Hypertonic NaCl-NaAc improves the microcirculation in rats subjected to hemorrhagic shock

AIM: To study the effects of hypertonic NaCl-NaAc on microcirculation in hemorrhage-shocked rats.METHODS: SD rats were randomized into three groups of 7.5% NaCl(hypertonic saline, HS), 5% NaCl-3.5% NaAc(hypertonic sodium acetate, HSA) and 0.9% NaCl(normal saline, NS). 4 mL/kg HS, HSA or NS was given intravenously following hemorrhagic shock. The microcirculation of spinotrapezius muscle was observed.RESULTS: HS increased mean aortic pressure more significant than HSA. Variables including arteriolar and venular diameter, velocity and volumetric flow rate and open capillaries were increased and erythrocyte aggregation was decreased in 5 min after resuscitation with both HS and HSA solutions.5 min later, variables were deteriorated in HS group.After local treatment, arteriolar and venular diameters were dilated significantly in HSA group.CONCLUSION:HSA had superior effects to HS in improving microcirculation of hemorrhage-shocked rats.     

Changes of lymphatic vessel response to norepinephrine in hemorrhagic shock rats

AIM: To observe the changes of lymphatic vessel response to norepinephrine (NE) in hemorrhagic shock (HS) rats, and to explore the role of lymphatic reactivity in the pathogenesis of shock. METHODS: The lymphatic vessel pressure was observed through intubating into abdomen thoracic duct in 8 rats in sham group and HS group (which was bled from femoral artery until the mean arterial pressure to 40 mmHg). The changes of lymphatic vessel pressure response to NE at different time points were observed by injection of NE (5 μg/kg) through femoral vein. The spontaneous contraction frequency (F), maximal contraction diameter (a), maximal diastolic diameter (b) and static diameter (c) of mesenteric lymphatic (ML) living samples in 8 rats of each group were recorded through microcirculation video systems continuously. The changes of lymphatic fractional contraction index (index I), total contractile activity index (index II) and lymphatic dynamic index (LD-index) (to show the value using △F, △index I, △indexⅡ, △LD-index) were calculated after injection of NE at different time points. RESULTS: The changes of lymphatic boosting pressure response to NE in HS group was started to diminish 30 min after shock, and showed a progressive decreasing trend which significantly reduced than that in sham group at all time points of shock 1 h-3 h. In HS group, the △F, △indexⅡ, △LD-index at shock 1 h, the △F, △index I, △indexⅡ, △LD-index at shock 1.5 h and 2 h were significantly lower than those in sham group, and the △F, △index I, △indexⅡ, △LD-index at all time points were significantly decreased as compared to the values of pre-shock. CONCLUSION: Lymphatic vessel reactivity in shock rats is progressive declined in the process of hemorrhagic shock. The lymphatic vessel hypo-reactivity might play an important role in the pathogenesis of shock.     

CO2 与养分交互作用对番茄幼苗养分利用的影响

以番茄(Lycopersicon esculentumMill.)‘合作906’为材料进行溶液培养试验,设2个因子:CO2和营养液浓度;CO2浓度设正常(360μL/L)和倍增(720μL/L)2个水平;营养液浓度设基本营养液(日本山崎番茄营养液),微量元素采用阿农营养液配方的1/2、1/4、1/8、1/164个水平,完全试验方案8个处理,3次重复。pH为6·0±0·2,3d更换1次营养液。移植到1·2L盆(2株/盒)中,植株在CO2生长箱(VS-3DMC)中培养,全天施放CO2,白天25℃,晚上15℃,光照为14h/d,光照强度11000lx,相对湿度60%。46d时收获,根、茎、叶经蒸馏水冲洗吸干水分后,放入纸袋105℃杀青,75…     

Role of myosin-light-chain kinase in biphasic contractile activity of lymphatics isolated from hemorrhagic shock rats

