Protective effects of nerve growth factor vs Danshen on hippocampal neuron against global ischemia-reperfusion injury in gerbils / 南方医科大学学报

<p><b>OBJECTIVE</b>To study the protective effects of nerve growth factor (NGF) and Danshen on hippocampal neurons in gerbils (Meriones unguiculatus) with global ischemia-reperfusion injury.</p><p><b>METHODS</b>Global ischemia-reperfusion model was established in 54 male Z:ZCLA gerbils by occlusion of the bilateral carotid arteries. The animal models were randomized into 3 groups to receive treatment with normal saline, NGF, and Danshen 30 min after the reperfusion. At 6 h, 3 and 7 days after the reperfusion, the survival of the hippocampal CA1 pyramidal neurons was observed using optical and electron microscopy, and immunohistochemistry was employed to detect the expressions of Bcl-2 and Bax in the neurons.</p><p><b>RESULTS</b>Neuronal apoptosis was not observed in the hippocampus 6 h after the reperfusion, but at 3 and 7 days, the number of apoptotic neurons increased significantly in the CA1 region. Compared with normal saline, treatments with NGF and Danshen both significantly reduced the number of apoptotic neurons at 3 and 7 days. The number of apoptotic neurons showed no significant difference between NGF and Danshen treatment groups at 3 days, but at 7 days, the apoptotic cell number was significantly lower in NGF group (P<0.05). Bcl-2 expression was the highest in NGF group, and its highest expression occurred at 6 h after the reperfusion; Bax expression was detected in saline group, and underwent no significant changes with the passage of time.</p><p><b>CONCLUSION</b>Both NGF and Danshen show protective effects against global ischemia-reperfusion injury. NGF has a stronger protective effect than Danshen, and this finding provides experimental evidence for selecting appropriate protective agents in the treatment of ischemic brain damage.</p>

The mechanism of signal extension in Haoqin Qingdan decoction immunity activity in Damp-heat syndrome of pneumonia disease infected by influenza virus / 中华预防医学杂志

<p><b>OBJECTIVE</b>To explore the immunoregulation existing signal transduction mechanism, to evaluate the role of lay its experimental basis By using Haoqin Qingdan decoction for treatments on the mouse models.</p><p><b>METHODS</b>A total of 40 NIH Mice were randomly divided into five groups: control group, virus group (infecting by influenza virus), complex model group (richly fatty and sweet diet + Humid heat environment + infecting by influenza virus), virazole group (mouse of model group was treated by virazole), and Haoqin Qingdan decoction group (mouse of complex model group was treated by decoction of Haoqin Qingdan). When the complex model was established, determination of the mice lung indexes in each group and calculate the inhibition of lung indexes. The level of TLR2 mRNA and NF-κB mRNA expressions of peritoneal macrophages in each group of mice were quantitated by reverse transcription-polymerase chain reaction (RT-PCR). The level of IL-4 and IFN-γ in mouse serum was detected by ELISA to calculate the Th1/Th2 (IFN-γ/IL-4).</p><p><b>RESULTS</b>The lung index of control group, virus group, complex model group, virazole group and Haoqin Qingdan decoction group were separately: (0.79 ± 0.11)%, (1.93 ± 0.38)%, (1.41 ± 0.26)%, (1.10 ± 0.26)% and (1.02 ± 0.16)%; The mice of virazole group and Haoqin Qingdan decoction group lung index were decreased (t = 0.322, P < 0.05). TLR2 mRNA expression The results showed that the control group, virus group, complex model group, virazole group and Haoqin Qingdan decoction group were: 0.145 ± 0.017, 0.991 ± 0.149, 0.903 ± 0.124, 0.257 ± 0.03 and 0.413 ± 0.031; Compared to the complex model group, Haoqin Qingdan decoction group and virazole group were decreased (t = 0.422, F = 112.834, P < 0.05). Control group, virus group, complex model group, virazole group and Haoqin Qingdan decoction group NF-κB mRNA expression were separately: 0.075 ± 0.148, 0.379 ± 0.019, 0.291 ± 0.012, 0.169 ± 0.026 and 0.175 ± 0.033; the expression in virazole group and Haoqin Qingdan decoction group were decreased (t = 0.422, F = 112.834, P < 0.05). The level of IFN-γ in mice serum of control group, virus group, complex model group, virazole group and Haoqin Qingdan decoction group were: (7434.06 ± 323.27) pg/ml, (8679.77 ± 198.70) pg/ml, (8068.78 ± 113.8) pg/ml, (7454.66 ± 301.30) pg/ml and (7484.56 ± 229.85) pg/ml respectively; the IFN-γ level in serum of Haoqin Qingdan decoction group and virazole group were decreased (t = 0.201, F = 5.390, P < 0.05). Each group of mice IL-4 contents were (3701.74 ± 256.00) pg/ml, (3569.64 ± 161.35) pg/ml, (3530.88 ± 334.63) pg/ml, (3481.84 ± 282.25) pg/ml and (3618.00 ± 262.16) pg/ml; there were no significant difference between each group (t = 0.414, F = 0.505, P > 0.05). Th1/Th2 type cells in state of equilibrium (means IFN-γ/IL-4) were: 2.02 ± 0.19, 2.38 ± 0.10, 2.36 ± 0.14, 2.22 ± 0.17 and 2.07 ± 0.15; and complex model group Haoqin Qingdan decoction group and virazole group were decreased, and there was no significant difference observed (t = 0.587, F = 3.684, P > 0.05).</p><p><b>CONCLUSION</b>The effect of Haoqin Qingdan decoction on treatment of damp-heat syndrome of pneumonia infected by influenza virus was observed. Through reducing the expressions of TLR2, it decreases the levels of NF-κB mRNA and the proportionality of Th1/Th2 are obviously descend (P < 0.05). Haoqin Qingdan decoction can reduce the lung index and relieve the pathogenic changes.</p>

