Expression and distribution of osteopontin in the development of pulmonary hypertension induced by hypoxia-hypercapnia

AIM: To study the expression and distribution of osteopontin (OPN) in lungs and pulmonary arteries in pulmonary hypertensive rats induced by hypoxia-hypercapnia, and to explore the role of OPN in pathogenesis of pulmonary hypertension. METHODS: Forty-eight male Sprague-Dawley rats (Weight 180 g-220 g) were randomly divided into four groups: normal control group (NC), hypoxic hypercapnia 1-week，2-week and 4-week group (1HH, 2HH and 4HH). The expressions of OPN mRNA and protein in lungs and pulmonary arteries were detected by RT-PCR and immunohistochemistry. ELISA was used to detect the concentration of OPN in lung homogenates. The content of OPN in pulmonary arteries was detected by Western blotting. RESULTS: ① The mean pulmonary artery pressure (mPAP) and weight ratio of right ventricle to left ventricle and septum ［RV/(LV+S)］ in all hypoxic hypercapniac groups were higher than those in normal control group (P<0.01), respectively. Differences of mean carotid artery pressure (mCAP) among these four groups were not significant (P>0.05). ② The expression of OPN mRNA was significantly increased in pulmonary arteries and lung tissues in hypoxic hypercapnic groups compared with normal control group (P<0.01). ③ The result of immunohistochemistry showed that OPN was only detected in bronchus and alveolar epithelium, but not detected in pulmonary arterioles of normal control group. In contrast，OPN expression was evident in pulmonary arterioles of 1HH rats，especially in media. Moreover, the expression of OPN was markedly increased in group 2HH and 4HH. ④ OPN levels in lung homogenates in 1HH, 2HH and 4HH were increased by 69%, 128% and 187% (P<0.01), respectively, compared with control rats. ⑤ Western blotting analysis showed that the contents of OPN were significantly higher in all hypoxic hypercapnic groups than those in NC group (P<0.01).CONCLUSION: The expressions of OPN in pulmonary arteioles and lung are increased in rats with pulmonary hypertension. OPN might play an important role in the pathogenesis of pulmonary hypertension induced by chronic hypoxia and hypercapnia.     

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.     

Dexmedetomidine reduced isoflurane-induced neuroapoptosis by inhibiting activation of p38 and JNK proteins in hippocampus of neonatal rats

AIM：To investigate the effect of dexmedetomidine (Dex) on neuronal apoptosis induced by isoflurane (Iso) and its relationship with the expression of p38 mitogen-activated protein kinase (p38) and c-Jun N-terminal kinase (JNK) proteins in the hippocampus of neonatal rats. METHODS：Forty-eight neonatal SD rats at postnatal day 7 were randomly divided into control group (Con), Dex group, Iso group and Iso combined with Dex (Iso+Dex) group. Rats in Iso and Iso+Dex groups were exposed to 0.75% Iso for 6 h, while rats in Con and Dex groups were exposed to air for 6 h. Rats were intraperitoneally injected with 25 μg·kg-1 Dex (Dex and Iso+Dex groups) or 150 μL saline (Con and Iso groups) 20 min before exposure and 2 and 4 h after exposure. After the termination of anesthesia, the neuronal apoptosis in hippocampal CA1 region was detected by TUNEL staining, and the protein expression of cleaved caspase-3, phospho-p38 (p-p38), p38, phospho-JNK (p-JNK) and JNK in hippocampal tissues was detected by Western blotting. RESULTS：The number of TUNEL positive cells in hippocampal CA1 region of the rats in Iso group was increased by 447.57% (P<0.01) compared with Con group, while Dex significantly inhibited the increased TUNEL positive cells in Iso group by 75.18% (P<0.01). The expression of cleaved caspase-3 protein in Iso group was increased by 126.29% (P<0.01) compared with Con group, while Dex reversed the increased cleaved caspase-3 protein expression (P<0.01). Iso significantly increased the phosphorylation of p38 and JNK proteins (P<0.01), while Dex reversed the increased p-p38 and p-JNK proteins (P<0.01). CONCLUSION：Dex attenuates Iso-induced neuroapoptosis in the hippocampus of neonatal rats through inhibiting the phosphorylation of p38 and JNK proteins.     