AIM: To elucidate the mechanism by which myosin-light-chain kinase (MLCK) modulates the biphasic contractile activity of lymphatics isolated from the rats subject to hemorrhagic shock (HS). METHODS: Male Wistar rats were randomiz to control group and HS group. In HS group, the rats were subject to HS and then further divided into HS 0 h, 0.5 h, 1 h, 2 h and 3 h subgroups. Thoracic ducts of control and shock rats were isolated and used to determine the protein levels of phosphorylated MLCK (p-MLCK). In addition, thoracic ducts obtained from control, 0.5 h- and 2 h-shocked rats were used to observe the contractile properties of lymphatics by a pressure myograph in vitro . Lymphatic rings were prepared and incubated with ML-7 (a specific inhibitor of MLCK) or substance P (SP, an agonist of MLCK). During the experiment, the contractile frequency (CF), end-diastolic diameter, end-systolic diameter and passive diameter in Ca²⁺-free PSS buffer were measured and used to calculate the lymphatic tonic index (TI), contractile amplitude (CA) and fractional pump flow (FPF) as the indexes of lymphatic contraction activity. RESULTS: The levels of p-MLCK in lymphatics in 0 h- and 0.5 h-shocked rats were significantly increased compared with the control rats, and it was gradually decreased with the development of shock. The values of CF, TI and FPF in 0.5 h-shocked lymphatics were significantly increased at transmural pressure of 1, 3 and 5 cmH₂O compared with those in control group, and significantly blunted by ML-7. SP obviously increased the suppressive effects induced by ML-7 and restored the values of CF, TI and FPF to the levels of HS 0.5 h group. CF, TI and FPF in 2 h-shocked lymphatics significantly declined under different transmural pressure as compared with those in control group, and significantly elevated by SP. Similarly, ML-7 depressed the effects of SP. No significant difference was found in CA between 0.5 h- and 2 h-shocked lymphatics. SP decreased the CA of lymphatics obtained from 2 h-shocked rats and this effect was suppressed by ML-7. However, both agents had no effects on CA of 0.5 h-shocked lymphatics. CONCLUSION: MLCK, as an essential enzyme that influences the contraction of lymphatic smooth muscle cells, involves in the modulation of biphasic changes of lymphatic contractile activity during the process of HS.     

Mechanism of calcium sensitivity in lymphatic hypo-reactivity in hemorrhagic shock rats

AIM: To observe the changes of lymphatic reactivity to norepinephrine (NE) and calcium sensitivity in vitro in hemorrhagic shock (HS) rats. METHODS: Male Wistar rats were randomly divided into sham group (with only operation), HS group (duplicating HS model, and divided into shock 1 h and shock 2 h subgroups). The thoracic duct rings (n=48 in each group) were prepared for assaying the lymphatic reactivity to NE and calcium sensitivity by lymphatic tension measurement technique in vitro with isolated perfusion system. Meanwhile, the effects of angiotensin Ⅱ (Ang Ⅱ) and insulin (Ins) on lymphatic reactivity were also observed. RESULTS: Compared with sham group, the NE concentration-response curves in HS 1 h and HS 2 h groups, and calcium concentration-response curves in HS 2 h group were obviously shifted to right. The lymphatic reactivity to NE, contraction to calcium, maximum effect(E_max)and avidity index (pD₂) were markedly reduced. In HS group, after incubating with calcium sensitizer Ang Ⅱ, the lymphatic reactivity to NE and calcium sensitivity were significantly increased but reduced in sham group. However, calcium sensitivity inhibitor Ins decreased the lymphatic contractile response to NE and Ca²⁺. CONCLUSION: The lymphatic hypo-reactivity in hemorrhagic shock rats is related to calcium desensitization, indicating a mechanism of lymphatic hypo-contraction.     

Substance P enhances pump function of isolated lymphatics from hemorrhagic shock rats

AIM: To establish a technique of lymphatic perfusion in vitro and to determine the effect of substance P (SP) on lymphatic contractility during the process of hemorrhagic shock (HS). METHODS: Male Wistar rats were randomly divided into control group (surgical procedure only) and HS group . Thoracic ducts were isolated from HS rats at the corresponding time points. The segment of thoracic duct was prepared, perfused in vitro at the transmural pressure of 3 cmH₂O and stimulated with gradient concentrations of SP to measure the lymphatic contractile activity. The end-systolic diameter, end-diastolic diameter, contraction frequency (CF) and passive diameter of isolated lymphatics were measured. The contraction amplitude (CA), tonic index (TI) and fractional pump flow (FPF) were also calculated.RESULTS: SP increased lymphatic CF, TI and FPF, and the effects were enhanced with the increase in the concentration of SP. The CF, TI and FPF of 2 h- and 3 h- shocked lymphatics were elevated to/over the value of baseline levels before the experiment by SP at the concentration starting from 3×10^-8 mol/L. At the same concentration of SP, the CA of lymphatics showed no significant statistical difference among the groups. However, with the increase in SP concentration, the lymphatic CA had a downward trend in all groups.CONCLUSION: SP enhances the pump function of lymphatics not only under physiological condition, but also in shock during different stages.     

