Alteration of myocardial sarcoplasmic reticulum Ca2+ transport protein expression in neonatal hypothyroid rats

AIM: To investigate the alteration of sarcoplasmic reticulum (SR) Ca²⁺ transport proteins including sarcoplasmic reticulum Ca²⁺-ATPase 2a(SERCA2a) and phospholamban(PLB) mRNA expression as well as the alteration of myocardial SR Ca²⁺-ATPase activity in neonatal hypothyroid rats, and to explore the effect of levothyroxine(L-T₄) substitution therapy on the above indexes.METHODS: Hypothyroidism was induced by the administration of propylthiouracil (PTU, 50 mg/d) to the pregnant SD rats by gavage beginning on embryonic day 15 and continuing throughout the lactational period. A subgroup of neonatal hypothyroid rats were intraperitoneally injected with L-T₄ levothroxine (20 μg/kg BW daily), starting from the day of birth. Other pregnant SD rats received normal saline instead of PTU. The samples of the rats in all 3 groups were harvested at postnatal day 3, 5 and 7 respectively (n=10). After measurement of serum thyroid hormone levels, the hearts were removed and the ventricles were weighed (HW). The concentration of calcium in ventricular myocardium(ventricular myoCa²⁺) was detected by fluorospectrophotometry and the activity of SR Ca²⁺-ATPase was determined by the inorganic phosphorus method. The mRNA expression of SERCA2a and PLB was also detected by real-time PCR. RESULTS: Neonatal hypothyroid rats had a significant lower level of SERCA2a mRNA (P<0.05) and a higher level of PLB mRNA (P<0.01), and subsequent lower SERCA2a/PLB at each postnatal day (P<0.01) was observed. Compared with hypothyroid group, the mRNA expression of SERCA2a significantly increased (P<0.05) and that of PLB significantly decreased (P<0.05) in L-T₄ treatment group. The concentration of ventricular MyoCa²⁺ in hypothyroid group was significantly higher than that in control group (P<0.01), and that in L-T₄ treatment group showed a significant decrease as compared with hypothyroid group (P<0.05). The activity of sarcoplasmic reticulum Ca²⁺-ATPase in hypothyroid group was significantly lower than that in control group (P<0.01), and that in L-T₄ treatment group showed a significant increase as compared to hypothyroid group (P<0.05). CONCLUSION: The deficiency of thyroid hormone, resulting in decreased expression of SERCA2a mRNA as well as increased PLB mRNA, contributes to the reduction of SR Ca²⁺-ATPase activity in neonatal rats. This may be one of the most important mechanisms of myocardial systolic and diastolic dysfunctions.     

Effects of urocortin-I preconditioning on myocardial mitochondrial respiratory function and enzyme activity in rats

ATM: To investigate the influence of urocortin-I (Ucn I) preconditioning on the myocardial mitochondrial respiratory function and enzyme activity in the rats with ischemia reperfusion, and to observe the changes of ATP content in the myocardial cells. METHODS: (1) The healthy male Sprague-Dawley rats were randomly divided into 4 groups:normal group (Nor group), ischemia reperfusion group (IR group), Ucn I preconditioning group (Ucn I group), 5-hydroxy acid (5-HD)+Ucn I group. Langendorff perfusion was used to establish the in vitro model of cardiac ischemia reperfusion. At the end of the balance (T₁), before ischemia (T₂) and at the end of the reperfusion (T₃) respectively, the myocardial mitochondria was extracted, the mitochondrial respiratory function and respiratory enzyme activity in each group were determined. (2) The method of MPA isolated heart perfusion was used to isolate myocardial cells of the adult rats. After cultured for 24 h, myocardial cells were divided into 4 groups:Nor group, hypoxia/reoxygenation group (I/R group), Ucn I group, 5-HD+Ucn I group. Hypoxia/reoxygenation model of myocardial cells was established. At the end of reoxygenation, the changes of myocardial ATP content were measured by high performance liquid chromatography.RESULTS: (1) Compared with T₁, T₂ time points, the respiratory function (state 3 respiratory rate, respiratory control rate) and NADH oxidase, succinate oxidase and cytochrome C oxidase activities at T₃ time point were significantly decreased (P<0.05) in all groups except Nor group. At T₃ time point, the myocardial mitochondrial respiratory function and respiratory enzyme activity in Ucn I group were superior to 5-HD+Ucn I group and IR group (P<0.05), but was inferior to Nor group (P<0.05). At T₃ time point, the respiratory function of myocardial mitochondria and respiratory enzyme activities (NADH oxidase, succinate oxidase) in 5-HD+Ucn I group were better than those in IR group (P<0.05), but no statistical difference of the cytochrome C oxidase activity between the 2 groups was observed. The respiratory function and 3 kinds of respiratory enzyme activities at T₁, T₂ time points had no statistical change. (2) At the end of the reoxygenation, the myocardial ATP content in Nor group was higher than that in other groups (P<0.01). The myocardial ATP contents in I/R group and 5-HD+Ucn I group were lower than that in Ucn I group (P<0.05). In additon, 5-HD+Ucn I group was higher ATP content compared with I/R group (P<0.05). CONCLUSION: Ucn I preconditioning attenuates the ischemia/reperfusion induced damages of myocardial mitochondrial respiratory function and respiratory enzyme activity, thus ensuring the myocardial ATP contents under the condition of hypoxia/reoxygenation.     