Construction of MHV-A59 damp-heat mouse model and analysis of the relevant indices / 南方医科大学学报

<p><b>OBJECTIVE</b>To explore the impact of inflammation, water metabolism and immune function on the establishment of a mouse model of damp-heat syndrome with MHV-A59 infection.</p><p><b>METHODS</b>Twenty-four mice were randomly divided into control group, virus group, damp-heat group and model group. The peripheral blood CD4(+) and CD8(+) lymphocytes were detected by flow cytometry, and the serum levels of IFN-γ and IL-4 were assayed by ELISA. The expressions of NF-κB and AQP4 in the liver and stomach were determined using immunohistochemistry.</p><p><b>RESULTS</b>The expression of NF-κB and CD4(+)/CD8(+) ratio in the virus and model groups were significantly higher than those in the damp-heat and control groups, while the expression of AQP4 was significantly higher in the model and damp-heat groups than in the other groups. Compared with the control group, the model group showed a significantly higher ratio of IFN-γ/IL-4.</p><p><b>CONCLUSIONS</b>MHV-A59 virus is the main cause of elevated NF-κB expression and CD4(+)/CD8(+)/ ratio, while damp-heat syndrome is responsible for increased AQP4 expression, and their synergistic effect results in increased IFN-γ/IL-4 ratio. The mouse model established using MHV-A59 virus and the damp-heat factors can mimic damp-heat syndrome described in traditional Chinese medicine theory.</p>