Dexmedetomidine attenuates acute kidney injury induced by hemorrhagic shock/resuscitation in rats

AIM: To investigate the effects of dexmedetomidine on hemorrhagic shock/resuscitation (HS/R)-induced acute kidney injury (AKI) in rats, and to explore the possible mechanisms. METHODS: Wistar rats (n=32) were randomly divided into 4 groups (n=8):normal saline control group (NS group), dexmedetomidine group (D group), HS/R group and HS/R+D group. The animals were sacrificed at 6 h after resuscitation. The levels of serum creatinine (Cr) and blood urine nitrogen (BUN) were examined. The kidneys of all rats were removed for evaluation of histological characteristics, and the levels of malondialdehyde (MDA), tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) and superoxide dismutase (SOD) were measured. The expression of nuclear factor-κB (NF-κB) and hemeoxygenase-1 (HO-1) was determined by Western blot. RESULTS: Compared with NS group, the levels of Cr, BUN, MDA, TNF-α and IL-1β were obviously increased in HS/R group, which were obviously decreased in HS/R+D group (P<0.05). Compared with NS group, the SOD activity was obviously decreased in HS/R group, which was obviously increased in HS/R+D group (P<0.05). Compared with NS group, the protein expression of NF-κB was obviously increased in HS/R group, which was obviously decreased in HS/R+D group (P<0.05). Compared with NS group, the protein expression of HO-1 was increased in HS/R group. Compared with HS/R group, the protein expression of HO-1 was obviously increased in HS/R+D group. Compared with NS group, HS/R induced marked kidney histological injury, which was less pronounced in HS/R+D group.CONCLUSION: Dexmedetomidine effectively protects rats against AKI caused by HS/R, and its mechanism may be associated with the increase in HO-1 expression and the inhibition of NF-κB expression.     

Role of nitric oxide in reactivity of isolated lymphatics to substance P in shock rats

AIM: To observe the role of nitric oxide (NO) in the reactivity of isolated lymphatics to substance P (SP),which presents a biphasic change, in the hemorrhagic shock (HS) rats with the technique of lymphatic perfusion in vitro. METHODS: Male Wistar rats were randomly divided into control group (surgical procedure only) and shock group (the rats were further divided into shock 0.5 h and shock 2 h groups after the HS model was established). A segment of lymphatics was pressed and perfused in vitro at transmural pressure of 3 cmH₂O after thoracic ducts were separated from the rats at the corresponding time points in each group. The lymphatics of shock 0.5 h and shock 2 h were incubated with different drugs for changing the activity of No and nitric oxide synthase (NOS), respectively. The end-systolic diameter, end-diastolic diameter, contraction frequency (CF) and passive diameter of isolated lymphatics were measured, while the contraction amplitude (CA), tonic index (TI) and fractional pump flow (FPF) were calculated after stimulated with gradient SP. Different values between pre-and post-administration of SP in CF, CA, TI and FPF were calculated and expressed as ΔCF, ΔTI, ΔCA and ΔFPF for further assessing the reactivity of lymphatics. RESULTS: NO donor L-Arg reduced ΔCF, ΔTI and ΔFPF of 0.5 h-shocked lymphatics treated with different concentrations of SP. The effect of L-Arg was obviously suppressed by a soluble guanylate cyclase inhibitor ODQ. ΔCF, ΔTI and ΔFPF increased strikingly compared with shock 0.5 h+L-Arg group in the presence of SP at certain concentration, and ΔCF and ΔFPF increased remarkably compared with control group. NOS inhibitor L-NAME elevated ΔCF, ΔTI and ΔFPF of 2 h-shocked lymphatics treated with different concentrations of SP and the manifestation of lymphatics exceeded the values of control levels. In the experiment of 2 h-shocked lymphatics treated with L-NAME+phosphodiesterase inhibitor aminophylline (AP), the effect of L-NAME was suppressed significantly, which manifested by the decrease in ΔCF, ΔTI and ΔFPF as compared with the values of shock 2 h+L-NAME group in the presence of SP at the concentrations of 1×10^-8 mol/L and 3×10^-8 mol/L. CONCLUSION: These data indicate that NO involves in the biphasic modulation of shocked lymphatics and the effect might be involved in the action of cyclic guanosine monophosphate.     