Role of endogenous arginine vasopressin in soman-induced hypothermia in rats

AIM: This study was designed to examine whether endogenous arginine vasopressin (AVP) is involved in soman-induced hypothermic process. METHODS: Core temperature was measured at 60 min intervals with digital thermometer. Effect of AVP V₁ receptor antagonist (30 μg/kg, ip) on soman-induced hypothermia was observed in rats, and plasma AVP concentration was measured at 2 h after administration of soman(60 μg/kg, sc). RESULTS: Administration of soman led to a marked hypothermia. Core temperature recovered to basal levels at 7 h after soman treatment. Plasma AVP concentration increased markedly at 2 h after soman treatment, and administration of AVP V₁ receptor antagonist markedly blocked the hypothermic effect of soman. CONCLUSION: The data indicate that AVP is involved in soman-induced hypothermia in the rat.     

Effects of remifentanil on monophasic action potential and transmural dispersion of repolarization in rabbit myocardium

AIM: To study the effect of remifentanil on monophasic action potential and transmural dispersion of repolarization (TDR) in the 3-layer myocardium of isolated rabbit hearts. METHODS: Adult rabbits (n=18, 2.0 ~ 2.5 kg) were used to isolate the hearts for preparing Langendorff perfusion model. The hearts were randomly divided into 3 groups after perfusion with K-H solution for 15 min: the perfusion in control group (C group) continued for 60 min; the hearts in remifentanil group (R group) were perfused with 12 μg/L remifentanil K-H solution for 60 min; the hearts in remifentanil+aminophylline group (RA group) were given 60-min perfusion of 12 μg/L K-H remifentanil+30 mg/L aminophylline. The HR and 3 layers of myocardial monophasic action potential (MAP) in the left ventricular anterior wall were recorded at time points after balanced infusion for 15 min (T₀), and continued perfusion for 15 min (T₁), 30 min (T₂) and 60 min (T₃). The monophasic action potential duration of repolarization at 90% (MAPD₉₀) and the transmural dispersion of repolarization (TDR) were calculated. The early afterdepolarization, delay afterdepolarization and arrhythmia were also observed. RESULTS: In R group, slower HR and prolonger MAPD₉₀ and TDR at T₁~T₃ were observed as compared with those at T₀ (P<0.05). R group showed slower HR and longer MAPD₉₀ and TDR than C group and RA group (P<0.05). CONCLUSION: Remifentanil slows the HR, extends the MAPD₉₀ and increases the TDR, thus being prone to induce reentry. Aminophylline makes HR faster and MAPD₉₀ shorter, thereby reducing the TDR.     

Effects of hypoxia and hypercapnia on expression of soluble guanylate cyclase in rat pulmonary artery

AIM:To investigate the expression of soluble guanylate cyclase protein and its mRNA in rat pulmonary artery after exposure to hypoxia and hypercapnia.METHODS:Male Sprague-Dawley rats were randomly split into 4 group, which were hypoxic hypercapnic (HH 1 week, HH 2 weeks, HH 4 weeks) group and control group, to copy pulmonary hypertensive animal model. The expression of sGCα₁ and β₁ subunits protein of medial and small pulmonary artery was performed by immunohistochemistry with a polycolonal antibody. In situ hybridization was performed on the rat lung tissue using sGC oligonuclear probe to assay the expression of sGCα₁subunit mRNA.RESULTS:The sGCα₁ and β₁ subunits protein and sGCα₁ subunit mRNA were faint staining in the pulmonary small and medium artery in HH1 week, HH 2 weeks and HH 4 weeks groups compared with control group (all P＜0.01).CONCLUSION:sGC subunit mRNA and protein expression in pulmonary small and medium artery were decreased after exposure to hypoxia and hypercapnia, which took part in the development of the pulmonary hypertension.     

