Arsenic trioxide regulates the production and activities of matrix metalloproteinases-1, -2, and -9 in fibroblasts and THP-1 / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>The elevated matrix metalloproteinase (MMP) activity is an important cause of chronic wound healing failure. Arsenolite, whose main component is arsenic trioxide (As2O3), is a common traditional Chinese medicine wildly used in treating chronic wounds; it can remove necrotic tissue and promote tissue regeneration. This research was designed to evaluate the effects of As2O3 on production and activities of MMP-1, MMP-2 and MMP-9, and on regulation of its signal transduction pathway in human skin fibroblasts (HSFb) and human monocyte line (THP-1 cells) that were in an inflammatory state.</p><p><b>METHODS</b>We established three cell models; HSFb activated by TNF-α, THP-1 cells activated by phorbol 12-myristate 13-acetate (PMA) and an HSFb-THP-1 co-culture system. Three cell models was cultured with As2O3 for 24 hours. The levels of MMP-1, MMP-2, MMP-9, TNF-α and IL-1β in the cell culture supernatants were assayed by ELISA. The mRNA expressions of MMP-1, MMP-2 and MMP-9 were determined by RT-PCR. The activities of MMP-2 and MMP-9 were tested by Gelatin zymography assays. The phosphorylation levels of ERK1/2 and p38MAPK were assayed by Western blotting.</p><p><b>RESULTS</b>As2O3 inhibited the expression of MMP-1, MMP-2 and MMP-9 mRNA, the secretion and activity of MMP-1, MMP-2 and MMP-9 in HSFb and THP-1 cells in the inflammatory state (P < 0.05 and P < 0.01 respectively). It also inhibited the secretion of TNF-α and IL-1β in THP-1 cells and in the co-culture system (P < 0.05 and P < 0.01, respectively). It also decreased the phosphorylation of ERK1/2 and p38 MAPK in HSFb and THP-1 cells (P < 0.05 and P < 0.01, respectively).</p><p><b>CONCLUSIONS</b>As2O3, as a main component of arsenolite, can inhibit the production of MMPs by HSFb and THP-1 cells in an inflammatory state through inhibiting the release of inflammatory factors and the activation of the MAPK cascade pathway. This may be a possible mechanism for arsenolite healing chronic wounds.</p>

Neuroprotective effects of salvianolic acid B against oxygen-glucose deprivation/reperfusion damage in primary rat cortical neurons / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Cerebral ischemia-reperfusion injury is the main reason for the loss of neurons in the ischemic cerebrovascular disease. Therefore, to deeply understand its pathogenesis and find a new target is the key issue to be solved. This research aimed to investigate the neuroprotective effects of salvianolic acid B (SalB) against oxygen-glucose deprivation/reperfusion (OGD/RP) damage in primary rat cortical neurons.</p><p><b>METHODS</b>The primary cultures of neonatal Wister rats were randomly divided into the control group, the OGD/RP group and the SalB-treatment group (10 mg/L). The cell model was established by depriving of oxygen and glucose for 3 hours and reperfusion for 3 hours and 24 hours, respectively. The neuron viability was determined by MTT assay. The level of cellular reactive oxygen species (ROS) was detected by fluorescent labeling method and spin trapping technique respectively. The activities of neuronal Mn-superoxide dismutase (Mn-SOD), catalase (CAT) and glutathione peroxidase (GSH-PX) were assayed by chromatometry. The mitochondria membrane potential (ΔΨ(m)) was quantitatively analyzed by flow cytometry. The release rate of cytochrome c was detected by Western blotting. The neuronal ultrastructure was observed by transmission electron microscopy. Statistical significance was evaluated by analysis of variance (ANOVA) followed by Student-Newman-Keuls test.</p><p><b>RESULTS</b>OGD/RP increased the level of cellular ROS, but decreased the cell viability and the activities of Mn-SOD, CAT and GSH-PX; SalB treatment significantly reduced the level of ROS (P < 0.05); and enhanced the cell viability (P < 0.05) and the activities of these antioxidases (P < 0.05). Additionally, OGD/RP induced the fluorescence value of ΔΨ(m) to diminish and the release rate of cytochrome c to rise notably; SalB markedly elevated the level of ΔΨ(m) (P < 0.01) and depressed the release rate of cytochrome c (P < 0.05); it also ameliorated the neuronal morphological injury.</p><p><b>CONCLUSION</b>The neuroprotection of SalB may be attributed to the elimination of ROS and the inhibition of apoptosis.</p>