Potassium antagonizes damage of coronary artery induced by high salt intake

AIM：To investigate the effect of potassium treatment on coronary arterial impairment induced by high salt intake. METHODS：Sprague-Dawley rats (4-week-old, n=10 in each group) received distilled water (NS), water containing 1.5% NaCl (HS), or 1.5% NaCl and 0.5% KCl (HS+HP) for 16 weeks. Systolic blood pressure (SBP) was determined by tail plethysmography every 2 weeks. After 16 weeks of treatment, vascular remodeling, superoxide production, malondialdehyde (MDA) content, and endothelial nitric oxide synthase (eNOS) and gp91 expression in the coronary arteries were detected. RESULTS：After 16 weeks of salt loading, the rats in HS group was divided into salt sensitive subgroup and salt resistance subgroup according to the tail-cuff blood pressure. In this experiment, the salt-sensitive rats were selected as HS group. In HS group, salt loading significantly increased SBP, serum MDA and gp91 expression, decreased serum NO and eNOS expression in the coronary arteries, and induced the coronary artery remodeling compared with NS group. In salt-loaded SD rats, 16-week potassium treatment abrogated the effects induced by salt loading. CONCLUSION：High salt may affect structural and functional changes in coronary arteries by activating oxidative stress. Potassium treatment antagonizes the effect of high salt intake.     

Effects of hypotensive or aggressive resuscitation on tissue ATPase activity in pregnant rabbits with uncontrolled hemorrhagic shock

AIM: To compare the effects of hypotensive and aggressive resuscitation strategies on blood loss, fluid requirements, hematocrit (Hct), tissue Na＋-K＋-ATPase and Ca2＋-Mg2+-ATPase activities in a clinically relevant model of uncontrolled hemorrhagic shock in pregnancy rabbits. METHODS: Thirty anesthetized New Zealand white rabbits at mid and late gestation underwent uncontrolled hemorrhagic shock by transecting a small artery of mesometrium, followed by bleeding via carotid artery to mean arterial pressure (MAP) of 40-45 mmHg. Animals were randomly divided into five groups (n=6 each): sham shock (SS); shock without resuscitation (SH); aggressive resuscitation in pre-hospital phase with 4 mL/kg normal saline, followed by Ringer’s solution to MAP of 80 mmHg (NS), hypotensive resuscitation with 4 mL/kg of normal saline (NH) or hypertonic hydroxyl ethyl starch (7.5% NaCl + hydroxy ethyl starch, HHES, HHH) followed by Ringer’s solution to MAP of 60 mmHg. Finally, all the resuscitated animals received hemorrhage controlled and fully resuscitated to MAP of 80 mmHg. At the end of the experiment, survivors were sacrificed, skeletal muscle, cardiac muscle, liver, kidney, lung and ileum were harvested for determination of Na＋-K＋-ATPase and Ca2＋-Mg2+-ATPase activities. RESULTS: Total blood loss and infused volume were compared between NH［(4.3±0.2)mL/kg, (47.2±4.1)mL/kg］ and HHH［(4.1±0.3)mL/kg,(44.9±4.3)mL/kg］ groups, both were significantly less than NS［(5.5±0.2)mL/kg, (65.5±3.8）mL/kg］　group. Hct in NH (21.0%±2.1%) and HHH (21.5%±1.8%) were significantly higher than NS (14.2%±1.5%) and SH (12.5%±1.4%).Tissue Na＋-K＋-ATPase and Ca2＋-Mg2+-ATPase activities were stimulated in all shock groups. Na＋-K＋-ATPase in skeletal muscle, cardiac muscle, liver, kidney was significantly lower in the NH (5.42±1.41, 4.54±2.01, 4.13±0.62, 3.42±0.84) and HHH (3.97±0.91, 2.94±0.66, 3.22±1.42, 3.03±0.53) than that in NS (7.34±1.41, 6.23±1.53, 6.11±0.97, 5.82±0.69) and SH (9.11±0.52, 8.40±1.08, 7.04±1.13, 6.55±1.45). CONCLUSION: Hypotensive resuscitation with normal saline or HHES reduces blood loss, decreses total infused volume, leads to higher hematocrit and finally alleviates metabolism derangement after uncontrolled hemorrhagic shock.     