Effect of JNK expression in dorsal root ganglia on histological change of foot skin in diabetic rats

AIM: To observe the change of skin histology in diabetic rats and to investigate the possible me-chanism of c-Jun N-terminal kinase (JNK) protein in the dorsal root ganglion (DRG) during the process. METHODS: Diabetic animal model was established in the male SD rats by intraperitoneal injection of streptozotocin. Plantar skin specimens of the rats were collected from control group, DM 2-week group (DM2), DM 4-week group (DM4), and DM 8-week group (DM8). Immunohistochemical staining and HE staining were used to observe the change of PGP 9.5 immunoreactive nerve terminals and the structures of the skin tissues. The protein expression of PGP 9.5 in the plantar skin tissues, and JNK and p-JNK protein in the DRG within lumbar 5, 6 (L5, 6), and sacral 1 (S1) spinal cord segments were detected by Western blotting. RESULTS: PGP 9.5 immunoreactive nerve terminals of the plantar skin of the rats mainly distributed in the basal layer of the epidermis and papillary dermis. Compared with control group, PGP 9.5 positive nerve terminals in DM4 group showed reduced density and sparse distribution. PGP 9.5 positive nerve terminals in DM8 group showed significantly reduced distribution, thinner nerve diameter, shorter length and distorted shape. Histological changes of the thinner epidermal tissue, reduced epidermal cell layers, uneven cell distribution and arrangement in DM4 group, and significantly reduced epidermal cell layers, swollen and blurred cells, increasing cell gap, lack of stratified epidermis arrangement for part of epidermis, atropal and degenerated dermal collagen fiber, significantly decreased subcutaneous fat in DM8 group were observed. The results of Western blotting showed that the protein expression of PGP 9.5 in the plantar skin tissue of DM rats was progressively decreased along with the disease, while the protein level of p-JNK in L5, 6-DRG or S1-DRG showed a gradual increasing trend. PGP 9.5 immunoreactive positive nerve terminal density of plantar skin in DM rats had a negative correlation with the protein level of p-JNK in L5, 6-DRG and S1-DRG (P<0.01), but showed a significant positive correlation with the plantar skin thickness (P<0.01). CONCLUSION: The protein level of p-JNK within L5, 6-DRG or S1-DRG in DM rats shows a progressive enhancement. At the same time, there is a significant change in the skin tissue density and structure. The changes of skin tissue and nerve morphology in DM rat may be related to the activation of JNK/SAPK pathway in L5, 6-DRG or S1-DRG cells. Blocking or inhibiting JNK/SAPK pathway may delay the diabetic peripheral neuropathy and reduce the risk of skin lesions.     

Effect of hydroxylamine on pulmonary arterial hypertension induced by chronic hypoxic hypercapnia in rats

AIM: To explore the effects of hydroxylamine on the pulmonary arterial pressure in chronic hypoxic hypercapnic rats. METHODS: Twenty-four male Sprague-Dawley rats were randomly divided into 3 groups (8 rats in each group): the normal control group (NC), hypoxic hypercapnia+normal saline group (NS), hypoxic hypercapnia+hydroxylamine group (HA). The animals in NS and HA groups were kept in the O₂ (9%-11%) and CO₂ (5%-6%) cabin, 8 h a day and 6 days a week for 4 weeks. Before entering the cabin, the rats in HA group were administered with 1 mL hydroxylamine (12.5 mg/kg) by intraperitoneal injection, while the rats in NS group were given intraperitoneal injection of 1 mL saline solution. The mean pulmonary arterial pressure (mPAP) was measured by external jugular vein cannulation. The heart was removed, and the right ventricle (RV) and the left ventricle plus the septum (LV+S) were dissected. The ratio of the wet weight of the RV to that of the LV+S was calculated. The changes of the pulmonary vascular construction were observed under optical microscope. The concentration of H₂S in the plasma was measured with a spectrometer. The expression of cystathionine-γ-lyase (CSE) in the pulmonary arterioles and bronchi was measured by immunohistochemistry and RT-PCR. RESULTS: The values of mPAP, RV/(LV+S),vessel wall area/total area (WA/TA) and media thickness of pulmonary arterioles (PAMT) in NS group and HA group were significantly higher than those in NC group (P<0.05). The level of H₂S in the plasma, the content of CSE protein and the expression of CSE mRNA in NC group were significantly lower than those in NS group (P<0.05). The values of mPAP, RV/(LV+S), WA/TA and PAMT in HA group were significantly lower than those in NS group (P<0.05). The level of H₂S in the plasma, the content of CSE protein and the expression of CSE mRNA in HA group were significantly higher than those in NS group (P<0.05). CONCLUSION: Hydroxylamine may decrease the pulmonary arterial hypertension induced by chronic hypoxic hypercapnia in rats by increasing the level of H₂S in the plasma, the content of CSE protein and the mRNA expression of CSE, thus improving the pulmonary vascular structural remodeling.     

