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??OBJECTIVE To observe the neuro-protective effects of harpagide on acute cerebral ischemic injury in mice and its mechanism involving mitochondria. METHODS Acute cerebral ischemia were achieved by operation of MCAO in the left brain, random allocation was taken to divide ICR mice into sham group, model group, nimodipine group and harpagide(5, 10, 15 mg??kg^-1) groups. Mice were intraperitoneal injected harpagide immediately after surgery. Nerve function score, content of brain water, brain index and the common changes of brain pathological structure in HE staining were measured; Ability of mitochondria?? Ca²⁺-Mg²⁺-ATPase and protein expression level of caspase-3 in the MCAO mice?? brains was determined; Ultrastructure change of mitochondria under the TEM was observed. RESULTS Compared with model group, the harpagide groups could decreased the nerve function score, the content of brain water, brain index and the volume of ischemia in mice with different degrees in MCAO mice(P<0.05,P<0.01); 10 mg??kg^-1 of harpagide could increased the activity of Ca²⁺-Mg²⁺-ATPase obviously(P<0.01); And significantly decreased the protein expression level of caspase-3(P<0.01); harpagide groups could protect the pathogeny structure and the ultrastructure of mitochondria with different degrees in MCAO mice, decrease edema of mitochondria obviously. CONCLUSION Harpagide could obviously protect acute cerebral ischemia in mice, its therapeutical effects are approached to protecting the activity of brain mitochondria and decreasing protein expression level of caspase-3.     

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??OBJECTIVE To compare the chemical composition and therapeutic effect between Prunella vulgaris L stem leaf and ear, thus to provide evidence for judging whether the stem leaf of Prunella vulgaris L can substitute the ear as an herbal medicine. METHODS Aqueous extracts of the stem leaf and ear of Prunella vulgaris L from different producing areas were analyzed with HPLC-ESI-MSⁿ. The anti-inflammatory effects were observed by inflammatory models of ear edema induced by dimethylbenzene in mice and hind paw edema induced by carrageenan in rats.Enzyme-linked immunosorbent assay (ELISA) was used to detect the plasma level of TNF-?? and antioxidant activities were detected by ABTS method. RESULTS Prunella vulgaris L stem leaf and ear were not significantly different in chemical composition, both of which contained mainly triterpenoids, flavonoids, phenolic acids and other substances. Compared with the model group, Prunella vulgaris L ear significantly reduced the hind paw edema in rats induced by carrageenan from 1 h after oral administration (P<0.05), while the onset time of stem leaf was later than 1 h.Both groups could significantly reduce the ear edema in mice induced by dimethylbenzene(P<0.01). The TNF-?? levels in the Prunella vulgaris L stem leaf and ear groups [(24.16??1.24) and (24.33??2.36 )ng??mL^-1] were lower than that in the model group [(31.34??1.94) ng??mL^-1] (P<0.01).Prunella vulgaris L stem leaf and ear groups showed strong antioxidant activities in the ABTS^??+ scavenging test. CONCLUSION The contents of the main constituents in Prunella vulgaris L stem leaf and ear have significance differences.The RESULTS of animal tests indicate that the aqueous extracts of Prunella vulgaris L stem leaf and ear have significant anti-inflammatory and antioxidant effects.     