Experimental study on pathogenesis of salt-induced hypertension

AIM:To reveal the pathogenesis of salt-induced hypertension.METHODS:Forty male SD rats were divided into five groups, which received on same chow but different drink. Control(NC)group: deionized water; High salt(HS)group: 1.5% NaCl solution; L-arginine(HS+Arg)group: L-arginine(4 g·kg^-1·d^-1)in 1.5% NaCl solution; Enalapril (HS+En) group: enalapril (30 mg·kg^-1·d^-1) in 1.5% NaCl solution; Terazozin(HS+Ter)group: terazozin(4 mg·kg^-1·d^-1)in 1.5% NaCl solution. At the end of 8 weeks, rats were anesthetized with pentobarbital sodium. Blood pressure(BP)were recorded and blood were drawn from inferior vena cava and kidneys, adrenals were removed. NO(x),ET and AngII, Na^＋-K^＋-ATPase and proscillaridin-like compound(PLC)were assayed.RESULTS:BP, PLC and ET in plasma and AngII in adrenal were increased, NO(x)and AngII in plasma and kidney were decreased in HS group compared with NC group.CONCLUSION:High salt intake may induce hypertension in SD rats. In addition to the Na^＋-K^＋-ATPase activity was inhibited by increased sodium-pump inhibitors, NO release decrease may also play an important role in the pathogenesis of hypertension.     

Blockage of mesenteric lymph return promotes the vascular reactivity and calcium sensitivity in hemorrhagic shock rats

AIM: To observe the effects of mesenteric lymph duct ligation and mesenteric lymph drainage on the vascular reactivity and calcium sensitivity in hemorrhagic shock (HS) rats, and to investigate the role of mesenteric lymph on the vascular hyporeactivity during shock. METHODS: Seventy-two male Wistar rats were randomly divided into sham group (only operation), shock (duplicating HS model) group, shock+ligation group (duplicating HS model and mesenteric lymph duct ligation) and shock+drainage group (duplicating HS model and mesenteric lymph drainage). The changes of mean artery pressure (MAP) after injection of norepinephrine (NE, 3 μg/kg) at different time points were recorded. After hypotension (40 mmHg) for 3 h, the vascular ring of superior mesenteric artery (SMA) was made for determining the vascular reactivity and sensitivity to calcium by observing the contraction initiated by NE and Ca²⁺ under depolarizing conditions (120 mmol/L K⁺) in the isolated organ perfusion system. Meanwhile, the effects of angiotensin Ⅱ (AngⅡ) and insulin (Ins) on the vascular reactivity were also observed. RESULTS: Compared to sham group, the △MAP in shock group was increased significantly at 0 h and 0.5 h after shock, and that was decreased markedly at 1.5 h, 2 h, 2.5 h and 3 h after shock, respectively, and that in shock+ligation group and shock+drainage group was increased at 0 h, 0.5 h and 1 h after shock, decreased at 2.5 h and 3 h after shock, respectively. The △MAP in shock+ligation group and shock+drainage group was higher than that in shock group at 0.5 h after shock and all the time points followed. The SMA reactivity to NE and sensibility to Ca²⁺ in shock group, shock+ligation group and shock+drainage group were lower markedly than those in sham group. The vascular reactivity and calcium sensitivity in shock+ligation and shock+drainage groups were higher than those in shock group. The vascular reactivity and calcium sensitivity in shock group, shock+ligation group and shock+drainage group were lower than those in sham group, and those in shock+ligation and shock+drainage groups were increased as compared to shock group, respectively. CONCLUSION: Blockage of mesenteric lymphatic return with the methods of mesenteric lymph duct ligation and mesenteric lymph drainage promotes the vascular reactivity of HS rats. The mechanism may be related to improving the calcium sensitivity in the vasculature.     

