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To investigate effects of α-asarone and β-asarone on induced PC12 cell injury and related mechanisms. Aβ toxic injury cell model was induced by Aβ in PC12 cells. PC12 cells were divided into blank control group, model control group, α-asarone group (0.5, 1.0, β-asarone group (6.3, 12.5, vasoactive intestinal peptide (VIP) group, and VIP antagonist control group. Cell survival rate was detected by CCK-8 kit; cell apoptosis rate was detected by flow cytometry. The levels of inflammatory cytokines interleukin (IL)-1, , tumor necrosis factor (TNF)-α, oxidation-related inducible nitric oxide synthase (iNOS), nitric oxide (NO), apoptosis factors caspase-3 and p53 were detected by ELISA method. The expressions of C-Jun N-terminal kinase (JNK) and p38 mitogen-activated protein kinase (p38MAPK) were detected by Western blotting. Compared with model control group, cell survival rates of group, β-asarone group and VIP group increased; the cell apoptosis rate decreased; levels of apoptosis-related factors caspase-3, p53, inflammatory factors IL-1, TNF-α decreased; IL-10 level increased; levels of oxidization-related factors iNOS and NO decreased; the expression of JNK and p38MAPK protein decreased (all 0.05). α-asarone and β-asarone have protective effects on PC12 cell injury induced by Aβ. β-asarone may inhibit inflammatory factors and oxidation-related factors through promoting VIP secretion, regulating JNK/MAPK pathway, and reducing PC12 cell apoptosis; however, the effect of α-asarone may be not related to VIP secretion.
Assuntos
Animais , Ratos , Derivados de Alilbenzenos , Anisóis/farmacologia , Apoptose , Células PC12RESUMO
Objective To investigate the therapeutic action and molecular mechanisms of Oxalis comiculata L. extracts on rats with alcoholic liver disease. Methods Forty-two male Sprague Dawley(SD)rats were randomly divided into normal control group(n=10),model group(n=8),moderate dose oxalis group(n=8),high dose oxalis group(n=8)and prednisone group(n=8). The model of rat with alcoholic liver disease was established by liquor gavage;after 12 weeks,moderate dose oxalis group,high dose oxalis group and prednisone group were given the total extract of oxalis 3.5 g?kg-1?d-1,7 g?kg-1?d-1 or prednisolone acetate 0.9 mg?kg-1?d-1,respectively,the remaining two groups were given the same amount of normal saline by gavage daily for 6 weeks. Levels of indexes of liver function,superoxide dismutase(SOD),malondialdehyde(MDA),antidiuretic hormone(ADH)and aldehyde dehydrogenase(ALDH)in rats were detected. The pathological changes of liver tissues in rats were observed under light microscope;tumor necrosis factor-α(TNF-α) expression level was detected by immunohistochemical staining. Results Compared with the normal control group, the levels of aspartate aminotransferase (AST), alanine aminotransferase(ALT),alkaline phosphatase(AKP),γ-glutamyl transferase(GGT),MDA were increased significantly,while the levels of SOD,ADH and ALDH were obviously reduced in model group. Compared with the model group,AST,ALT,AKP,GGT and MDA contents were decreased significantly,while the levels of SOD, ADH and ALDH were markedly increased in the drug groups,and the changes of levels of AST,ALT,AKP,GGT, SOD,ADH in high dose oxalis group were the most obvious〔AST(U/L):117.38±22.75 vs. 201.62±17.95,ALT (U/L):33.51±11.64 vs. 59.14±9.52,AKP(U/L):95.19±24.85 vs. 169.39±37.21,GGT(U/L):46.54±14.55 vs. 89.37±12.49,SOD(U/mg):137.03±12.03 vs. 80.64±13.45,ADH(U/L):3.48±0.71 vs. 2.05±0.91,P<0.05 or P<0.01〕,and the most significant changes of MDA and ALDH were in oxalis moderate dose group〔MDA (mmol/mg):2.05±0.64 vs. 3.17±0.61,ALDH(U/L):7.59±1.95 vs. 5.71±1.33,both P<0.05〕. In normal control group,no obvious lesion was seen in the rat liver tissues. In the model group,fatty degeneration of liver cells with formation of bullae was found,while in the moderate and high dose oxalis groups,cells with macrovesicular steatosis were significantly decreased. Immunohistochemical staining showed that the expression of TNF-α in the cytoplasm and part of cell membrane of macrophage was significantly decreased in liver tissues in oxalis moderate and high dose groups. Conclusion These results show that the Oxalis comiculata L. extracts possess certain therapeutic effect on alcoholic liver disease.
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Objective To investigate effects of Ginkgo biloba extract(EGb) on expression of CREB and pCREB in cortex of aging rats.Methods Male Wistar rats were divided into three groups:young control group,old control group and EGb group.Rats in EGb group were treated with intragastric administration of EGb,while rats in the other two groups were treated with distilled water.The spatial learning and memory were evaluated by Morris water maze,and the expression of CREB,pCREB were detected by western blot.Results( 1 ) Compared with young control group (9.6 ± 2.88,41.55 ± 6.30),the swimming time and times through platform in the target quardrant of rats in old control group(6.8 ± 2.49,34.92 ± 4.56) were reduced (P ＜ 0.05 ).The times passing through the platform and the time exploring the target quadrant were more and longer in EGb group(9.4 ± 2.63,41.0 ± 6.68 ) than those in old control group(P ＜ 0.05 ).(2)Compared with rats in young control group( 1.07 ±0.33,0.26 ± 0.04),relative contents of CREB and p-CREB proteins in cortex (0.70 ± 0.21,0.13 ± 0.05 ) weredecreased in old control group(P＜0.05 ).CREB and p-CREB Levels were higher in EGb group ( 1.02 ±0.18,0.18 ±0.02)than those in old control group(P ＜ 0.05 ).Conclusion EGb can ameliorate spatial learning and memory of rats by increasing the expression of CREB and p-CREB in cortex.
