine significantly inhibited apoptosis of I/R injured cells, and the percentage of apoptotic cells was 21.7%±1.3% (P<0.01, as compared to I/P group). Spermine also suppressed the expression of Fas and FasL significantly (P<0.05 or P<0.01, as compared to I/P group). CONCLUSION: Spermine plays anti-apoptotic effect on the cultured neonatal myocardial cells under the condition of I/R injury by suppressing the expression of Fas/FasL.
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<p><b>OBJECTIVE</b>To examine the change of puerarin on the expression of apelin and its receptor of the two-kidney, one-clip (2K1C) rats.</p><p><b>METHOD</b>Tirty male Sprague-Dawley rats were randomly divided into normal control group (C), model group (M) and puerarin group (P). The mean of carotid arterial pressure (mCAP), mean of left ventricular end diastolic pressure (LVEDP), and the weight ratio of left ventricular mass (left ventricle plus septum) to bodyweight (LVM/BW) were measured to evaluate the model of 2K1C renal hypertension. The concentrations of apelin in the plasma and left ventricle (LV) were measured with radioimmunoassay. Apelin mRNA and APJ mRNA expressed in the LV were examined by reverse transcription-polymerase chain reaction (RT-PCR). The peptides of apelin and APJ expressed in the LV were detected with immunohistochemistry (IHC).</p><p><b>RESULT</b>Compared with C group, the mCAP, LVEDP and LVM/BW of M group were higher 36.58%, 333.8% and 20.24%, respectively (P<0.05, P<0.01, P<0.01). Compared with M group, LVEDP and LVM/BW of P group were lower 65.24% and 13.12%, respectively (both P<0.05). However mCAP was of no significant difference between these two groups. The levels of apelin-36 in the plasma and LV of M group were respectively higher 18.56% and 207.38% than those of C group (both P<0.05), while ones of P group were lower 24.21% and 49.40% than those of M group (both P<0.05). The expressions of apelin mRNA and APJ mRNA at left ventricle tissues of 2K1C rats were higher 77.66% and 119.00% (both P<0.05) than those of C group. The ones of P group were lower 27.40% and 45.66% than those of M group (both P<0.01). The IHC results indicate that the expressions of apelin and APJ peptides at left ventricle tissues of 2K1C rats were higher 129.51% and 154.1% (both P<0.01) than those of C group, respectively. Whereas the ones of P group were lower 65.36% and 62.87% than those of M group (both P<0.01).</p><p><b>CONCLUSION</b>Through regulating apelin/APJ system puerarin has protective effect on the development of left ventricular hypertrophy by renal hypertension.</p>
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Animais , Masculino , Ratos , Apelina , Receptores de Apelina , Proteínas de Transporte , Genética , Metabolismo , Expressão Gênica , Hipertensão Renal , Tratamento Farmacológico , Metabolismo , Hipertrofia Ventricular Esquerda , Imuno-Histoquímica , Peptídeos e Proteínas de Sinalização Intercelular , Isoflavonas , Usos Terapêuticos , Radioimunoensaio , Ratos Sprague-Dawley , Receptores Acoplados a Proteínas G , Genética , Metabolismo , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
The functional experiment teaching is an inspection to students of their thinking methods and the operation abilities,and is also the summary and sublimation of this course theories teaching.To adapt to the demand for cultivation of the talented person by innovation education,we probe into the improvement of teaching methods of the functional design experiment,and practiced the new teaching system of "six processes"teaching methods such as speaking briefly,giving a demonstration,questioning,giving clue,study and discussion,evaluation,and push forward the biomedical science modes toward the development of "living creature-mental state-social medical science mode"direction.