Age-associated changes of p38 MAPK and JNK signal pathways in cardiac fibroblasts treated with transforming growth factor beta 1

AIM:To investigate the effect of aging on p38 mitogen-activated protein kinase(MAPK) and c-Jun N-terminal kinase(JNK) signal pathways in rat cardiac fibroblasts(CFs). METHODS:Cardiac fibroblasts obtained from neonatal and aged rats were cultured and randomly divided into 4 groups:neonatal PBS control group(N1 group), neonatal TGF-β₁ treatment group(N2 group), aged PBS control group(A1 group) and aged TGF-β₁ treatment group(A2 group). Proliferation of CFs was detected by MTT coloricmetric assay. The expression levels of total p38 MAPK, JNK, phospho-p38 and phospho-JNK were measured by Western blotting. RESULTS:The proliferative capacity of aged CFs was significantly decreased as compared with neonatal CFs after stimulated with TGF-β₁. In response to TGF-β₁, the expression levels of phospho-p38 and phospho-JNK were significantly increased in N2 group and A2 group as compared with N1 group and A1 group, respectively. The levels of total p38 and nonphosphorylated JNK in N2 group were similar to those in A2 group. Compared with N2 group, the levels of phospho-p38 and phospho-JNK markedly decreased in A2 group. CONCLUSION:These data indicate that p38 MAPK and JNK signal pathways are impaired in aged CFs.     

黄瓜种质资源苗期耐低温性评价

以43份不同类型的黄瓜品种(品系)为试验材料,在黄瓜苗期进行低温处理(昼/夜温度为12℃/9℃),以冷害指数作为鉴定指标对其耐低温性进行评价。最终鉴定出7份耐低温材料、10份低温敏感型材料。对黄瓜低温处理后的冷害指数(Y)与叶绿素降低率(X1)、可溶性蛋白增高率(X2)、MDA增高率(X3)、脯氨酸增高率(X4)、POD增高率(X5)进行相关性分析,并建立了鉴定黄瓜耐低温性的回归方程:Y=0.433+0.671X1+0.005X2+0.121X3-0.733X4+0.248X5。     

Dynamical observation of the expression of TGF-β1 and MAPK1/3 in the renal tubules of rats with diabetes

AIM: To observe the expression of transforming growth factor β₁ (TGF-β₁), MAPK_1/3 and fibronectin (FN) in the development of renal tubulointerstitial disease. METHODS: Wistar male rats were randomly divided into normal control group, diabetic group of 1week, 2 weeks, 4 weeks and 8 weeks. Diabetic model was induced by peritoneal injection of streptozotocin. Immunohistochemistry was employed to detect the expression of TGF-β₁, MAPK_1/3 and FN in the kidney. TGF-β₁ protein in the renal cortex was checked by Western blot. BG, Scr and UP were analysed by biochemical methods, and the morphological changes in renal tubulointerstitium were also examined under microscopy on sections stained with HE and PAS. RESULTS: The expression of MAPK_1/3 and FN was observed, but not the expression of TGF-β₁ in normal renal tissue. Positive staining of TGF-β₁ was observed in the renal tubulo-interstitium in 1-week diabetic group and thereafter it increased in the course of diabetes. A continuous increase in the expression of MAPK_1/3 and FN was also observed in two - week diabetic rats. Chronologically the expression of TGF-β₁,MAPK_1/3 and FN and the ratio of KW/BW were positively correlative with each other in diabetic animals except one -week diabetic rats. There was also a positive correlation between MAPK_1/3 and FN in l -week diabetic rats. CONCLUSION: Our data suggest that TGF-β₁ appears in the renal tubulointerstitium in early period of diabetes and then its signal is mediated by MAPK_1/3 cascades to accelerate production of FN ,and in turn leads to renal hypertrophy and tubulointerstitial fibrosis.     