Estrogen treatment enhances mesenteric lymphatic contractility in rats after hemorrhagic shock

AIM To investigate the effects of 17β-estradiol （E2） treatment on the mesenteric lymphatic microcirculation and isolated lymphatic contractility in rats after hemorrhagic shock, and to explore the relationship between contractility and the difference between intra- and extracellular calcium ion concentrations （［Ca²⁺］） of lymphatic smooth muscle cells （LSMCs）. METHODS Male Wistar rats were divided into sham group, shock group and shock+E2 group. The rats were subjected to hemorrhage ［（40±2） mmHg for 90 min］ and resuscitation with or without subcutaneous injection of E2 （2 mg/kg）. After resuscitation for 3 h, the mesenteric lymphatic microcirculation in vivo was observed. Moreover, the isolated mesenteric microlymphatic rings were prepared for the observations of lymphatic contractility evaluated by the indexes including end-systolic diameter, end-diastolic diameter, contraction frequency （CF） and passive diameter. Meanwhile, the difference between intra- and extracellular ［Ca²⁺］ of LSMCs was recorded during lymphatic contraction. RESULTS Treatment with E2 significantly enhanced the CF, total contractile fraction and lymphatic dynamics index in vivo in the rats after hemorrhagic shock, and increased the CF, the fractional pump flow and the difference between intra- and extracellular ［Ca²⁺］ of LSMCs in isolated lymphatics from the shocked rats （P<0.05）. CONCLUSION Estrogen treatment enhances lymphatic contractility in rats after hemorrhagic shock, which is related to enhancement of difference between intra- and extracellular ［Ca²⁺］ of LSMCs.     

Effect of caveolin-1 expression on high-salt diet-induced endothelial dysfunction in type 1 diabetic rats

AIM: To investigate the underlying mechanisms responsible for endothelial dysfunction of type 1 diabetes mellitus (DM) rats fed with high-salt diet. METHODS: Type 1 DM was induced by intraperitoneal injection of streptozotocin (70 mg/kg). Normal and diabetic rats were fed high-salt food (HS, 8% NaCl) and standard food for 6 weeks, respectively. Isometric tension of the mesenteric arteries were measured. The expression of Akt, endothelial nitric oxide synthase (eNOS) and caveolin-1 (Cav-1) was examined by Western blot. RESULTS: The rats in DM+HS group exhibited more pronounced impairment of vasorelaxation to acetylcholine and insulin compared with either DM group or HS group (P<0.01). Akt and eNOS phosphorylation levels, and nitric oxide (NO) concentration in DM+HS group were significantly lower than those in DM group (P<0.01). The level of Cav-1 in DM+HS group was significantly higher than that in DM group and HS group. CONCLUSION: Impaired endothelial Akt activation, increased Cav-1 expression and resultant decreased eNOS activation contribute to aggravate high-salt diet-induced endothelial dysfunction and hypertension in DM rats.     

Pinacidil pretreatment increases vascular reactivity and calcium sensitivity after hemorrhagic shock

AIM: To observe the protective effects of protein kinase Cα(PKCα) and protein kinase Cε(PKCε) activated by pinacidil pretreatment on vascular reactivity and calcium sensitivity after hemorrhagic shock in rats. METHODS: The changes of the pressor effect(the change of mean arterial pressure) and vasoconstriction response(the changes of diameter) of superior mesenteric artery(SMA) to norepinephrine(NE) were observed. The vascular reactivity and calcium sensitivity of the first class arborization of SMA induced by pinacidil pretreatment with different volume and at different time points before shock were determined. The effects of PKCα and PKCε antagonists on the protection of pinacidil pretreatment, and the effects of pinacidil pretreatment on the translocation of PKCα and PKCε were also measured. RESULTS: (1) The pressor effect and vasoconstriction response of SMA to NE, and the vascular reactivity and calcium sensitivity of the first class arborization of SMA in 2 h shock group were significantly decreased as compared to those in normal controls(P<0.01). Pinacidil(25 μg/kg) pretreated at 30 min before shock attenuated the above changes.(2) The inhibitors of PKCα and PKCε suppressed the protective effects of pinacidil pretreatment(25 μg/kg pinacidil pretreated at 30 min before shock) on the vascular reactivity and calcium sensitivity. The E_max of NE was decreased by 42.9% and 62.9%, respectively(P<0.01). The E_max of Ca²⁺ was decreased by 31.1% and 56.1%, respectively(P<0.01). Pinacidil(25 μg/kg) pretreated at 30 min before shock increased the protein expression of PKCα and PKCε on the membrane, and decreased the protein expression in the cytoplasm as compared to those in 2 h shock group(P<0.01). CONCLUSION: Pinacidil pretreatment activates PKCα and PKCε, and induces the increasing effects of vascular reactivity and calcium sensitivity after hemorrhagic shock in rats.     