Regulation function of Qingnao drop pilula to MARCKS mRNA express changes in acute cerebral ischemia hippocampus / 中国中药杂志

<p><b>OBJECTIVE</b>To observe the contribution of Qingnao drop pilula to the alteration of myristoylated alanine-rich C kinase substrate (MARCKS) mRNA expression in acute multi-infarction hippocampus.</p><p><b>METHOD</b>Rat models of acute multi-infarction were established by injecting the embolus of blood powder through the right external carotid arteryinto the internal carotid artery, rats were randomly divided into five groups (n = 12 in each): normal, sham operation, model, Chinese medicine treatment, and Western medicine treatment. Qingnao drop pilula (133.28 mg x kg(-1)), nimodipine (7.25 mg x kg(-1)) were administered respectively to Chinese medicine treatment group and Western medicine treatment group by gavage, equal volume of normal saline were given to three groups. Rats were treated with drugs starting at 3rd day before the operation, one time per day. Observing morphologic changes in hippocampus by optical microscope and electron microscope. Detecting expression level of MARCKS mRNA in hippocampus by semi-quantification PCR method.</p><p><b>RESULT</b>Hippocampus cells arrange tidy, administrative levels were compactness in normal group, which cells differentially impaired in model group, Chinese medicine treatment group and Western medicine treatment group. Hippocampus cells damage of Chinese medicine treatment group have more reckless than the model group in histopathology. The MARCKS mRNA were expressioned in model group vs medication treatment groups, in Chinese medicine treatment group vs the model group.</p><p><b>CONCLUSION</b>Qingnao drop pilula can alleciate histomorphology lesion of hippocampus when occurring acute multi-infarction, to turn slower MARCKS mRNA expression, may play a neuroprotective effect role through accommodating PKC-MARCKS signal transduction system.</p>

Effect of Xinnao Shutong capsule on apoptosis after cerebral ischemia and reperfusion in rats / 中国中药杂志

<p><b>OBJECTIVE</b>To study apoptosis-regulating cytokines and apoptosis on focal cerebral ischemia and reperfusion in rats treated with Xinnao Shutong capsule.</p><p><b>METHOD</b>Rat models of focal cerebral ischemia and reperfusion were established by thread ligation in middle cerebral artery occlusions (MCAO). After 24 hours, the brains were removed to detect changes of protein expression of Bax, Bcl-2, Fas, Fas-L and caspase-3 by immuno-hisochemistry, and apoptosis of cortical neurons by TUNEL RESULT: Compared to control, brain cortex have decreasing the protein expression of Bcl-2 and the ratio of Bcl-2/Bax, increasing the protein expression of Bax, Fas, Fas-L and caspase-3 of ischemia and reperfusion models group (P < 0.01). Xinnao Shutong capsule group could increase the protein expression of Bcl-2 and the ratio of Bcl-2/Bax, and obviously decrease the protein expression of Bax, Fas, Fas-L and caspase-3, then reduce the number of apoptotic cells of cortex (P < 0.01).</p><p><b>CONCLUSION</b>Xinnao Shutong capsule protect injured rat brain tissue, may be related to decrease neuronal apoptosis and adjusted protein expression of apoptosis-regulating cytokines.</p>

Effect of salvianolic acid B on brain energy metabolism and hydrocephalus of cerebral ischemia in mice at different time / 药学学报

Mice pathological model of acute cerebral ischemia was established. In order to observe the effect of salvianolic acid B (Sal B) on brain energy metabolism and hydrocephalus in the brain of mice at different ischemic times, the energy charge (EC), content of phosphocreatine (PCr), level of lactic acid (Lac), activity of Na+ -K+ -ATPase, brain index and water content of brain were measured at 6, 12, 18, 24, and 30 min, separately after ligating bilateral common carotid arteries in mice. NIH mice were randomly divided into sham-operated group (sham), cerebral ischemia group (ischemia), Sal B-treated group (Sal B) and nimodipine-collated group (Nim). At 6 min after cerebral ischemia, EC, content of PCr and activity of Na +-K -ATPase began to decrease, while level of Lac, brain index and water content of brain increased gradually. However, Sal B (22.5 mg x kg(-1) improved pathophysiological changes at different ischemic times. Especially at 30 min after cerebral ischemia in Sal B group, EC (P < 0.01), content of PCr (P < 0.01 and activity of Na+ -K+ -ATPase ( < 0.05) increased significantly. Meanwhile, level of Lac (P < 0.01, brain index (P < 0.01) and water content of brain (P < 0.05) were lower obviously than those of cerebral ischemia group. Sal B could alleviate hydrocephalus by the improvement of energy metabolism in mice with acute cerebral ischemia, that provides scientific evidence that Sal B can be used for the clinical application of ischemic diseases.