Influence of losartan potassium on renal expression of TGF-β1, CD68 and MCP-1 in type 2 diabetic nephropathy rats

AIM: To observe the effects of losartan potassium on renal expression of transforming growth factor beta 1(TGF-β₁), CD68 and monocyte chemoattractant protein-1 (MCP-1) in type 2 diabetic nephropathy rats for exploring the protective mechanism of losartan potassium on type 2 diabetic rat kidney. METHODS: Thirty Sprague-Dawley rats were randomized into 3 groups: normal control group, model group and treatment group. The morphology of kidney tissues, the renal function, and the change of 24 h urinary protein quantitative index were measured after 15 weeks of treatment, while TGF-β₁, CD68 and MCP-1 expression in kidney cortex was observed by immunohistochemistry. RESULTS: Compared with the normal control rats, the body weight of the rats was lower in other groups, but the levels of blood glucose, triglyceride and cholesterol were higher.The expression of CD68, MCP-1 and TGF-β₁, 24 h urinary protein quantitative index and serum creatinine were higher in model group than those in normal control rats. However, compared with model group, serum creatinine, 24 h urinary protein quantitative index and the expression of CD68, MCP-1 and TGF-β₁ were decreased in treatment group. CONCLUSION: Losartan potassium protects the kidney of diabetic nephropathy rats through inhibiting the expression of TGF-β₁ and MCP-1 in the kidney and restraining macrophage infiltration.     

Role of bradykinin in the therapeutic effect of enalapril on left ventricular hypertrophy and cardiac function after myocardial infarction in rats

AIM:To investigate the influences of bradykinin(BK)on left ventricular hypertrophy and cardiac function in angiotensin-converting enzyme inhibitor(ACEI) therapy in rats after myocardial infarction.METHODS:The effects of enalapril (500 μg·kg^-1·d^-1), enalapril (500 μg·kg^-1·d^-1)with BKB₂ receptor antagonist (Hoe-140 500 μg·kg^-1·d^-1), losartan(3 mg·kg^-1·d^-1) on left ventricular end-diastolic pressure (LVEDP), maximum positive left ventricular pressure change (+dp/dt_max) and LVW/BW as well as V(m)n of noinfarcted area were examined after 4 weeks treatment in rats after myocardial infarction.RESULTS:The values of LVEDP, LVW/BW and V(m)n of three treatment groups were higher than that of untreated MI group (P＜0.05),but the +dp/dt_max of three treatment groups were not significantly different compared with the untreated MI group. In addition, no significant difference in MAP was observed among the three treatment groups, but the LVW/BW and V(m)n of enalapril+Hoe-140-treated group were higher than that of enalapril-treated group (P＜0.05) .CONCLUSION:Enalapril can prevent left ventricular hypertrophy and improve cardiac function independent of blood pressure after myocardial infarction, which is partly due to the inhibition of BK degradation.     

Protective effect of magnesium on brain with acute ischemia and reperfusion injury

AIM: To dynamically observe the protective effect of magnesium on the brain with ischemia and reperfusion injury in the middle cerebral artery occlusion (MCAO) rat model using the device of laser Doppler flowmetry (LDF) .METHODS: Twenty-four Sprague-Dawley male rats (280-300 g) were used to establish MCAO model with the conventional line-embolism method.The rats were divided into 4 groups according to the initial time of peritoneal injection of magnesium sulfate (MgSO₄).The rats in MgSO₄ groups were treated with 25% MgSO₄ at 160 mg/kg at time points of 20 min, 30 min and 40 min after ischemia.The rats in control group were treated with the same volume of normal saline.The real-time fluctuation of the local cerebral blood flow (CBF) was dynamically monitored by LDF.The degree of nervous functional defect was evaluated by calculating the neurological impairment score of the rats after suffered from 2 h ischemia and followed with 24 h reperfusion.All the rats were killed with an end breaking method, and the volumes of brain infarction were evaluated by TTC staining at the brain slices obtained by autopsy.RESULTS: The fluctuating features in 3 MgSO₄ treatment groups were as follows: when reperfusion began, the local CBF appeared and increased smoothly, then approached to its baseline step by step, and kept at the stable level in the whole reperfusion period.The CBF in control group fluctuated sharply in the whole reperfusion period associated with the heart beating, which was significantly different from those in MgSO₄ treatment groups.The average volume of brain infarction in MgSO₄ treatment groups was 26.5%, 36.5% and 24.5%, respectively, and was 54.0% in control group.The local nervous defect scores in MgSO₄ treatment groups were 5.670±1.003, 8.670±1.211 and 7.170±1.472, respectively, and was 11.170±0.983 in control group.CONCLUSION: Vasomotor functional improvement might be one of the protective mechanisms of magnesium on the brain with cerebral ischemia and reperfusion injury in acute ischemic stroke.     