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To study the stimulation of the genioglossus with percutaneous biphasic current pulses as a new therapeutical method to treat the obstructive sleep apnea syndrome (OSAS), polysomnography (PSG) was used to synchronously monitor the patient. When OSAS was occurring, the stimulation with the optimal parameter was given in time to make the tongue move forward, the glossopharyngeal airway dilated, the resistance of the upper respiratory tract reduced, the hypoxia at night to be improved and the sleeping structure to be ameliorated because of the function of the dilated muscle of the upper airway. The results of the clinical therapeutic effect indicated that 17 of 22 patients with OSAS had cured effects, 2 of whom improved and 3 of whom were without effect. The effective rate was 77.27%. It is preliminarily proved that this is a new method in the treatment of patients suffered from OSAS.
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Adulto , Idoso , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Terapia por Estimulação Elétrica , Métodos , Apneia Obstrutiva do Sono , Terapêutica , Resultado do TratamentoRESUMO
Aim To explore the effects of Puerarin on the Apelin and its receptor APJ of lung tissue in rats with hypoxia pulmonary hypertension.Methods Thirty SD rats were randomly divided into normal control group(N),and hypoxia hypercapnia group(F),and hypoxia hypercapnia+Puerarin group(P).The levels of Apelin-36 in serum and in lung tissue were measured by radioimmunity,the expressions of Apelin and APJ in pulmonary tissuse were observed by RT-PCR.Results ①Mean pulmonary arterial pressure(mPAP),weight ratio of RV to LV+S,the levels of Apelin in serum and in lung tissue of group F were significantly higher than those of group N(P
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Objective To investigate the probable function of new peptide apelin in pathological proceeding of pulmonary hypertension induced by hypoxia.Methods Twenty male Sprague-Dawley rats were randomly divided into normal control group and hypoxia group(9%-11% O_(2),9 h/d,6 d).The mean of pulmonary arterial pressure(mPAP),mean carotid arterial pressure(mCAP) and the weight ratio of right ventricle to left ventricle plus septum(RV/LV+S) were measured.The concentration of apelin in the plasma,right ventricular myocardium and lung tissue were measured by radioimmunity.The expression of apelin gene in right ventricular myocardium and lung tissue were measured by reverse transcription-polymerase chain raction(RT-PCR).Results The mPAP and RV/(LV+S) of hypoxic group were respectively more by 26.06% and 10.83% than control group(P
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AIM:To study lipopolysaccharide (LPS)-stimulated secretion of endothelin-1 (ET-1) and adrenomedullin (Adm) from human vascular endothelial cells (HVEC) and its mechanism. METHODS:In cultured HVEC, LPS was used to stimulate ET-1 and Adm secretion from HVEC. The contents of ET-1 and Adm in medium were determined by radioimmunoassay. RESULTS:LPS stimulated secretion of ET-1 and Adm from HVEC in time-dependent and concentration-dependent manner. The ratio of secreted ET-1 to Adm was not changed compared with the control group. The increase of ET-1 could be inhibited by inhibitor of extracellular signal-regulated protein kinases (PD098059) and inhibitor of P38 kinase (SB202190)(P<0.01), while the increase of Adm could only be inhibited by SB202190(P<0.05), both had no response to inhibitor of protein kinase C (H7), inhibitor of calmodulin (W7), inhibitor of calcineurin (cyclosporin A) and inhibitor of Ca2+ (nicardipine)(P>0.05).CONCLUSION:ERKs and P38 signal pathways may play an important role in the secretion of ET-1 from LPS -stimulated HVEC, while only P38 kinase signal pathway is invovled in the secretion of Adm.