Protective effects and mechanism of heat shock response on cardiovascular system in rats after heat exposure

AIM:To study protective effects and mechanism of heat shock response (HSR) on cardiovascular system in rats after heat exposure.METHODS:The study was divided into 2 experiments:①Protective effects of HSR on cardiovascular system in rats after heat exposure.SD rats randomly allocated into 2 groups:heat shock group (HS group), sham control group(SC group).HS group were treated with heat shock, but SC group weren't.After re-covering for 20 h at room temperature, two groups exposed to death in thermal environment, and blood pressure and elec-trocardiogram were measured continuously.Through Chart software mean arterial pressure(MAP), existent time etc were acquired.②SD male rats randomly allocated into 3 groups:HS group, SC group and normal temperature control group(NC group).NC group weren't treated.The treatment in HS and SC group was identical with in the first experiment, but it would be terminated at 73 min after heat exposure, meanwhile content of MDA of myocardium were measured.RESULTS:① Existent time in HS group was longer than that in SC group and shock arrived later; ② During earlier period after heat exposure MAP had no significant changes between HS and SC group, but after 60 mins MAP in HS group were higher than that in SC group; ③ Compared with NC group, content of MDA in myocardium in SC group was higher significantly at 73 min after heat exposure. Howerer, content of MDA in HS group was lower than in SC group, and had no significant changes with NC group.CONCLUSION:Through decreasing production of MDA in myocardium, HSR has a protective effect on cardiovascular system in rats after heat exposure.     

Role of ZIPK in the regulatory effects of PKCα and PKCε on calcium sensitivity during hemorrhagic shock in rats

AIM: To observe the role of zipper-interacting protein kinase (ZIPK) in the regulatory effects of protein kinase Cα (PKCα) and protein kinase Cε (PKCε) on calcium sensitivity during hemorrhagic shock(HS) in rats. METHODS: The skinned first class arborization of superior mesenteric artery (SMA) from HS rats were adopted to observe the influence of inhibitor of ZIPK on the effects of PKCα and PKCε agonists on calcium sensitivity after shock via measuring the contraction initiated by Ca2+ with isolated organ perfusion system, hypoxic vascualr smooth muscle cells (VSMCs) were adopted to measure the protein expression and activity of ZIPK after applying PKCα and PKCε agonists following hypoxia via Western blotting. RESULTS: (1) The calcium sensitivity of SMA was decreased after 2 h shock, and increased by agonists of PKCα and PKCε. Emax of Ca2+ was increased from 47.2％to 66.5％ (P<0.01) and 66.3％ (P<0.01) of normal control respectively as compared with 2 h shock group. The increasing effects of PKCα and PKCε agonists on calcium sensitivity of SMA after 2 h shock were weakened by the inhibitor of ZIPK. The cumulative dose-response curve of Ca2+ was shifted to the right, the Emax of Ca2+ was decreased to 42.6％ and 47.5％ of normal control (P<0.01), respectively. (2) The protein expression and activity of ZIPK in VSMCs were decreased after 2 h hypoxia, and were increased by the agonists of PKCα and PKCε following 2 h hypoxia. CONCLUSION: PKCα and PKCε regulate the calcium sensitization probably through changing the protein expression and activity of ZIPK following HS in rats.     