Effects of thyroid hormone on the myosin heavy chain mRNA expression in cardiac myocytes induced by angiotensin Ⅱ

AIM: To study the effect of thyroid hormone on the expressional change of myosin heavy chain(MHC) gene in cardiomyocyte induced by angiotensinⅡ(AngⅡ) and its potential mechanism. METHODS: Cardiac myocyte was cultured according to the method of Simpson. 10^-8 mol/L T₃ and 10^-7 mol/L AngⅡ were added to the culture medium, respectively or synchronously. After 48 h, the expression of α and β-MHC mRNA in myocytes were detected by RT-PCR. The protein kinase C activation were detected by PepTag non-radioactive PKC assay. The incorporation of -Leucine and [³H]-thymine to test the protein and DNA synthesis in myocytes were also performed. RESULTS: AngⅡalone increased the incorporation of [³H]-Leucine of myocytes while it had no effect on the incorporation of [³H]-thy mine. The expression of β-MHC mRNA was increased and the expression of α-MHC mRNA was decreased significantly at the condition of AngⅡ. The enhanced PKC activation was induced by AngⅡalso. When AngⅡand T₃ were added to the culture medium synchronously, though the incorporation of [³H]-leucine and [³H]-thymine were not changed compared with AngⅡ treated alone. The α-MHC mRNA expression was increased and the β-MHC mRNA expression was decreased significantly. The PKC activation of the myocytes also was decreased. CONCLUSIONS: T₃ inhibited the expressional change of myosin heavy chain gene in cardiac myocytes induced by AngⅡ. The effect of T₃ on the change of PKC activation in cardiac myocytes may be one of its mechanisms.     

Effects of drynaria total flavonoids on serum levels of leptin,interleukin 6, prostaglandin E2 and expression of bone β2-adrenergic receptor in ovariectomized osteoporosis rats

AIM: To investigate the effects of drynaria total flavonoids on serum levels of leptin (LEP), interleukin 6(IL-6), prostaglandin E₂(PGE₂) and the expression of bone β₂-adrenergic receptor (ADRB2) in a rat model of ovariectomized osteoporosis(OP). METHODS: The osteoporosis model was established by ovariectomy. Twelve weeks after modeling,bone mineral density (BMD) was determined by dual-energy X-ray absorptiometry to verify successful modeling.Enzyme-linked immunosorbent assay was applied to detect the concentrations of LEP, IL-6 and PGE₂ in serum. The expression of ADRB2 was determined by immunohistochemical technique. RESULTS: Compared with sham group,BMD of the rats in model group significantly decreased in multiple regions 12 weeks after modeling(P<0.01). The serum levels of LEP, IL-6 and PGE₂ in model group were significantly higher than those in sham group(P<0.05). The levels of LEP, IL-6 and PGE₂ in drynaria total flavonoids group were significantly lower than those in model group(P<0.01). No significant difference of PGE₂ between these 2 groups was observed. The ADRB2 expression in sham group and treatment group was significantly different from that in model group, and no significant difference between sham group and treatment group was found. CONCLUSION: The serum levels of LEP, IL-6 and PGE₂ and the expression of bone ADRB2 increased in OP rats.Drynaria total flavonoids reduce the production of LEP, IL-6 and the expression of ADRB2, and suppress the bone absorption, which may be one of the mechanisms in treating OP.     

Effects of icariin on expression of transforming growth factor β1 and type Ⅳ collagen in diabetic rat testis