Effects of salvianolic acid B on cerebral energy charge and activity of ATPase in mice with cerebral ischemia / 中国中药杂志

<p><b>OBJECTIVE</b>To observe the effect of salvianolic acid B (SalB) on high energy phosphate and activity of ATPase of cerebral ischemia in mice, and to study the role of SalB on hydrocephalus further.</p><p><b>METHOD</b>NIH mice were divided into four groups randomly: Sham-operated group, cerebral ischemia group, SalB-treated group and Nimodipine (Nim)-collated group. In Sal B-treated group, mice were injected with SalB (22.5 mg x kg(-1)) in vena caudalis at 30 min before the experiment. In Nim-collated group, Nim (0.03 mg x kg(-1)) was injected into tail vein at the same time, while the mice in Sham-operated group and cerebral ischemia group were injected the same volume normal saline. The acute cerebral ischemia model was established by ligating bilateral common carotid arteries for 30 min in mice, then the mice were killed and the content of adenosine triphosphate (ATP), adenosine diphosphate (ADP), adenosine monophosphate (AMP), phosphocreatine (PCr) were observed, and the cerebral energy charge (EC) was computed. At the same time, activity of Na(+) -K(+) -ATPase and Ca2(+) -ATPase, content of water in brain tissue were measured.</p><p><b>RESULT</b>Compared with cerebral ischemia group, EC and content of ATP, ADP, PCr in SalB-treated group heightened evidently (P < 0.01). Moreover, activity of Na(+)-K+ ATPase and Ca2+ ATPase in SalB-treated group had a remarkable increase (P < 0.01). But the content of water in brain tissue decreased markedly (P < 0.05).</p><p><b>CONCLUSION</b>The mechanism that SalB can relieve content of water in brain tissue of cerebral ischemia in mice, may be associated with improving the content of high-energy phosphoric acid compounds and enhancing the activity of ATPase.</p>

Protective effects of Xinnao Shutong capsule on acute cerebral ischemic injury of multiple infarcts in rats / 中国中药杂志

<p><b>OBJECTIVE</b>To study the effect of Xinnao Shutong capsule (XNST) on energy metabolism dysfunction, free radical injury and inflammatic factors in the course of acute cerebral ischemic damage, and try to reveal the mechanism of the protection against ischemia.</p><p><b>METHOD</b>60 male Wistar rats weighing 280 - 320 g were randomly divided into five groups: normal, sham operation, model, XNST treatment( XNST-T) , and Western medicine treatment (WM-T) group. Acute multi-infarct model in rats was induced by injecting the embolus of blood powder through the right external carotid artery (ECA) into the internal carotid artery (ICA). At 72 hours after ischemia, morphologic change and the express of tumor necrosis factor-alpha (TNF-alpha) and interleukin -1beta ( IL-1beta) in hippocampus CAl section and cortex were observed, biochemical criterions including the activity of Na+ -K+ -ATPase, lactate dehydrogenase (LDH), superoxide dismutase (SOD), and the content of malondialdehyde (MDA) in hippocampus were examined.</p><p><b>RESULT</b>The morphologic change of hippocampus and cortex in both XNST-T and WM-T groups was milder than that in model group. The activity of Na+ -K+ -ATPase, LDH and SOD in hippocampus were all significantly decreased in model group (P <0. 01), and elevated in XNST group (P <0. 01) as well as in WM-T group (P <0. 01). The content of MDA in hippocampus was significantly increased in model group (P <0. 05), and was reduced in XNST group (P <0. 05) as well as in WM-T group (P <0. 01).</p><p><b>CONCLUSION</b>The results reveal that XNST has the protective effect against cerebral ischemic injury. And its possible mechanism is that XNST can prevent the upper pathological process.</p>

