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??OBJECTIVE To explore the effects of Dracocephalum total flavonoids(TFDM) on myocardial ischemia/reperfusion injury(MIRI)induced autophagy in rat. METHODS In vivo MIRI model was established with ischemia 30 min and reperfusion 120 min, by ligation of left anterior descending coronary artery. TFDM(30 mg??kg^-1??d^-1) and autophagy activator(1 mg??kg^-1??d^-1) or inhibitor(25 mg??kg^-1??d^-1) pretreatment was given before making the model. The MIRI-induced infarct size, the activities of lactate dehydrogenase(LDH), creatine kinase isoenzyme(CK-MB) in plasma and apoptosis protein caspase-3 in tissue and the expression level of autophagy-related proteins LC3, Beclin-1 were observed. RESULTS Compared with model group, the experiment revealed that autophagy inhibitor alleviated MIRI-induced myocardial damage by decreasing the infarct size, myocardial enzyme LDH and CK-MB leak; and reducing apoptosis protein caspase-3 activity level. In addition, TFDM pretreatment significantly inhibited the expression of autophagy-related proteins LC3 and Beclin-1. CONCLUSION TFDM shows inhibitory effects on MIRI-induced autophagy,which may play an important role in the protection against MIRI.     

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??OBJECTIVE To investigate the expression and purification of protease inhibitor OH-TCI in Escherichia coli and its protective effect and mechanism in myocardial ischemia-reperfusion injury model. METHODS The recombinant plasmid of Pet32a-OH-TCI was transformed into E. coli and induced by IPTG, soluble fusion protein was expressed in the constructed expression system and isolated by a nickel affinity chromatography column. Then, the products was digested by TEV enzyme and was purified by the nickel affinity chromatography column. The adult male rats were divided into two groups randomly:normal saline group and OH-TCI group. At 5 min before the operation, rats were treated with OH-TCI by intraperitoneal injection in the experimental group and was given the same volume of saline in the control group. In all animal experiments, rats left anterior descending coronary artery were ligated for 30 min to construct the model. After 120 min of reperfusion, samples were taken to determine the contents of malondialdehyde (MDA), tumor necrosis factor-?? (TNF-??) and interleukins-6 (IL-6) and the activities of lactic acid dehydrogenase (LDH), creatine kinase (CK) and superoxide dismutase (SOD). Finally, the protective effects of OH-TCI against myocardial ischemia-reperfusion injury in rat model were evaluated. RESULTS The recombinant protease inhibitor OH-TCI was expressed in the E.coli expression system in a soluble pattern and successfully purified by two steps of nickel affinity chromatography. Comparing with those in the control group, the recombinant protease inhibitor OH-TCI could significantly decrease the levels of LDH, MDA and IL-6 (P<0.01)as well as inflammatory factor TNF-?? (P<0.05). CONCLUSION The recombinant protease inhibitor OH-TCI can be expressed successfully in E. coli. Purified recombinant OH-TCI has natural protease inhibitory activity and shows protective effects against ischemia and reperfusion myocardium injury.     

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??OBJECTIVE To study the chemical constituents of different solvent extracts from Fructus Trichosanthis, and to study their antioxidant activity and protection of myocardial ischemia reperfusion injury in rats. METHODS The chemical constituents in different solvent extracts were determined by UV-Vis spectrophotometry. Myocardial ischemia reperfusion model was made by ligation of left coronary artery in rats. Rats were randomly divided into 6 groups, with 6 rats in each group.The sham operation group and the model groups were respectively given normal saline, Fructus Trichosanthis (equivalent to crude drug 22.5 g??kg^-1), compound Danshen dripping pill group(0.085 g??kg^-1).Medicines were given once a day for 7 d. After the last drug 1 h, left coronary artery was ligated for 30 min and then reperfusion was established for 120 min by removing the ligation. During this time, ECG was recorded. At the end, animals were euthanized. Blood was collected to evaluate the contents of CK-MB, MYO-MB, cTnT. The heart was removed and fixed to observe the changes of myocardial tissue by optical microscope. RESULTS Compared with the water extract group and the alcohol extract group of Fructus Trichosanthis, the total amino acids content of Fructus Trichosanthis dichloromethane extract was not detected, but the content of total flavonoids is higher (P<0.01). Compared with the water extraction liquid of Fructus Trichosanthis, the antioxidant properties on DPPH radical of Fructus Trichosanthis dichloromethane extract group is lower (P<0.01), and the antioxidant activity on-OH free radical of the alcohol extract of Fructus Trichosanthis group is lower (P<0.01). Compared with model group, the elevation of ST segment of electrocardiogram was significantly suppressed in each group during reperfusion (P<0.01 or P<0.05). The plasma CK-MB,cTnT and MYO-MB in water extract group were significant lowered (P<0.05). CONCLUSION The extraction of Fructus Trichosanthis is able to decrease the production of oxidants. The water extract of Fructus Trichosanthis could ameliorate myocardial ischemia reperfusion injury.     