AIM: To determine the beneficial effects of icariin on streptozotocin (STZ)-induced diabetic testopathy in rats. METHODS: The diabetic animal model was induced in male Sprague-Dawley rats by an injection of streptozotocin (40 mg/kg, iv). The rats were randomly divided into 3 groups: control group, model group and icariin (80 mg/kg, ig) group. Twelve weeks after injected with streptozotocin, all rats were anaesthetized and killed to remove the testes from scrotum. Serum concentrations of glucose and testosterone, and the levels of succinate dehydrogenase (SDH), acid phosphatase (ACP), γ-glutamyl transpeptidase (γ-GT) and lactate dehydrogenase (LDH) in testes were measured. The morphology of the testicular tissues was observed under light microscope. Immunohistochemistry was employed to determine the protein levels of TGF-β₁ and type Ⅳ collagen. RESULTS: Compared with control group, the content of serum glucose increased while the serum level of testosterone and the activitiy of SDH, ACP, γ-GT and LDH in testis decreased in model group (P<0.01). The histopathological examination showed that the diameters of seminiferous tubules and various grades of spermatocytes in the testis were markedly decreased. Compared with control group, the expression of TGF-β₁ and collagen Ⅳ was significantly increased in model group. These alterations were significantly attenuated in icariin group (P<0.01). CONCLUSION: Icariin evidently relieves testicular damage in rats with diabetic testopathy by improving the secretion of testosterone and reducing the expression of TGF-β₁ and collagen Ⅳ at protein level.     

Influence of exogenous somatostatin on pancreatic blood flow in normal rats or rats with acute necrotizing pancreatitis

AIM:To investigate the influence of exogenous somatostatin (stilamin) on pancreatic blood flow in normal rats or rats with acute necrotizing pancreatitis (ANP).METHODS:Pancreatic blood flow (PBF) was detected with computerized tissue blood flowmeter and rats with ANP were triggered with sodium taurocholate. Metabolites of eicosanoids in plasma were determined with radioimmunoassay. Other laboratory tests including histopathologic observation under optical or electron microscope were used. RESULTS:There was a significant decrease in PBF in normal rats after stilamin administration in comparison with that before use of the drug. There was significant decrease in PBF after onset of ANP, but, compared with that in ANP group, significant increase was shown in SS(stilamin)+ANP group. Plasma thromboxin-B₂(TXB₂) in ANP group at 6 hours after ANP was significantly higher, with increase of 4.5 times, than that in Sham(sham operated) group while TXB₂, detected each time during the course of ANP, significantly decreased in SS+ANP group. 6-Keto-prostagland in F_1α(6-Keto-PGF_1α) at 6 h after ANP was significantly higher, and the ratio of TXB ₂／6-Keto-PGF_1α, significantly lower in SS+ANP group than that in ANP group. Lessened necrosis of acinar cells, along with much fewer microthrombi in microvessels in SS+ANP group, was shown by pathologic scoring or electron microscope than that in ANP group.CONCLUSION:Administration of exogenous somatostatin leads to the decrease in PBF in physiological setting but it attenuates pancreatic ischemia in SS+ANP group, which may be attributed to correction of abnormal metabolism of eicosanoids, improvement of pancreatic microcirculation and cytoprotection of acinar cells as well.     

Effect of tenuigenin on tau protein phosphorylation at Ser396 site in neurons of AD rats induced by Aβ1-40

AIM:To investigate the effect of tenuigenin(TEN) on hyperphosphorylation of tau protein in neurons of amyloid β-peptide_1-40(Aβ_1-40) -induced Alzheimer disease(AD) rats. METHODS:Aβ_1-40 was injected into hippocampus CA1 region of the rats to establish AD model. TEN at different doses(18.5 mg/kg, 37.0 mg/kg and 74.0 mg/kg) was intragastrically administered. The protein expression of protein kinase A(PKA),protein phosphatase 2A(PP2A), total tau and p-tau(Ser³⁹⁶) in the neurons was observed by the method of immunohistochemistry. The protein content of total tau and p-tau(Ser³⁹⁶), and the expression level of PKA and PP-2A were detected by Western blotting analysis. RESULTS:In Aβ_1-40 group, the level of total tau, the phosphorylation of tau protein and the expression of PKA were significantly increased compared with those in sham operation group. Meanwhile, the expression of PP2A in Aβ_1-40 group was lower than that in sham operation group. In TEN treatment group, the level of total tau, the phosphorylation of tau protein and the expression of PKA were markedly decreased, and the expression of PP2A was increased as compared with Aβ_1-40 group. CONCLUSION:TEN may protect the neurons from the toxic effect of Aβ_1-40 and reduce the hyperphosphorylation of tau(Ser³⁹⁶) in the neurons of AD rats by activating the expression of PP2A and inhibiting the expression of PKA.     