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AIM:To investigate rat Urotensin-II(ra t U-II)-induced vasoconstriction of rat main pulmonary arteries and the role of mitogen-activated protein kinase(MAPK). METHODS: The main pulmon ary artery was dissected from the male Sprague -Dawley rats and artery ring width was 3-4 mm. Concentration-response curves wer e gene rated to rat U-II(0 03 nmol/L-30 nmol/L).Inhibitor of MAPK,PD 98059(0 1 ?mol/ L -10 ?mol/L) were added into the medium after rat U-II(30 nmol/L)induced vasoc onstriction had reached plateau to construct the relaxant concentration-respons e curves and their EC 50 and E max . RESULTS: Rat U-II was a potent vasoconstrictor of isolated rat main pulmonary arteries [EC 50 =7 95?0 4 0, E max =(14 28?6 34)% of the response to 60 mmol/L KCl]; PD 98059 caused c oncentration-dependent relaxations of rat U-II precontracted arteries [EC 50 =5 91?0 45, E max =(81 39?13 65)%]. CONCLUSION: Rat U- II was a potent vasoconstrictor of rat main pulmonary arteries and this response was med iated through MAPK.
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AIM: To investigate the roles of nitric oxide/L-arginine(NO/L-Arg) pathway and urotensin-Ⅱ(UⅡ) in the development of pulmonary hypertension induced by chronic hypoxia-hypercapnia in rats.METHODS: Forty male Sprague-Dawley rats were randomly divided into four groups(n=10): normal control group(A),hypoxia-hypercapnia+saline group(B),hypoxia-hypercapnia+L-Arg liposome group(C) and hypoxia-hypercapnia+N-nitro-L-arginine methyl ester(L-NAME) group(D).Contents of UⅡ,UⅡ mRNA and receptor of UⅡ(UT) mRNA in pulmonary arterioles were measured with immunohistochemistry analysis and in situ hybridization,respectively.Change of small pulmonary vascular microstructure was also investigated.RESULTS:(1) The mean pulmonary artery pressure(mPAP) and the weight ratio of right ventricle to left ventricle plus septum [RV/(LV+S)] in B and D groups were all higher than those in A group(respectively,P
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AIM:To investigate the effect of puerarin on pulmonary vessel collagen metabolism in pulmonary hypertension rats induced by chronic hypoxia and hypercapnia. METHODS: Collagen Ⅰ,Ⅲ and their mRNA were observed in pulmonary arterioles by the technique of immunohistochemistry and in situ hybridization. RESULTS: ① Light microscopy showed media thickness of pulmonary arterioles was much higher in HH(hypoxic-hypercapnia) group than that of NC(normal control) group, and, vessel cavity turned more straiter in HH group than that of NC group.However, the damage of pulmonary arterioles in HP(hypoxic-pueratin) group was much slighter than that of HH group. ② The levels of plasma ET-1 and lung homogenates Hyr were much higher in HH group than those of NC group( P 0.05). CONCLUSION: Puerarin inhibited the deposition of collagen and improved pulmonary vessel remodeling.
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AIM: To study the effect of chronic hypoxia on L-Arginine/NO pathway in rat pulmonary artery. METHODS: Changes in pulmonary artery L-Arginine(L-Arg) transport, nitric oxide synthase (NOS) activity, plasma nitrite level and L-Arg level in HPH rats were investigated. RESULTS: (1) The mean pulmonary arterial pressure (mPAP) and weight ratio of right ventricle to left ventricle and septum (RV/LV+S) of HPH group were higher than those in control group (P
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AIM: To investigate the expression of matrix metalloproteinases(MMPs) in pulmonary arterioles of rats with chronic hypoxia and hypercapnia-induced pulmonary hypertension. METHODS: MMP-2, MMP-9 and MMP-2 mRNA, MMP-9 mRNA were observed in pulmonary arterioles by the techniques of immunohistochemistry and in situ hybridization. RESULTS: ①The mean pulmonary artery pressure (mPAP) and weight ratio of right ventricle to left ventricle and septum (RV/LV+S) of hypoxia-hypercapnia groups were higher than those of normal control group ( P