Effect of diltiazem on pulmonary arterial pressure and ceNOS mRNA expression in pulmonary arteries in chronic hypoxic hypercapnic rats

AIM: To investigate the effect of diltiazem on mean pulmonary arterial pressure (mPAP) and nitric oxide synthase (NOS) in arterioles in chronic hypoxic hypercapnic rats. METHODS: Twenty-four rats were randomly divided into three groups: control group (A), hypoxic hypercapnic group (B), hypoxic hypercapnia+ diltiazem group (C), constitutive endothelial NOS (ceNOS) were observed in arterioles of rats using the technique of immunohistochemistry, ceNOS mRNA were observed by the technique of in situ hybridization. RESULTS: (1) mPAP was significantly higher in rats of B group than that of A and C group(P<0.01). Differences of mCAP were not significant between A group and B groups (P>0.05), but mCAP was lower in rats of C group than that in B group. (2) Light microscopy showed WA/TA (vessel wall area/total area) was significantly lower in rats of C group than that of B group (P<0.01), electron microscopy showed that diltiazem inhibited the proliferation of smooth muscle cells and collageous fibers of pulmonary arterioles in chronic hypoxic hypercapnic rats. (3) Immunohistochemistry showed the average value of integral light density (LD) of ceNOS in pulmonary arterioles was significantly higher in rats of C group than that of B group (P<0.01), in situ hybridization showed LD of ceNOS mRNA in pulmonary arterioles was significantly higher in rats of C group than that of B group (P<0.01). CONCLUSION: Diltiazem inhibited pulmonary hypertension, the proliferation of smooth muscle cells and collagenous fibers of pulmonary arterioles in chronic hypoxic hypercapnic rats by incresing the expression of ceNOS in pulmonary arterioles.     

Relationship between expression of heme oxygenase-1 mRNA and pulmonary hypertention induced by hypoxic hypercapnia

AIM: To study the effect of chronic hypoxic hypercapnia on expression of heme oxygenase-1 (HO-1). METHODS: Sprague-Dawley rats were randomly divided into three groups: control group(A),hypoxic hypercapnic group(B), hypoxic hypercapnia+hemin group(C). HO-1 and HO-1 mRNA were observed in pulmonary arterioles by the technique of immunohistochemistry and in situ hybridization. RESULTS: ① mPAP and weight ratio of right ventricle (RV) to left ventricle plus septum (LV+S) were significantly higher in rats of B group than those of A and C group (P<0.01). Differences of mCAP were not significant in three groups(P>0.05). ② Blood CO concentration was significantly higher in rats of B group than that of A group (P<0.01), it was much higher in C group than that of B group(P<0.01). ③ Light microscopy showed that vessel well area/total area (WA/TA), density of medial smooth muscle cell (SMC) and media thickness of pulmonary arterioles were much higher in rats of B group than those of A and C group (P<0.01). ④ The observation by electron microscopy showed proliferation of medial smooth muscle cells and collageous fibers of pulmonary arterioles in rats of B group, hemin could reverse the changes mentioned above. ⑤ HO-1 and HO-1 mRNA in pulmonary arterioles was significantly higher in rats of B group than those of A group(P<0.01), and they were significantly higher in rats of C group than those of B group (P<0.01). CONCLUSION: Expression of HO-1 mRNA and HO-1 in pulmonary arterioles was enhanced by hypoxic hypercapnia. Hemin partly inhibited pulmonary hypertension and pulmonary vessel remodeling by enhancing the expression of HO-1 mRNA and HO-1.     