Influences of Ca2＋ antagonist on the mRNA levels of L-type Ca2＋ channel and Na＋-Ca2＋ exchanger in the atria of hypertensive rats

AIM: To investigate the mRNA changes of L-type Ca²⁺channel α_1Csubunit (CaL-α_1C) and Na⁺-Ca²⁺ exchanger (NCX) in the atria of renovascular hypertensive rats. METHODS: Two-kidney one-clip Goldblatt hypertensive Sprague-Dawley rats were divided into three groups 1 week after operation, HD group was treated with 250 mg/d diltiazem, LD group was treated with 50 mg/d diltiazem, control group (C group) was treated with vehicle. After 4 weeks treatment, semiquantitative RT-PCR was used to estimate the mRNA changes of CaL-α_1Cand NCX, and GAPDH was used as internal control. RESULTS: Systolic blood pressure in LD group was comparable with C group, and that in HD group was decreased to normal level after diltiazem treatment. In C group, CaL-α_1C mRNA level were 2.5, 2.4 and 2.1 times of S, HD and LD group, and NCX mRNA level were 1.9, 1.6 and 2.1 times of S, HD and LD group. There were no significant difference in the mRNA level of CaL-α_1Cand NCX among S, HD and LD group. CONCLUSION: The mRNA levels of CaL-α_1Cand NCX are upregulated in the atria of hypertensive rats. Ca²⁺ antagonist inhibites their upregulation independent of blood pressure.     

Role of TGF-β1/Smads and ERK expression in vascular remodeling induced by high-salt diet in Wistar rats

AIM: To investigate the role of transforming growth factor β₁ (TGF-β₁)/Smads and extracellular signal-regulated kinase(ERK) expression in vascular remodeling induced by high-salt diet in Wistar rats. METHODS: Wistar rats were randomly divided into 3 groups: normal control group (n=13), high salt (8%) model group and high salt+telmisartan group (n=13). Tail-cuff arterial pressure was determined every 2 weeks. After 24 weeks, the rats in high salt model group were divided into model animals with hypertension group (MH, n=12) and model animals without hypertension group (MN, n=12). The remodeling of aorta and mesenteric artery was observed by HE and Masson staining. In addition, the techniques of immunohistochemistry and real-time PCR were applied to detect the expression of proliferating cell nuclear antigen (PCNA), TGF-β₁, p-Smad2/3, p-ERK1/2 and Smad7 at both protein and mRNA levels. RESULTS: Compared with normal control group, blood pressure in MH group was much higher, and media thickness (MT) and collagen volume fraction (CVF) of arteries in MH and MN groups were higher.The mRNA expression of TGF-β₁, Smad2 and Smad7 in the aorta was significantly increased, and the protein levels of PCNA, p-ERK1/2, TGF-β₁ and p-Smad2/3 in the aorta and mesenteric artery media were elevated, but Smad7 decreased. After telmisartan treatment, MT and CVF were much lower,and the protein levels of PCNA, TGF-β₁, p-Smad2/3 and p-ERK1/2 were significantly reduced, whereas Smad7 was increased. CONCLUSION: The abnormal expression of TGF-β₁/Smads and ERK may be involved in the mechanism of remodeling of aorta and mesenteric artery induced by high-salt diet. Telmisartan prevents the vascular remodeling via regulating TGF-β₁/Smads and ERK signal pathways mediated by angiotensinⅡ type 1 (AT₁) receptor, at least in part.     

Roles of ERK pathway and Panax notoginoside treatment in hypoxic hypercapnia pulmonary hypertension in rats