Effects of selective iNOS inhibitor aminoguanidine on waveform of blood pressure in rat heat stroke / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To evaluate the change of blood pressure, ECG and nitric oxide (NO) in rat heat stroke and effects of aminoguanidine (AG) against heatstroke.</p><p><b>METHODS</b>The male SD rats were randomly assigned into 1 of the following 2 groups: control group or AG group. The rats of control group (n = 10) and AG group (n = 10) were exposed to high ambient temperature (41 degrees C, relative humidity 65%) to induce heatstroke, arterial blood pressures, colonic temperature (T(co)), electrocardiograph (ECG) were monitored. The other rats of both groups (both n = 10) were exposed to high ambient temperature (41 degrees C, relative humidity 65%), and the blood samples were taken at 0, 60 min after the start of heat exposure for determination of the plasma NO concentrations.</p><p><b>RESULTS</b>(1) From 0 min to 50 min after heat exposure, MAPs of two groups were not significantly different, but at about 55 approximately 60 min after the start of heat exposure, MAPs of control group were decreased significantly differently from that of AG group, K value and dicrotic pulse relative height (h(D)/H) were gradually decreased, especially at 40 min after the start of heat exposure, K value of control group decreased significantly comparison with that of AG group; (2) Heart rate (HR) and QT interval of both groups were increased, while PR interval were decreased after the start of heat exposure; (3) T(co) of both groups were increased after the start of heat exposure until T(co) increased to 42 degrees C (the onset of heatstroke), but there was not significantly difference between the two groups; (4) The time of the onset of heatstroke (TOHS) and survival time (ST) of AG group were significantly longer than those of control group; (5) The plasma NO concentrations of the two groups were significantly higher at 60 min than at 0 min after the start of heat exposure, and the plasma NO concentrations of control group were significantly higher than that of AG group at 60 min after the start of heat exposure.</p><p><b>CONCLUSION</b>iNOS may contribute to heatstroke, and aminoguanidine can provide protective effects on heatstroke as a selective iNOS inhibitor.</p>

Effects of artesunate on CD14 and toll-like receptor 4 in peritoneal macrophages of mice with heat stroke endotoxemia / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To study the effects of artesunate on CD14 and toll-like receptor 4 (TLR 4) expressions in peritoneal macrophages of mice with heat stroke endotoxemia.</p><p><b>METHODS</b>Kunming mice were randomly divided into the normal temperature group, the hyperthermia group, the normal saline (NS) group and the artesunate group (both i.p.60 mg/kg daily for consecutive five days). The normal temperature group was exposed to the condition of dry bulb temperature (Tdb) 25 degrees C +/- 0.5 degrees C and relative humidity (RH) 43% +/- 5% for 2 hours, while other groups were exposed to the condition of Tdb 35 degrees C +/- 0.5 degrees C and RH 65% +/- 5%. The mRNA expressions of CD14 and TLR 4 in peritoneal macrophages and concentrations of tumor necrosis factor alpha (TNF alpha) in plasma were observed in different time points (1 hour and 2 hour).</p><p><b>RESULTS</b>The mRNA expressions of CD14 and TLR 4 in the normal temperature group were 0.34% +/- 0.047% and 0.31% +/- 0.062% respectively. The expressions of two receptors at 1 hour in the hyperthermia group were significantly increased to 0.53% +/- 0.085% and 0.45% +/- 0.049% compared with the normal group and kept increased at 2 hour (P < 0.01). The mRNA expressions at 1 hour in the NS group were significantly increased but a little bit decreased at 2 hour. The mRNA expressions of CD14 and TLR 4 at 1 hour in the artesunate group were 0.26% +/- 0.051% and 0.25% +/- 0.084% respectively and a little bit decreased at 2 hour. The change of TNF-alpha in each group was almost consistent with the changes of CD14 and TLR 4.</p><p><b>CONCLUSION</b>Artesunate can reduce significantly the expressions of CD14 and TLR 4 in LPS signal transduction pathway and the concentration of TNF-alpha, which perhaps is one of the most important mechanisms that artesunate fights against endotoxemia.</p>