Effects of AT1 receptor autoantibody on renal cell apoptosis and JNK expression in diabetic nephropathy rats

AIM:To study the effects of angiotensin Ⅱ type 1 receptor autoantibody (AT1-AA) on the apoptosis of renal cell and the expression of c-Jun N-terminal kinase (JNK) in diabetic nephropathy (DN) rats. METHODS:High-sucrose and high-fat diet and intraperitoneal injection of streptozotocin were utilized to induce DN rat model. We employed enzyme-linked immunosorbent assay (ELISA) for serum AT1-AA and TUNEL staining for renal cell apoptosis. Furthermore, Western blotting was performed to measure the levels of endoplasmic reticulum stress (ERS) chaperone glucose-regulated protein 78 (GRP78) and ERS-associated apoptosis protein p-JNK. RESULTS:The renal cell apoptotic rate in DN group was significantly increased compared with NC group, and the apoptotic renal cells in AT1-AA positive DN rats were much greater than those in AT1-AA negative DN rats (P<0.05). The protein levels of GRP78 and p-JNK were significantly increased compared with NC group. GRP78 and p-JNK protein levels also significantly increased in AT1-AA positive DN rats compared with AT1-AA negative DN rats. CONCLUSION: AT1-AA activates ERS response and induces renal cell apoptosis via the JNK apoptotic pathway in the renal tissues of DN rats.     

Effect of JNK/MCP-1 signaling pathway on anti-diabetic neuropathic pain by curcumin in type 2 diabetic rats

AIM:To investigate the effect of JNK/MCP-1 signaling pathway on anti-diabetic neuropathic pain by curcumin in type 2 diabetic rats. METHODS:The male Sprague-Dawley rats were induced as the model of the type 2 diabetic neuropathic pain rats, they were randomly divided into 6 groups (n=27): type 2 diabetic neuropathic pain (DNP) group, type 2 diabetic neuropathic pain and intraperitoneal injection of curcumin (Cur) group, type 2 diabetic neuropathic pain and solvent control (DSC) group, type 2 diabetic neuropathic pain and JNK inhibitor (DJ) group, type 2 diabetic neuropathic pain and JNK inhibitor solvent control (DJS) group, type 2 diabetic neuropathic pain and monocyte chemoattractant protein 1 (MCP-1) agonist (DM) group. Another 27 normal SD rats were selected as control group. Mechanical withdrawal threshod and thermal withdrawal latency were measured at 3rd d, 7th d and 14th d after dosing, then the lumbar segment 4~6 of the spinal cord and L4~6 DRG were removed at the same time. ELISA was used to measure MCP-1 level. The expression of p-JNK was determined by Western blotting. RESULTS:Compared with DNP group, p-JNK was significantly decreased at 7th d and 14th d in Cur group, DJ group and DM group after treatment (P<0.05). Compared with C group, the MCP-1 was significantly declined in other 6 group after streptozotocin injection (P<0.05). Compared with DNP group, MCP-1 were significantly increased at 7th d and 14th d in Cur group and DJ group after treatment (P<0.05), and that in DM group was greatly decreased (P<0.05). CONCLUSION: The expression of p-JNK and MCP-1 was increased in DNP rats with spinal cord and dorsal root ganglion. The mechanism of curcumin reducing the neuropathic pain in type 2 diabetic rats might be through regulating the JNK/MCP-1 pathway.     

Changes of NO- and H2O2-dependent soluble guanylate cyclase pathway in the hypoxic hypercapnic pulmonary hypertension rats

AIM: To study the effect of hypoxia and hypercapnia on nitric oxide (NO) in plasma and superoxide dismutase (SOD), catalase (CAT), soluble guanylate cyclase (sGC), cyclic guanosine monophospholate (cGMP) in lung tissue in rats, and to explore the effect of NO- and H2O2-sGC pathway on the development of the pulmonary hypertension. METHODS: The model of hypoxic and hypercapnic 1, 2, 4-week group (HH 1 week, HH 2 weeks, HH 4 weeks) and control group was set up. NO content in plasma, CAT and SOD in rat lung were determined by spectrophotometry. The sGC activity in lung tissue was detected by enzyme kinetic analysis. cGMP content in lung tissue was examined with ［125I］-radioimmunoassay. RESULTS: The mean pulmonary artery pressure (mPAP) showed significantly higher in HH 1 week, HH 2 weeks and HH 4 weeks groups compared with control group (all P<0.05). NO concentration in plasma, CAT, SOD, basal or nitroprusside-or H2O2- stimulated sGC activity and cGMP concentration in lung homogenates were significantly lower (P<0.05, P<0.01, P<0.01, respectively) in HH 1 week, HH 2 weeks and HH 4 weeks groups compared with control group. CONCLUSION: The inhibition of NO- and H2O2-sGC pathway by hypoxia and hypercapnia plays an important role in the development of pulmonary hypertension.     