AIM: To investigate the roles of Panax notoginoside (PNS) and ERK1/2 signaling pathway in the pathological process of chronic hypoxic hypercapnia pulmonary hypertension in rats.METHODS: The animal model of chronic hypoxic hypercapnia pulmonary hypertension was set up in 72 male Sprague-Dawley rats and the animals were randomly divided into 6 groups: normal (N) group, hypoxic hypercapnia for 3-day (H_3d) group, hypoxic hypercapnia for 1-week (H_1w) group, hypoxic hypercapnia for 2-week (H_2w) group, hypoxic hypercapnia for 4-week (H_4w) group and PNS treatment (H_p) group.The rats in H_p group were injected with PNS (50 mg·kg^-1·d^-1, ip) before placing the animals into the hypoxic hypercapnia chamber.The rats in other groups were injected with normal saline (2 mL/kg, ip).The morphological changes of the pulmonary artery were observed under microscope with HE staining.Western blotting was used to detect the protein expression of p-ERK.The protein levels of p-ERK in the lung tissues and pulmonary blood vessels were determined by immunohistochemistry.RESULTS: The ratios of WA/TA in H_1w, H_2w, H_4w and H_p groups were higher than that in N group (P<0.05).The ratio of WA/TA in H_p group was obviously lower than that in H_4w group (P<0.05).The protein expression of p-ERK was barely positive in N group, but was up-regulated in the pulmonary tissues in all hypoxic rats.Compared with N group, the protein level of p-ERK was markedly up-regulated in H_3d group, reached its peak in H_2w group, and tended to decline in H_4w group (P<0.05).In pulmonary arterial tunica intima and tunica media, p-ERK protein was dramatically expressed in all hypoxic rats compared with the control animals (P<0.05).In the lung tissues, the protein level of p-ERK in H_p group was lower than that in H_4w group (P<0.05).In pulmonary arterial tunica intima and tunica media, the protein level of p-ERK in H_p group was lower by 84.86% than that in H_4w group (P<0.05).CONCLUSION: ERK1/2 as a signal transducer may play an important role in the development of hypoxia and hypercapnia induced pulmonary hypertension.PNS inhibits the expression of ERK1/2, thus attenuating the development of pulmonary hypertension and improving pulmonary vascular remodeling.     

Effects of endothelial progenitor cell-conditioned medium on proliferation and differentiation of type Ⅱ alveolar epithelial cells exposed to hyperoxia

AIM: To investigate the effect of hyperoxia exposure on the paracrine function of endothelial progenitor cells (EPCs), and to explore the effects of paracrine factors of EPCs on the proliferation and differentiation of type Ⅱ alveolar epithelial cells (AECⅡ) exposed to hyperoxia. METHODS: Bone marrow-derived mononuclear cells were isolated and cultured in EGM-2MV medium for 7~10 d to obtain and identify EPCs. EPCs were cultured in room air (RA) or 60% O₂. The normoxia EPC-conditioned medium (E-CM-RA) and hyperoxia EPC-conditioned medium (E-CM-O₂) were collected. The levels of VEGF, FGF10, PDGF-BB and EGF in E-CM-RA and E-CM-O₂ were detected by ELISA. AECⅡ from adult rats were isolated, purified and cultured for 2 d, then divided into RA group, O₂ group, O₂+E-CM-RA group and O₂+E-CM-O₂ group. The proliferation of AECⅡ was detected by MTT assay and cell counting. The mRNA expression of SP-C and AQP5 was quantified by real-time PCR. RESULTS: The expression of VEGF and FGF10 in E-CM-O₂ group decreased significantly compared with E-CM-RA group (P<0.01). There were significant differences in AECⅡ viability and number among the 4 groups at 12 h, 24 h, 2 d and 3 d (P<0.01). Compared with RA group, AECⅡ viability and number in O₂ group decreased significantly at 12 h, 24 h, 2 d and 3 d (P<0.05). The AECⅡ viability and number in O₂+E-CM-RA group were significantly higher than those in O₂ group at 12 h, 24 h, 2 d and 3 d (P<0.05). However, no significant difference in AECⅡ viability and number between O₂+E-CM-O₂ group and O₂ group at 12 h, 2 d and 3 d was observed. There were significant differences in the mRNA expression of SP-C and AQP5 in the 4 groups at 24 h, 2 d and 3 d (P<0.01). Compared with RA group, the mRNA expression of SP-C in O₂ group was significantly inhibited (P<0.01), but the mRNA expression of AQP5 was promoted (P<0.01) at 24 h, 2 d and 3 d. Compared with O₂ group, the mRNA level of SP-C in O₂+E-CM-RA group and O₂+E-CM-O₂ group (P<0.05) at 24 h, 2 d and 3 d was increased, and the mRNA expression of AQP5 (P<0.01) at 2 d and 3 d was inhibited.CONCLUSION: EPCs secrete VEGF and FGF10, and hyperoxia impairs this paracrine function. Hyperoxia exposure inhibits AECⅡ proliferation and the mRNA expression of SP-C, but promotes the mRNA expression of AQP5. EPC-conditioned medium improves the proliferation of hyperoxia-exposed AECⅡ, and inhibits the transformation of AECⅡ. Hyperoxia exposure impairs the paracrine function of EPCs, and weakened the effects of E-CM-O₂ on AECⅡ.