Arterial blood gas analysis in Lipopolysaccharide-heat co-stressed rats / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To observe the change in vital signs and arterial blood gas in Lipopolysaccharide (LPS)-injected heat exposed rats.</p><p><b>METHODS</b>Male pathogen-free Wistar rats were randomly assigned to the following groups: saline-injected normothermic control (C-Group), saline-injected heat exposed (H-Group), LPS-injected normothermic control (L-Group), LPS-injected heat exposed (HL-Group). Rectal temperature (Tr), heart rate (HR), mean arterial pressure (MAP), arterial blood gas were continually monitored.</p><p><b>RESULTS</b>(1) The rats in HL-Group displayed significantly high values of Tr (43.04 degrees C +/- 0.11 degrees C) and HR [(660 +/- 42) beats/min] and low values of MAP [(49.0 +/- 3.5) mm Hg] compared with C-Group. There was a significant difference in the values of Tr, HR, and MAP between HL-Group and L-Group and in the values of HR and MAP between HL-Group and H-Group. (2) The values of PaO(2), HCO(3)(-), PaCO(2) were significantly lower than those in C-Group at 40 min after LPS-injected heat stress. At 120 min, the PaO(2) [(11.59 +/- 1.11) kPa], HCO(3)(-) [(10.42 +/- 1.06) mmol/L], PaCO(2) [(2.82 +/- 0.81) kPa] in HL-Group were significantly lower than those in L-Group. A significant difference in the values of HCO(3)(-) and PaCO(2) between HL-Group and H-Group was also observed.</p><p><b>CONCLUSION</b>LPS-injected heat stress primes the rat to advance and augment the change in vital signs, arterial blood gas, and systemic inflammatory response syndrome.</p>

Protective and anti-fatigue effects of aspirin against heatstroke in rats / 生理学报

The purpose of this study is to determine whether aspirin can reduce interleukin-1beta(IL-1beta) concentration and exert protective effects against heatstroke. The heatstroke rat model was established through exposing rat to a high ambient temperature (HAT, Ta 41 degrees C, relative humidity 65%) in a simulative HAT chamber to induce heatstroke. Three parts were performed in the present experiment: (1) To determine the effects of pretreatment with aspirin against heatstroke;(2) To prove the effects of specifically reducing inducible nitric oxide synthase (iNOS) against rat heatstroke by iNOS selective prohibitor aminoguanidine (AG);(3) To determine the effects of aspirin against heatstroke and fatigue. In part 1 and 2, Sprague-Dawley rats were randomly assigned to control and aspirin groups or AG groups respectively. Mean arterial blood pressure (MAP), colonic temperature (T(co)), electrocardiograph (ECG) were monitored during heat exposure (HE) and blood samples were taken 0 and 60 min after HE for IL-1betaassay or nitric oxide (NO) assay. In part 3, additional control and aspirin groups of conscious rats were put in a barrel with 41 degrees C water and kept swimming until drowning over 10 s, and then intervals were recorded as survival time. The results from part 1 showed that from 0 to 50 min after HE, MAPs of control group and aspirin group were not significantly different. About 50-60 min after HE, MAPs of both groups were decreased abruptly and MAPs of control group were decreased significantly in comparison with those of aspirin group. T(co) of both groups was increased until to 42 degrees C, without significant difference. Time of heatstroke onset was not significantly different, while survival time was significantly longer in aspirin group than that in control group. Plasma IL-1betaconcentrations in both groups were significantly increased after HE, and the concentration was significantly higher in the control group than that in aspirin group 60 min after HE. In part 3, the survival time was significantly longer in aspirin group than that in control group. In part 2, MAPs of both groups from 0 to 50 min after HE were not significantly different, whereas 55-60 min after HE, MAPs of control group were decreased significantly in comparison with those of AG group;T(co) of both groups was increased after HE until to 42 degrees C, but without significant difference. The time of the heatstroke onset and survival time of AG group were significantly longer than that of control group;the plasma NO concentrations of two groups were significantly higher 60 min after HE than those 0 min after HE, and the plasma NO concentration of control group was significantly higher than that of AG group 60 min after HE. In conclusion, IL-1betamay contribute to heatstroke through inducing iNOS, which attenuates the tone of peripheral blood vessel, and pretreatment with aspirin can provide preventive effects against heatstroke and reinforce the heat and fatigue endurance, which may be associated with inhibition of systemic IL-1betalevels and local iNOS levels.