Research of kingsbrain on improving the learning and memory ability of MCAO rats / 中国中药杂志

<p><b>OBJECTIVE</b>To explore the influence of Kingsbrain (GETO) on the learning memory impairment of rats with cerebral ischemia.</p><p><b>METHOD</b>Rats with cerebral ischemia were administered GETO orally once a day for one month. The ability of spatial-learning memory of rats was evaluated by Morris Water Maze (MWM). Duxil was used as a positive control.</p><p><b>RESULT</b>the results of place navigation of MWM showed that at the 3rd time of swimming training, the escape latency of rats of the GETO group, Duxil group and Sham group were shorter than that of model group. The escape latency were (54.1 +/- 43.94), (55.9 +/- 43.49), (50.4 +/- 34.99) and (85.4 +/- 42.8) s, respectively; but there was no significantly difference. After the 6th time of swimming training, the escape latency of rats of the GETO group (37.8 +/- 38.69) s, the Duxil group (37.4 +/- 38.03) s and the sham group (26.9 +/- 21.63) s were significantly shorter than that of model rats (77.5 +/- 47.59) s, P < 0.05, respectively. Comparison of the swimming distance among groups were similar to the escape latency among groups. In the test of spatial probe, results of the ratio of the swimming time of platform quadrant (tP) vs the total swimming time (tT) and the ratio of the swimming distance of platform quadrant (dP) vs the total swimming distance (dT) indicated that the ratios of the GETO group (0.347 +/- 0.0662, 0.344 +/-0.055 1), the Duxil group (0.345 +/- 0.0984, 0.34 +/- 0.0934) and the sham group (0.35 +/- 0.0662, 0.349 +/- 0.0589) were significantly higher than those of the Model group (0.261 +/- 0.0689, 0.274 +/- 0.0544), P < 0.05, respectively.</p><p><b>CONCLUSION</b>GETO can significantly improve the spatial learning and memory ability of rats with cerebral ischemia, which provides the pharmacodynamics evidence for its clinical application of improveing the learning and memory ability in poststroke patients.</p>

Seven emotions generation and five Zang-organs regulation machanism / 中华中医药杂志

Seven emotions generation relates to the interaction between individual and external objective matters,which is accompanied with interaction between internal desire and external matters.The activity of seven emotions bases on essence of zang-fu organs,being regulated by five zang viscera.Heart is the upstream controller,liver is the key organ to maintain normal emotionds,spleen and stomach is the hub of emotional activities,lung is the auxiliary organ, kidney is origin of emotional generation.Five zang viscera's cooperation and interaction generate the seven emotions.

Acupuncture inhibiting responses of spinal dorsal dorsal horn neurons induced by noxious dilation rectum and colon / 中国针灸

<p><b>OBJECTIVE</b>To study on mechanisms of acupuncture in relieving visceral pain.</p><p><b>METHODS</b>In SD rats CRD was used as noxious visceral stimuli. Activities of spinal dorsal horn wide dynamic (WDR) neurons of L1-L13 were recorded by extracellular microelectrode technique. Acupuncture was given at ipsi-lateral and contra-lateral Zusanli (ST 36) of the same segmental innervation of rectum and colon.</p><p><b>RESULTS</b>Visceral noxious afferent could significantly activate spinal dorsal horn convergent neurons, and mechanical stimulation of contra-lateral body surface and hand acupuncture at Zusanli (ST 36) could inhibit this noxious response. When the spinal cord was acutely blocked, the inhibiting CRD effect of needling CRD effect of needling contra-lateral Zusanli (ST 36) completely disappeared.</p><p><b>CONCLUSION</b>Acupuncture and visceral noxious afferent signals converge and interact each other in spinal level, and acupuncture at acupoint can inhibit the spinal dorsal horn neuron respon se activated by visceral noxious afferent and this action needs the participation of the center above the spinal cord.</p>

Effects of Panax notoginseng saponins on rat cardiomyocytes apoptosis induced by angiotengin II in vitro / 中国中药杂志