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BACKGROUND: Naomaitong injection is a Chinese herbal compound preparation for treatment of ischemic cerebral vascular disease, acting on resisting calcium overload, regulating the imbalance between thromboxane (TXA) and prostaglandin (PG) and blocking lipid peroxidation mediated by free radical so as to protect cerebrum.OBJECTIVE: To observe the effects of naomaitong injection on water and Ca2+ contents, activity of superoxide dismutase (SOD), levels of lipid peroxide (LPO), 6-keto-PG lα and TXA, and compare it with danshen injection.DESIGN: Randomized control experiment was designed.SETTING: Experimental Center of Chengdu University of Traditional Chinese Medicine.MATERIALS: The experiment was performed in Experimental Center of Chengdu University of Traditional Chinese Medicine from October 1997 to February 1998, in which, 72 healthy male Wistar rats were employed, rangroup: Abdominal injection was done with physiological saline 1.67 mL/kg,group): Abdominal injection was done with compound danshen injection groups (naomaitong No.1, No.2 and No.3 groups): abdominal injection was done with naomaitong injection 3.33, 1.67, 0.84 mL/kg successively,twice/day.METHODS: Totally 48 hours after medication, under anesthetized state,the rats in every group were sacrificed to collect brain tissue. The two hemispheres were cut into two pieces from the middle. One of them was prepared into brain tissue homogenate at low temperature. Radioimmunologic analysis method was used to measure 6-keto-PG 1o and TXA B2 levels so as to evaluate the balance between PG and TXA systems. The modified pyorgallol autoxidation and thiobarbituric acid (TBA) colorimetric method were applied to determine SOD activity and LPO level respectively so as to evaluate lipid peroxidation mediated by free radical. The dry and wet weights of other piece were weighed immediately on electronic scale and the water content of brain tissue was calculated to evaluate brain edema. Atomic absorption spectrophotometry (AAS) was used to determine Ca2+content in brain tissue so as to evaluate calcium overload.tissue of rats in every group.content in brain tissue of rats in every group: That in model group was higher remarkably than normal group [(82.27±1.32)%, (77.24±1.36)%;(267.47±15.69), (37.55±13.23) μg/g, P ＜ 0.01]. The water contents in 4 treatment groups were decreased of various degrees. The effect in No.1 group was the strongest [(78.74±1.41)%] and that in danshen group was the weakest [(81.45±1.52)%]. Ca2+ content in danshen group was decreased of various degrees, indicating dose-effect dependence, but, which was near to ty and LPO level in brain tissue of rats in every group: SOD activity in model group was lower remarkably than normal group [(86.18 ±3.17),(131.86±4.67) μkat/g, P ＜ 0.01]. After treated with naomaitong of 3 dosages, that was all improved, indicating dose-effect dependence (P ＜ 0.01). The effect of No.1 group was the strongest [(119.02±4.00) μkat/g],SOD activity in danshen group was near to model group (P ＞ 0.05). LPO level in model group was higher than normal group [(52.46 ±3.25),(32.29±2.23) μmol/L, P ＜ 0.01]. LPO level of every treatment group was lower significantly than model group and the therapeutic effects of No. 1, 2,3 groups were superior to danshen group [(35.68±2.86), (41.54±2.47),1α and TXA B2 in brain tissue of rats in every group: Content of 6-ketoPG 1α in brain tissue of model group was lower remarkably than normal group (P ＜ 0.01). That was improved in all of 4 treatment groups, in which,the therapeutic effects of No.1, 2, 3 groups were superior to danshen group [(43.84±2.98), (35.01±4.32), (29.97±3.81), (22.89±3.64) ng/g, P ＜ 0.01].TXA B2 content in brain tissue of model group was higher remarkably than normal group (P ＜ 0.01). After treatment, the 4 treatment groups lowed significantly TXA B2 content in brain tissue compared with model group,indicating dose difference. That in danshen group was lower than No. 1, 2,3 groups [(40.58±1.34), (32.85±1.43), (34.31±1.39), (37.27±1.52) ng/g, P ＜0.01].CONCLUSION: Naomaitong injection alleviates brain edema, resists calcium ion, regulates imbalance between TXA and PG systems, improves activity of anti-oxidase and is against injury of free radical so as to protect the structure of brain tissue and achieve therapeutic effects, indicating a certain dose-effect relationship. The effect of naomaitong injection is superior to that of compound danshen injection.
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Objective To investigate the therapeutic mechanism of Chinese medicine of Yi Xin Le oral liquid (YXL) on insomnia. Methods Mice spontaneous activities,mice sleep time induced by sodium pentobarbital in the threshold dose and under the threshold dose and mice convulsion induced by strychnine nitrate were observed to evaluate the sedative, hypnotic and anticonvulsive actions of YXL. Results YXL could restrain the spontaneous movement in mice,had a synergistic action with sodium pentobarbital on mice sleep,and could counteract the convulsive attack induced by strychnine nitrate in a dose-dependent manner. Conclusion YXL has obvious sedative, hypnotic and anticonvulsive actions and this will supply evidence for its clinical usage.