Mechanism of Ca2+ on the hyperthermia-induced apoptosis of rat hippocampal neurons in vitro / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To study the mechanism of Ca(2+) on the apoptosis induced by hyperthermia in neonate rat hippocampal neurons to provide the applicative evidence of dantrolene for preventing brain injuries.</p><p><b>METHODS</b>Dantrolene, Ca(2+) specific blocking agent, was used in the hyperthermia-induced apoptosis of primary hippocampal neurons in vitro to observe its effect on the apoptosis, fluorescent intensity, and dynamic change of Ca(2+) by flowcytometry and laser confocal microscopy.</p><p><b>RESULTS</b>The rate of apoptosis was decreased significantly after hyperthermia treatment by dantrolene sodium. The intracellular Ca(2+) fluorescent intensity in 42 degrees C treatment group (107.35 +/- 6.0) was significantly lower than that in control group (159.12 +/- 33.8). The concentration of Ca(2+) began to decrease 20 approximately 25 s after adding dantrolene sodium, and reached the lowest level about 50 s later, and then kept lower than the basal level.</p><p><b>CONCLUSION</b>Dantrolene sodium has an important protective effect on hippocampal neurons apoptosis induced by hyperthermia and may have some applicative value of preventing heat-induced brain injury.</p>

Protective effects of heat shock response on circulatory collapse induced by hyperthermia / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To investigate the protective effects and mechanism of heat shock response (HSR) on circulatory collapse induced by hyperthermia.</p><p><b>METHODS</b>Two experiments were carried out: (1) Protective effects of HSR. Rats were divided into 2 groups: heat shock (HS) group, sham control (SC) group. After HS group was pretreated with heat shock and recovered for 20 h at room temperature, both groups were exposed to heat till death, and blood pressure, electrocardiogram were measured continuously during exposure. Mean arterial pressure (MAP), survival time etc were acquired through Chart software. (2) Mechanism of effects. Rats were divided into 3 groups: HS group, SC group and normal control (NC) group. The treatment in HS and SC groups was identical with that in the first experiment, but it would be terminated at 73 min after heat exposure. Systolic pressure (Ps), diastolic pressure (Pd) etc were recorded and content of NO and HSP70 in myocardium were measured.</p><p><b>RESULTS</b>(1) The survival time in HS group [(102.3 +/- 11.4) min] was longer than that in SC group [(87.9 +/- 7.7) min] and shock revealed later (P < 0.01); (2) During early heat exposure MAP in HS group was not different from that in SC group, but after 60 min MAP in HS group were higher than that in SC group; (3) MAP, Ps, Pd, HR and HSP70 in HS group were significantly higher but content of NO was lower than those in SC group (P < 0.01, P < 0.05).</p><p><b>CONCLUSION</b>HSR may induce upregulation of HSP70 and inhibit excessive production of NO in myocardium, thus result in relief of circulatory collapse induced by hyperthermia.</p>

Effect of hyperthermia combined with trauma on serum nitric oxide and mean arterial pressure in rabbits / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To study the early change of serum nitric oxide (NO) after acute heat exposure with trauma and the effect of NO on mean arterial pressure (MAP), thus to provide theoretical basis for studying the mechanism of NO effect in acute stress.</p><p><b>METHODS</b>The rabbit model of acute heat exposure combined with trauma was established. The animals were divided into four groups, including control, trauma, hyperthermia and hyperthermia combined with trauma. The levels of NO were measured at different time points: 0 h, 1 h, 2 h and MAP was monitored throughout the whole experiment.</p><p><b>RESULTS</b>The concentration of NO declined at first and then increased at 1 h or so after acute heat exposure and trauma. The levels of NO in hyperthermia with trauma group at 1 h, 2 h were (42.75 +/- 8.24), (59.54 +/- 9.05) micro mol/L respectively (P < 0.05), while those in control group were (56.63 +/- 3.79) and (55.22 +/- 7.15) micro mol/L, the difference at 1h between two groups was significant (P < 0.05). Under the circumstance of hyperthermia and trauma, the level of MAP declined to the lowest point at 60 - 70 min and then showed a transient rise, after that, the level declined rapidly.</p><p><b>CONCLUSIONS</b>At the early stage of acute heat exposure and trauma, the concentration of serum NO declined at first and then increased, and had certain relationship with the change of MAP.</p>