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??OBJECTIVE To investigate the effect of ??-secretase inhibitor DAPT in inflammation-induced brain white matter injury in neonatal mice.METHODS Sixty C57BL/10J neonatal mice are randomly divided into control group, control+DAPT (10 mg??kg^-1) group, inflammation (LPS) group, LPS+DAPT group (inflammation exposure after 10 mg??kg^-1 DAPT treatment). All neonatal mice were killed and brain was removed to the following observation and detection:at P5, the mRNA expression variation of IL-1??, IL-8,TNF-??,Hes1 and NICD by Real-time PCR methods. Oligodendrocytes were identified by immunofluorescence staining. Myelin basic protein (MBP) protein expression was detected by Western blot assay.RESULTS LPS group showed brain injury characterized by inhibition of brain development. There were significant differences in mRNA expression of IL-1??, IL-8, TNF-??, Hes1 and NICD between LPS+DAPT group and LPS group (P<0.05), and the mRNA expression of IL-1??, IL-8,TNF-??,Hes1 and NICD in inflammation-treated were significantly increased than control group (P<0.05). The results showed more expression of MBP in LPS+DAPT group compared with LPS group (P<0.05). Compared with the blank control group, which was obviously decreased after 48 h of inflammation (P<0.05).CONCLUSION Inflammation leads to abnormal of notch signal expression in neonatal mice, and which is shows inflammation involved in brain damage.Its mechanism is probably associated with the maturation of oligodendrocytes.     

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??OBJECTIVE To investigate the protective effect and possible mechanism of Rhizoma Coptis(RC) on lipopolysaccharide(LPS)-induced inflammatory injury in rat hepatocytes(BRL). METHODS LPS-induced BRL cells injury model was established in vitro, then the damaged cells were given different interventions and treatment with 0.175, 0.1 mg?? mL^-1 RC aqueous extract as the test drug, and dexamethasone(Dex) as positive control drug. The optimal test doses of LPS and RC aqueous extract were selected and determined by cell counting kit-8(CCK-8), the cellular apoptosis rate was determined by flow cytometry, TLR4/NF-??B and TLR4/IRF3 signaling pathways and the mRNA level of related inflammatory mediators(TNF-??, IL-1??, IL-6) were detected by RT-PCR, the NF-??B p65 protein expression was analysed by Western blot and immunofluorescence techniques. RESULTS ??Compared with normal control group, 0.1 mg??mL^-1 LPS affected on BRL cells for 24 h, the cell survival rate was decreased significantly(P<0.01), the apoptotic rate increased significantly(P<0.01), the mRNA level of TLR4, NF-??B, IRF3, TNF-??, IL-1??, IL-6 were significantly increased(P<0.01), and the NF-??B p65 protein expression was increased. ??Compared with the model group, 0.1 and 0.175 mg??mL^-1 RC affected on LPS-induced BRL cells for 24 h, the survival rate of BRL cells was increased significantly(P<0.05), the apoptotic rate decreased significantly(P<0.01), the mRNA level of TLR4, NF-??B, IRF3, TNF-??, IL-1??, IL-6 and the NF-??B p65 protein expression were decreased significantly(P<0.01). CONCLUSION Rhizoma Coptis has obviously protective effect on LPS-induced inflammatory injury in rat hepatocytes(BRL), the mechanism of which may be related with inhibiting apoptosis, reducing the release of inflammatory factors such as TNF-????IL-1?? and IL-6, blocking NF-??B p65 protein nuclear translocation, interfering the R4/NF-??B and TLR4/IRF3 signaling pathway.     

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??OBJECTIVE To study the effects of ganoderma spore oil(GSO) on behavior of the mice with chronic unpredictable mild stress (CUMS) and its possible neurophysiology mechanisms. METHODS Thirteen different kinds of chronic unpredictable mild stress were given to the male BALB/C mice for establishing the mouse model of depression. The mice were treated with GSO at 3 doses (850, 283, 141.5 mg??kg^{- 1}??d^-1) or vehicle [(oil) or fluoxetine (10 mg??kg^{- 1}??d^-1)] by oral administration from the 3rd week. After 2 weeks administration, the mice was evaluated by behavioral tests, and the contents of hippocampal glutamate (GLU) and ??-amino butyric acid (GABA) were analyzed by reversed phase high performance liquid chromatography (RP-HPLC). The contents of hippocampal brain derived neurotrophic factor (BDNF) were measured by ELISA kit. RESULTS Compared with model group, GSO increased the body weight, sucrose preference rate and open field test score, shortened the immobility time in the tails suspension test and forced swimming test in the depression mice (P<0.05 or P<0.01). Meanwhile, GSO significantly decreased the contents of GLU (P < 0.01 ) and increased the contents of BDNF(P<0.01), and the contents of GABA did not changes (P>0.05) in the hippocampus. CONCLUSION GSO shows obvious anti-depressant effect on depressant model mice. The antidepressant effect of GSO may be related to decreasing GLU contents and increasing BDNF contents.     