Effect of ligustrazine on hydrogen sulfide system in lung tissue of rats with pulmonary hypertension induced by hypoxic hypercapnia

AIM: To investigate the effect of ligustrazine on hydrogen sulfide (H2S) system in pulmonary hypertension induced by hypoxic hypercapnia in rats. METHODS: Thirty Sprague-Dawley rats were randomly divided into 3 groups: control group (C), hypoxic hypercapnia group (HH), and hypoxic hypercapnia+ligustrazine group (HH+L). The change of hemodynamics was measured. The ratio of vessel wall area and total area of arteriae pulmonalis were observed under light microscope. The apoptosis of arteriae pulmonalis was tested with TdT-mediated dUTP nick end labeling (TUNEL), and the apoptosis index was calculated. Plasma level of hydrogen sulfide and activity of hydrogen sulfide generating enzymes in homogenates of rat lung tissue were evaluated by sensitive modified sulfide electrode method. Cystathionine-γ-lyase (CSE) mRNA in lung tissues was determined by RT-PCR. RESULTS: The level of mean pulmonary arterial pressure, the ratio of vessel wall area/total area and the right ventricle/left ventricle+septum were significantly higher in HH group than those in C group, and the value was obviously lower in HH+LTZ group than that in HH group (all P<0.01). The mean carotid arterial pressure of 3 groups had no significant difference (P>0.05). The apoptotic index of arteriae pulmonalis in HH group and HH+LTZ group was significantly lower than that in C group, and that in HH+LTZ group was significantly higher than that in HH group (P<0.05 or P<0.01). Plasma level of H2S, the activity of H2S generating enzymes in homogenates of rat lung tissue, cystathionine-γ-lyase (CSE) mRNA in lung tissues in HH group were significantly lower than those in C group (all P<0.01), and those in HH+LTZ group were significantly lower than those in HH group (all P<0.01). CONCLUSION: Ligustrazine up-regulates the expression of cystanthionine-γ-splitting enzyme (CSE), enhances the activity of CSE and increases the level of H2S to prevent pulmonary hypertension induced by hypoxic hypercapnia.     

Effect of L-arginine liposome on L-arginine transport of pulmonary artery in rats chronically exposed to hypoxia-hypercapnia

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.     

Effect of ligustrazine on pulmonary arterial pressure and pulmonary arterial wall collagen of chronic hypoxic hypercapnic rats

AIM:To study the effect of ligustrazine on pulmonary hypertensive rats induced by hypoxic hypercapnia. METHODS:Thirty rats were randomly divided into three groups:control group(A),hypoxic hypercapnic group(B), hypoxic hypercapnia+ligustrazine(lig.) group(C). RESULTS: (1) Mean pulmonary arterial pressure(mPAP)of group B was significantly higher than that of group A and mPAP of group C was significantly lower than that of group B(P＜0.01),differences of mean carotid pressure(mCAP) were not significant among three groups (P＞0.05); (2)Electron microscopy and immunohistochemistry showed ligustrazine could inhibit the diposition of collagenous fiber(collagen typeⅠ)in pulmonary arterioles induced by hypoxic hypercapnia; (3) Plasma endothelin level of group C was significantly lower than that of group B (P＜0.01), serum (NO ₂^-/NO₃^-) of group C was significantly higher than that of group B (P＜0.01). CONCLUSION:Ligustrazine can inhibit pulmonary hypertension and the diposition of collagen type Ⅰ in pulmonary arterial wall induced by hypoxic hypercapnia.     

Effect of glucagon-like peptide 1 on Sesn2/AMPK/mTOR signaling pathway in liver of obese rats induced by high-fat diet