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AIM: To investigate the effect of chronic hypoxia-hypercapnia and L-arginine (L-Arg) liposome on L-Arg transport in rats pulmonary artery. METHODS: Forty Sprague-Dawley rats were randomly divided into four groups, normal control group (NC), chronic hypoxia-hypercapnia group (HH), chronic hypoxia- hypercapnia group+L-Arg (HL) and chronic hypoxia-hypercapnia group+L-Arg liposome (HP). Changes in pulmonary artery L-Arg transport and pulmonary arterial microscopy were observed. RESULTS: (1) The mean pulmonary artery pressure (mPAP) and weight ratio of right ventricle to left ventricle and septum (RV/LV+S) in HH group were higher than those in NC group, and in HP group was lower than that in HH group and HL group, but there was no significant difference between HL group and HH group; (2) At 0.005 mmol/L, 0.01mmol/L, 0.02mmol/L, 0.05 mmol/L, 0.1 mmol/L and 0.2mmol/L concentration of L-Arg, the velocity of L-Arg transport in HH group was lower than that in NC group, and in HL group higher than in HH group, and in HP group was much higher than that in HH group and in HL group. (3) Light microscopy showed that vessel well area/total area (WA/TA) and media thickness of pulmonary arterioles (PAMT) were much higher in rats of HH group than those in NC group, WA/TA and PAMT in HP group were obviously improved. CONCLUSION: The above results indicated that there existed a functional disturbance in L-Arg transport of pulmonary artery in rats chronically exposed to hypoxia-hypercapnia, and it was obviously enhanced when liposome was used as L-Arg carrier. Thus, it appears that liposome-L-Arg may have clinical perspective in the treatment of chronic hypoxic pulmonary hypertension.
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AIM: To study lipopolysaccharide (LPS)-stimulated secretion of endothelin-1 (ET-1) and adrenomedullin (Adm) from human vascular endothelial cells (HVEC) and its mechanism. METHODS: In cultured HVEC, LPS was used to stimulate ET-1 and Adm secretion from HVEC. The contents of ET-1 and Adm in medium were determined by radioimmunoassay. RESULTS: LPS stimulated secretion of ET-1 and Adm from HVEC in time-dependent and concentration-dependent manner. The ratio of secreted ET-1 to Adm was not changed compared with the control group. The increase of ET-1 could be inhibited by inhibitor of extracellular signal-regulated protein kinases (PD 098059 ) and inhibitor of P38 kinase (SB 202190 )( P 0.05).CONCLUSION: ERKs and P38 signal pathways may play an important role in the secretion of ET-1 from LPS -stimulated HVEC, while only P38 kinase signal pathway is invovled in the secretion of Adm.
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AIM:To observe the alterations in cognition of growing rats exposed to chronic intermittent hypoxia (CIH) and to explore its underlying mechanisms. METHODS:Forty male Sprague-Dawley rats (3-week-old~4-week-old and 80 g to 100 g),which had been trained to complete the 8-arm (4-arm baited) radial maze,were randomly divided into 4 groups:2-weeek-CIH group (2IH),4-week-CIH group (4IH),-week -control group (2C) and 4-weeek-control group (4C). The intermittent hypoxia model was induced by putting the animals in an intermittent hypoxia cabin. When intermittent hypoxia was terminated,spatial memory of these growing rats was tested by 8-arm (4-arm baited) radial maze task,then,one rat in each group was randomly selected for ultrastructural observation. The hippocampus and prefrontal cortexes of the rats were collected for analyzing the mRNA and protein expression of CREB by RT-PCR and Western blotting,respectively. RESULTS:(1) In the 8-arm (4-arm baited) radial maze task,the results indicated that the rats in the 4 groups displayed significant difference in their performance assessed by three measuremens:the reference memory error,the working memory error and total memory error (P 0. 05,respectively). CONCLUSION:Exposure to experimentally-induced IH in growing rats is associated with time related spatial memory impairment. Chronic intermittent hypoxia leads to the disorders of neuron ultra-structure in memory related brain regions. It also inhabits the CREB transduction,expression and CREB phosphorylation,decreases the synthesis of the memory related protein. These factors maybe contribute to learningmemory impairment of growing rats exposed to chronic intermittent hypoxia.