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??OBJECTIVE To investigate the anti-inflammatory effects of extract of Scutellaria baicalensis Georgi (SGE) and underlying mechanism by using LPS-induced microglial BV2 cells. METHODS MTT assay was used to observe the cell viability. The content of NO in cell supernatant was measured by Griess reagent. The levels of IL-1??, IL-6 and TNF-?? were detected by ELISA kits. The intracellular TLR4 expression was assayed by Western blotting. RESULTS The levels of NO, IL-1??, IL-6 and TNF-?? were significantly increased induced by LPS in the supernatant of BV2 cells (all P<0.01). However, co-treatment with SGE 100 ??g??mL^-1 significantly decreased the production of related inflammatory factors including NO (P<0.01), IL-1??(P<0.01), IL-6 (P<0.01) and TNF-?? (P<0.05). Furthermore, SGE significantly inhibited the TLR4 expression induced by LPS in BV2 cells. CONCLUSION SGE is able to alleviate LPS-induced inflammatory responses in BV2 cells through down-regulation of TLR4 protein expression suggesting that SGE has therapeutic potential for the treatment of neuroinflammatory diseases.     

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??OBJECTIVE To observe whether low concentration (1??10^-8 mol??L^-1) of ouabain (OUA)can increase the contractility in rat cardiocytes and investigate the Na/K pump signal transduction pathways related to positive inotropic action following the low concentration of OUA. METHODS On enzymatic isolation of rats ventricular myocytes, the Na⁺/K⁺ pump current (Ip) was by whole-cell patch-clamp, in order to observe the low concentration of OUA on Ip. The contraction of a single myocyte was assessed by a video-based motion edge-detection system. ??To detect and compare the potentiations of 1??10^-8-1??10^-3 mol??L^-1 OUA on the contractility in rat cardiocytes. ??The cardiocytes were pre-treated with PP2(1 ??mol??L^-1), NAC(100 ??mol??L^-1), PD98059(50 ??mol??L^-1)for 5 min, and the effects of the signals transduction inhibitors on the positive inotropic effect of 1??10^-8 mol??L^-1 OUA was recorded. RESULTS The 1??10^-8-1??10^-3 mol??L^-1 OUA increased the contractility of rat cardiocytes (P<0.01). Compared 1 ??mol??L^-1 PP2 +1??10^-8 mol??L^-1 OUA group with 1??10^-8 mol??L^-1 OUA group, the contraction amplitude decreased (P<0.05), and compared 100 ??mol??L^-1 NAC +1??10^-8 mol??L^-1 OUA group with 1??10^-8 mol??L^-1 OUA group, the contraction amplitude also decreased (P<0.01), no significant difference in contraction amplitudes between 50 ??mol??L^-1 PD98059+1??10^-8 mol??L^-1 OUA group and 1??10^-8 mol??L^-1 OUA group (P>0.05). CONCLUSION The 1??10^-8-1??10^-3 mol??L^-1 OUA could increase the contraction amplitude of cardiocytes in rats in concentration-dependent manner. Positive inotropic effect of OUA in low concentration is related to Na/K pump signal transduction. Multiple signal pathways regulate the positive inotropic effect of 1??10^-8 mol??L^-1 of OUA, including the Src/ROS signal pathway.     