AIM: To explored the effect of glucagon-like peptide 1 receptor agonist liraglutide on Sesn2/AMPK/mTOR signaling pathway in the liver of obese rats.METHODS: Male SD rats were divided into normal chow (NC) group (n=12) and high-fat diet (HF) group (n=33). After 12 weeks, 5 rats of each group were used to assess establishment of obese rat model. The rats in HF group were divided into 4 subgroups, HF group, low dose of liraglutide (LG) group, middle dose of liraglutide (MG) group, and high dose of liraglutide (HG) group, and treated with various doses of liraglutide (0, 50, 100 and 200 μg/kg) via hypodermic injection twice a day for 4 weeks. The body weight and epididymal fat index of the rats at the 16th week were measured. The liver tissue fatty degeneration was observed. The protein levels of Sesn2, AMPK, p-AMPK, mTOR and p-mTOR were determined by Western blot.RESULTS: The body weight of rats in HF group was obviously higher than that in NC group (P<0.01). Compared with NC group, the levels of Sesn2 and p-AMPK/AMPK were significantly decreased in HF group (P<0.01), while the level of p-mTOR/mTOR was not changed. After treatment with liraglutide for 4-week, the body weight of the rats in LG, MG and HG groups was obviously lower than that in HF group (P<0.01), and epididymal fat index of the rats in MG and HG groups was obviously lower than that in HF group (P<0.01). The protein level of Sesn2 in HG group was obviously higher than that in HF group (P<0.01). The level of p-AMPK/AMPK was significantly increased in MG and HG groups (P<0.01). The level of p-mTOR/mTOR was significantly increased decreased in LG, MG and HG groups (P<0.01).CONCLUSION: Glucagon-like peptide 1 receptor agonist liraglutide affects energy metabolism and improves the state of obesity through Sesn2/AMPK/mTOR signaling pathway.     

The regulation of nitric-oxide synthase/nitric-oxide system by endogenous carbon monoxide in rats with pulmonary hypertension

AIM:To study the role and the mechanism of heme oxygenas/endogenous carbon monoxide on nitric oxide synthase/nitric oxide system in rats with pulmonary hypertension induced by hypoxic hypercapnia.METHODS:Sprague-Dawley rats were randomly divided into three groups: control group (A group), hypoxic hypercapnic group (B group), hypoxic hypercapnia+hemin group (C group). Blood CO concentration (COHb%), NO concentration, HO-1 activity, iNOS, cNOS in blood serum and lung homogenate were measured, respectively. RESULTS:① mPAP and RV/(LV+S) of B group were significantly higher than those of A and C group(P＜0.01).② Blood CO concentration, activity of HO-1in blood serum and lung homogenate in rats of B group were significantly higher than those of A group, but were significantly lower than those of C group (P＜0.01). ③ NO concentration in blood serum and lung homogenate in rats of B group were significantly lower than those of A group, those of C group were significantly higher than those of B group (P＜0.01).④The activity of iNOS in blood serum and lung homogenate in rats of B group were significantly higher than those of A group, but were significantly lower than those of C group (P＜0.01). Activity of cNOS in blood serum and lung homogenate of B group were significantly lower than those of A group (P＜0.01), and there was no significant difference between cNOS in B and C group.CONCLUSION:Endogenous carbon monoxide upregulated iNOS/NO system in rats with chronic pulmonary hypertension induced by hypoxic hypercapnia.     

All-trans retinoic acid attenuates blood-brain barrier injury induced by cerebral ischemia-reperfusion in rats through JNK/P38 MAPK signaling pathway

AIM: To investigate the effect of all-trans retinoic acid (ATRA) on blood-brain barrier after cerebral ischemia-reperfusion (CIR) injury in rats and its possible role mechanism.METHODS: Male SD rats were randomly divided into sham group, model (CIR) group and CIR+ATRA (10, 30 and 90 mg/kg) groups. The rat model of CIR injury was established by MCAO thread occlusion method. After ischemia for 1.5 h and reperfusion for 24 h, the neurological functional behavioral score, cerebral infarction volume, brain water content and Evans blue content were determined. The activity of matrix metalloprotein-9 (MMP-9) was measured by gelatin zymography. The protein levels of claudin-5, occludin, ZO-1, JNK, p-JNK, P38, p-P38 and MMP-9 in the brain tissues were determined by Western blot.RESULTS: Compared with CIR model group, ATRA at 30 mg/kg significantly improved neurological function, and decreased cerebral infarction volume, brain water content, Evans blue content and the degradation of tight junction proteins in ischemic area (P<0.01). The activity and protein expression of MMP-9 in ischemic brain tissue were decreased (P<0.01). The phosphorylation of JNK and P38 was inhibited and the protein levels of p-JNK and p-P38 were decreased (P<0.01).CONCLUSION: ATRA reduces the damage of brain tissue and the destruction of blood-brain barrier induced by CIR in rats. The protective effect may be related to inhibiting the activation of JNK/P38 MAPK signaling pathway and MMP-9.     