Effects of hypoxia exposure for different time on structure and function of red blood cells in rats

AIM: To investigate the effects of hypoxia exposure on the structure and function of erythrocytes in rats at different time. METHODS: Male SD rats (n=40) were randomly divided into 5 groups, normal control group, 1-week hypoxia group, 2-week hypoxia group, 3-week hypoxia group and 4-week hypoxia group, with 8 rats per group. The rats in hypoxia groups were placed in the simulated 5 800 m of high altitude in a hypobaric chamber for different time. The values of detected blood, erythrocyte deformation index, erythrocyte osmotic fragility, erythrocyte oxygen dissociation, erythrocyte apoptosis and bone marrow biopsy were determined. RESULTS: Compared with normal control group, the red blood cell count, hemoglobin content, mean corpuscular volume and mean corpuscular hemoglobin significantly increased (P<0.01). Eversion rate of phosphatidylserine of erythrocytes increased. Oxygen half-saturation of hemoglobin increased (P<0.05). Bone marrow erythroid proliferation increased. The erythrocyte deformation index and erythrocyte osmotic fragility decreased significantly (P<0.01). In addition, oxygen dissociation curves shifted to the right. CONCLUSION: In the early stage of hypoxia, compared with normal control group, the changes of erythrocyte structure and function increase the oxygen supply to the tissue and are conducive to adapting to the plateau. However, with the extension of hypoxia, excessive erythrocytosis results in thrombosis, microcirculation disturbance and aggravating tissue hypoxia.     

Mechanism of vascular hyporeactivity induced by NO in hemorrhagic shock

AIM: To investigate the role of nitric oxide (NO) in vascular hyporeactivity during prolonged hemorrhagic shock (HS). METHODS: Anesthetized Sprague-Dawley rats (180-220 g) were subjected to HS insult in which they were bled to a mean arterial pressure (MAP) of 40 mmHg (5.33 kPa) and arteriolar reactivity to norepinephrine in spinotrapezius was detected. The constant MAP of 40 mmHg was maintained until vascular hyporeactivity had occurred and then were resuscitated or sacrificed for further analysis. NO synthase (NOS) activity was measured ex vivo by the conversion of [³H]-arginine to [³H]-citrulline in homogenates from heart, lung, liver, spleen, duodunum, skeletal muscle. 24 h survival rates of resuscitated rats were observed with and without administration of aminoguanidine (AG), a selective inducible NOS (iNOS) inhibitor. Mesenteric arteriolar smooth muscle cells (ASMC) were isolated, and the effects of L-arginine (L-Arg) on membrane potential (MP) of ASMC were determined by fluorescent probe and confocal microscopy in the absence and presence of AG. RESULTS: When vascular hyporeactivity occurred, an increase of NOS activity was observed in liver and heart. Resuscitated rats with AG had a higher survival rate compared with that of control. The MP of ASMC was decreased (more negative) immediately following the addition of L-Arg, and the hyperpolarization effects of L-Arg were partially blocked in the presence of AG. CONCLUSION: These results suggest that excessive NO produced in HS is responsible for the occurrence of vascular hyporeactivity in prolonged hemorrhagic shock, and one of the mechanisms of which may be hyperpolarization of ASMC caused by NO.     

Mesenteric lymph drainage alleviates spleen injury in hemorrhagic shock rats

AIM：To observe the effects of post-shock mesenteric lymph (PSML) drainage on histopathology, apoptosis, cell cycle and proliferation of the spleen in rats with hemorrhagic shock. METHODS：Eighteen Wistar rats were randomly divided into sham, shock and shock+drainage groups (n=6 in each group). The hemorrhagic shock model was established in the shock and shock+drainage groups. Fluid resuscitation for 30 min was performed 1.5 h after hypotension, and PSML was drained in the rats in shock+drainage group from 1 h after hypotension to 3 h after resuscitation finished. The fixed spleen tissue was harvested from each rat for histological observation with HE staining. The apoptosis of splenocytes was observed by Hoechst 33258 staining. The expression of Bcl-2 and Bax proteins was detected by immunohistochemical staining. The cell cycle and the expression of p53 protein were measured by flow cytometry, and the proliferation index (PI) was calculated. RESULTS：Compared with sham group, splenic tissue injury appeared in the shocked rats. The apoptotic cells and the expression of Bax and p53 in shock group were increased, while Bcl-2 expression was decreased. The percentage of G2/M cells in shock group was decreased. Compared with shock group, the splenic tissue damage in shock+drainage group was significantly attenuated. Moreover, the number of apoptotic cells, the percentage of G0/G1 cells, and the expression of Bax and p53 were obviously decreased, and the G2/M cells, Bcl-2 protein expression and PI were significantly increased in shock+drainage group. CONCLUSION: PSML drainage alleviates splenic injury in hemorrhagic shock rats, which may be related to reducing the apoptosis of splenocytes.