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??OBJECTIVE To investigate the effect of acute exposure to 4 300 m altitude environments on the body pathophysiological, serum, TNF-?? and IL-1?? of Wistar rats and protective effect of methazolamide on Wistar rats. METHODS Twenty-eight Healthy adult Wistar rats were randomly divided into plain(altitude of 55 m) control group, high altitude(altitude of 4 300 m) model group, high altitude methazolamide group, and high altitude acetazolamide group. After being intragastric administration with 0.9% sodium chloride injection, methazolamide(2 times a day, 2.23 mg??kg^-1) and acetazolamide(2 times a day, 22.33 mg??kg^-1) for 5 consecutive days. The biochemical, blood gas, the pathological results of rats were analyzed. The TNF-?? and IL-1?? content were detected from the blood samples. RESULTS Blood and biochemical results showed the high altitude might cause dehydration in rats. Compared with the plain control group, each index of the high altitude model group changed significantly(P??0.01), compared with the high altitude group, the aspartate transaminase(AST), alanine aminotransferase(ALT), pH value, bicarbonate concentration(cHCO₃^-), buffer base(BB), base excess(BE) of methazolamide and acetazolamide group were significantly decreased(P??0.01), indicated that methazolamide and acetazolamide had protective effect on rat liver. The total protein(TP), urea solution(UREA), partial pressure of carbon dioxide(PaCO₂), sodium concentration(cNa⁺), chloride concentration(cCl^-) were significantly increased(P??0.01), indicated that the high altitude group had metabolic acidosis and respiratory alkalosis, and liver and lung tissue had pathological damaged. Compared with the acetazolamide group, the methazolamide group damaged less. Compared with plain control group, serum TNF-?? of high altitude groups significantly increased, IL-1?? of high altitude groups decreased significantly, which, serum TNF-??, IL-1?? levels of acetazolamide and methazolamide group were significantly higher than high altitude model group(P<0.01). CONCLUSION Methazolamide can improve acute high altitude physiological and biochemical status of rats, reduce inflammatory injury, with a good protective effect of hypoxia.     

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??OBJECTIVE To investigate the effect of genipin on hypoxia/reoxygenation induced oxidative stress injury in H9c2 cell line. METHODS For mimicking myocardial ischemia/reperfusion (I/R) injury, H9c2 cells were cultured in vitro and established hypoxia/reoxygenation induced oxidative stress injury model. H9c2 cells were divided into six groups: control group, H/R group, H/R+0.5 ??mol??L^-1genipin group, H/R+1.25 ??mol??L^-1genipin group, H/R+2.5 ??mol??L^-1genipin group and H/R+10 ??mol??L^-1genipin group. Cell viability was detected by cell counting kit-8 assay (CCK-8). The levels of lactate dehydrogenase (LDH), intracellular total superoxide dismutase (T-SOD) and malondialdehyde (MDA) were assessed using microplate reader. Confocal laser scanning microscope was performed to examine the production of reactive oxygen species (ROS) and the level of mitochondrial calcium (m) as well as the depolarization ratio of mitochondrial membranes potential(????m). The protein expression of cytochrome-C was detected by Western-blot. RESULTS Comparing to control group, the cell viability of H/R group was significantly decreased in a time-dependent manner(r=-0.82,P<0.01). Low concentration(1.25-40 ??mol??L^-1)of genipin could improve cell viability exposed to H/R treatment (P<0.05), on the contrary, high concentration(80-320 ??mol??L^-1) of genipin remarkably reduced the cell viability (P<0.05). In compared with H/R group, the levels of LDH release, MDA production and ROS production, the level of m, depolarization ratio of ????m and the protein expression of cytochrome-c in H/R+(1.25-10 ??mol??L^-1) genipin groups were notably lessened, while the level of T-SOD was increased(P<0.01 or P<0.05). CONCLUSION Genipin could reduce H/R-induced oxidative stress injury, the mechanism might be connected with balancing the oxidative stress products and anti-oxidation enzyme system as well as improving mitochondrial dysfunction.