Changes of dendritic morphology and spine density in hippocampal CA1 pyramidal cells of chronic cerebral ischemic rats

AIM: To investigate the changes of dendritic morphology and spine density in hippocampal CA1 pyramidal cells of the chronic cerebral ischemic rats. METHODS: The model of chronic cerebral ischemia was established by permanent occlusion of the bilateral common carotid arteries (2VO) in rats. Two weeks, 4 weeks or 8 weeks later, the behavior of the rats in each group was evaluated through the Morris water maze to select the successful modeling, and the brains were collected for processing Golgi staining. The changes in dendritic branch and length, and spine density in hippocampal CA1 pyramidal cells were observed under optical microscope. RESULTS: Compared with sham-operated group, dendritic branch and length in model group was significantly reduced in 4-week group and 8-week group (P<0.01), and spine density in model group were significantly reduced in 2-week, 4-week and 8-week groups (P<0.01). With prolonged ischemia, dendritic branch and length, and spine density in model group were all significantly reduced (P<0.05). CONCLUSION: Chronic cerebral ischemia leads to traumatic changes in dendrites and spines in hippocampal CA1 pyramidal cells, which constitutes the pathophysiological basis in the progressive cognitive dysfunction.     

Inhibitory effect of notoginsenoside monomer R1 on activation of p38 MAPK signaling pathway induced by hypoxic hypercapnia in PASMCs

AIM: To explore the mechanism of notoginsenoside monomer R₁ (R₁) against hypoxic hypercapnia-induced pulmonary vasoconstriction (HHPV) by investigating the effect of R₁ on p38 mitogen-activated protein kinase (p38MAPK) signaling pathway in pulmonary arterial smooth muscle cells (PASMCs) under the condition of hypoxia and hypercapnia. METHODS: Primary cultured PASMCs, which were isolated from Sprague-Dawley rats, were incubated in logarithmic growth phase from the 2nd to 5th generation with different concentrations (8, 40 and 100 mg/L) of R₁ under the condition of 6% CO₂ plus 1% O₂ for 24 h. The expression of p38 at mRNA and protein levels was detected by RT-PCR and Western blotting,respectively. RESULTS: The results of Western blotting and RT-PCR analysis indicated that the protein and mRNA expression levels of p-p38 MAPK were significantly higher in hypoxic hypercapnia group with DMSO control than those in normoxia control group (P<0.01). In R₁ treatment groups, the levels of p-p38 MAPK protein and p38 MAPK mRNA were markedly decreased (P<0.01) in a dose-dependent manner. CONCLUSION: p38 MAPK signaling pathway may mediate hypoxic hypercapnia pulmonary vasoconstriction in rats. Notoginsenoside monomer R₁ attenuates HHPV, which may be related to blockage of p38 MAPK signal pathway.     

Alterations in rat hippocampal norepinephrine and serotonin levels under physical exercise and psychological stress

AIM: This study is to determine changes of hippocampal norepinephrine (NE) and serotonin (5-HT) in long term physical exercise and chronic psychological stress, and to study the roles of the two monoamine transmitters in the effect of exercise counteracting stress-induced hippocampal damages in brain. METHODS: Levels of hippocampal NE and 5-HT in rats undergoing 4-week voluntary wheel running exercise (exercise group) or 3-week restraint stress (stress group) or 4-week exercise and 3-week stress (exercise-stress group) were detected by high-performance liquid chromatography using electrochemical detection. RESULTS: It is showed that levels of hippocampal NE and 5-HT increased significantly (P<0.01) in the exercised rats, and in the stressed rats,hippocampal 5-HT levels significantly decreased(P<0.05). Additionally, the NE levels maintained significant high (P<0.01) in exercise-stressed rats compared to the pure stressed ones. On the other hand, no obvious difference was observed in hippocampal 5-HT levels between stress group and exercise-stress group, which were all significant lower (P<0.05) than that in exercise group. CONCLUSION: It is suggested that both the NE and 5-HT may play important roles in mediating the exercise-induced positive effects and the 5-HT may play an important role in stress-induced negative effects on the hippocampus. Moreover, NE may take more action in the exercise attenuating stress-induced hippocampal damages. The hippocampal NE may be more susceptible to exercise, and the hippocampal 5-HT may be more susceptible to stress.