<p><b>OBJECTIVE</b>To study the protective effects of Panax notoginseng saponins (PNS) on angiotensin II (Ang II)-induced rat cardiomyocyte apoptosis in vitro and the probable mechanism.</p><p><b>METHOD</b>Cultured cardiomyocytes from neonatal rats were stimulated with Ang II. Cell viability was measured by MTT. Apoptosis was evaluated using Acridine Orange (AO) fluorescent dye staining and flow cytometry; Fluo-3 AM was used to test the change of intracellular free calcium.</p><p><b>RESULT</b>It was found that incubating with Ang II (10(-7) mol x L(-1)) for 48 h increased cardiomyocyte apoptosis, PNS (25, 100 mg x mL(-1)) increased myocyte viability. PNS (50 mg x mL(-1)) significantly decreased this Ang II-induced rat cardiomyocyte apoptosis (P < 0.05) and decreased fluorescent intensity of intracellular calcium.</p><p><b>CONCLUSION</b>PNS has a significant effect on Ang II-induced rat cardiomyocytes apoptosis in vitro by alleviating intracellular calcium overload.</p>

Effect of drugs for promoting blood circulation on blood lipids and inflammatory reaction of atherosclerotic plaques in ApoE gene deficiency mice / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To observe the effect of six common Chinese medicinal herbs for promoting blood circulation, including Radix Paeoniae rubra (I), Radix Salviae miltiorrhizae (II), Rhizoma Chuanxiong (III), Radix Notoginseng (IV), Semen Persicae (V) and wine steamed Radix et Rhizoma Rhei (VI), on blood lipids and inflammatory reaction of atherosclerotic plaques in ApoE gene deficiency mice.</p><p><b>METHODS</b>Ninety mice, 6 - 8 weeks old, were divided into 8 groups, the model group, the control group (treated with simvastatin) and the six treated groups treated with the above-mentioned 6 Chinese medicinal herbs respectively. All the mice were fed with the diet of western kind for 13 weeks until the mature atherosclerotic plaques formed in them. Then they were treated with respective drugs for another 13 weeks except those in the model group. All the mice were sacrificed at the end of experiment, their blood was collected for lipids determination, heart and aorta were taken out for determining the level of CD68 in root of aorta, as well as the expressions of monocyte chemotactic protein-1 (MCP-1) and tumor necrosis factor-a (TNF-a) by immunohistochemistry staining.</p><p><b>RESULTS</b>All the 6 Chinese herbs showed regulatory action on blood lipids. The positive expression of CD68 in the model group displayed the highest activity. As compared with the model group, the CD68 positive expressed cells in the control group and the groups treated with Chinese herbs II, III, and IV were lesser (P < 0.05), and the expression of inflammatory factors (MCP-1 and TNF-alpha) in atherosclerotic plaques was significantly lower in the control group and the group treated with Chinese herb VI (P < 0.05).</p><p><b>CONCLUSION</b>Chinese medicinal herbs tested in this study can interfere the maturing progress of atherosclerotic plaques and stabilize the plaques in ApoE deficiency mice, the mechanisms may relate to its actions in regulating lipids metabolism and inhibiting inflammatory reaction. Different Chinese medicinal herbs for activating blood circulation of conventional dosage might show difference in potency and acting links.</p>

Study on the anti-apopotosis induced by hypoxia/hypoglycemia and reoxygenation of panax notoginseng saponins in cultured rat hippocampal neurons / 中国中药杂志

<p><b>OBJECTIVE</b>To study the inhibitory effects of Panax Notoginseng Saponins(PNS) on apoptosis induced by hypoxia/hypoglycemia and reoxygenation in cultured rat hippocampal neurons.</p><p><b>METHOD</b>Apoptosis were measured by flow cytometry, intracellular free calcium concentration([Ca2+]i) was measured with confocal laser scanning microscopy, morphological changes and neuronal necrosis were observed with fluorescence microscope, and meanwhile the leakage of lactic dehydrogenase(LDH) was measured.</p><p><b>RESULT</b>Hypoxia/hypoglycemia cultures for 5 hours and reoxygenation induced neuronal apoptosis and necrosis, and significantly increased neuronal [Ca2+]i and the leakage of LDH. The effects were increased with the extending time of reoxygenation. PNS has could significantly decrease the percentage of neuronal apoptosis and necrosis, and reduce neuronal [Ca2+]i and the leakage of LDH.</p><p><b>CONCLUSION</b>PNS has inhibitory effect on neuronal apoptosis. This effect might be related to its effect of decreasing intracellular free calcium concentration.</p>