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??OBJECTIVE To discuss the effect and mechanism of supplementing qi and nourishing yin(SQNY) decoction on intestinal microbiome of mice with Lewis lung carcinoma after treated with cisplatin. METHODS The model of C57BL/6 mice with lung carcinoma xenograft was established by subcutaneous seeding of Lewis cells and randomly divided into 4 groups (n=16), including saline group (NC group), cisplatin group (DDP group), CAP group (CAP group) and SQNY decoction combined with cisplatin group (DDP+SQNY group). The tumor sizes, weights and intestinal microbiota were detected. The serum levels of IL-6, IL-10, and TNF-?? were detected by ELISA. The dendritic cell (DC) and macrophage subtype in spleen of mice models were detected by flow cytometry. RESULTS ??The mean tumor sizes of mice in DDP group, CAP group, DDP+SQNY groups were all smaller than NC group (P<0.05). And the mean tumor size and weight in DDP+SQNY group were less than the other three groups (P<0.05). The mean animal weights in NC or DDP+SQNY group were higher than those in DDP group and CAP group (P<0.05). ??The mean levels of serum IL-6, IL-10 and TNF-?? in DDP+SQNY group were less than those in the other three groups (P<0.05). ??The mean microbiota richness in DDP+SQNY group were less than those in the other three groups (P<0.05). The proportion of fecal microbiota was significantly changed in DDP+SQNY group, including upregulation of Firmicutes phylum and Bifidobacterium Genus, and downregulation of Bacteroidetes phylum(P<0.05). ??The amount of DC and M1/M2 macrophage subtype in spleen of DDP+SQNY group mice models were more than those in the other three groups (P<0.05). CONCLUSION It??s suggested that SQNY decoction effectively enhances the anticancer effect of cisplatin by increasing the immune function and changing the intestinal microbiota.     

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??OBJECTIVE To investigate the inhibitory effect and possible mechanisms of puerariae isoflavone(PI) on prostatic hyperplasia induced by testosterone propionate.METHODS Forty-eight male Wistar rats were randomly divided into six groups according to their body weight including normal control group, model group, 40, 80, 160 mg??kg^-1??d^-1 PI group, and finasteride positive control group. In addition to the sham operation for rats in the normal control group, the rats in other five groups performed castration surgery. After the restoration, the five groups of rats were subcutaneously injected with testosterone propionate (10 mg??kg^-1??d^-1) for 10 d to establish a benign prostatic hyperplasia model and then the subcutaneous injection was maintained every 2 d. High, middle and low dose PI groups were intragastrically administered (40, 80, 160 mg??kg^-1??d^-1) from the second day when the benign prostatic hyperplasia model was successfully constructed. The positive control group was given finasteride (1.0 mg??kg^-1??d^-1) .Rats in normal and model groups were given an equal volume of saline for 28 d. After the last administration, the prostate and seminal vesicles were separated under anesthesia in rats, the wet weight and volume of the prostate and seminal vesicles were measured. The prostate and seminal vesicles index were calculated too. Rat blood was drawn and dihydrotestosterone(DHT) and estradiol (E2) in the serum were measured. Nitric oxide (NO), nitric oxide synthase (NOS), superoxide dismutase (SOD) and malondialdehyde (MDA) levels in prostate tissues were measured. The prostate tissue in each group was randomly selected for HE staining. The pathological structure of the prostate tissue was observed under an optical microscope.RESULTS Compared with the normal control group, the prostate gland index and seminal vesicle gland index of the model group increased significantly (P<0.01), and the DHT and E2 levels in serum increased significantly (P<0.01). MDA content was increased while NO levels, NOS and SOD activities were significantly decreased (P<0.01). HE staining showed that the size of the prostate gland in the model group was different, there were obvious dilation, hyperplasia and papillary protrusions, and the cavity was full of pink and homogeneous density. The interstitial tissue showed obvious dilations of blood vessels, infiltration of inflammatory cells, and proliferation of fibrous connective tissues. Compared with the model group, the index and volume of prostate and seminal vesicles in the PI and positive control groups were significantly decreased (P<0.05 or P<0.01), and the levels of serum DHT and E2 in the middle and high doses PI groups were significantly lower (P<0.05 or P<0.01). In all treatment groups, MDA content was decreased and NO, NOS, and SOD levels were increased (P<0.05 or P<0.01) except the low-dose PI groups. There was moderately hyperplasia in low-dose PI group, mild prostatic hyperplasia in positive control group and middle-dose PI group, basically no hyperplasia in high-dose PI group.CONCLUSION PI has a certain inhibitory effect on prostate hyperplasia induced by testosterone propionate, especially in the medium and high dose PI groups. The mechanism may be related to the effects of pueraria isoflavone on antioxidant,free radical scavenging in vivo, increasing NOS activity and increasing NO level.     

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??OBJECTIVE To observe the effect of guhong injection(GHI) on tibial fracture healing in rats and to explore the mechanism of the action of GHI. METHODS One hundred and eighty male SD rats were randomly divided into 6 groups with 30 rats in each group: sham operation group, model group, positive drug group(compound ossotide injection, 5 mL??kg^-1), low, medium and high dose of GHI groups(2.5, 5, 10 mL??kg^-1). In addition to the sham operation group, the other groups established the rat model of tibial fracture. All were given once daily intraperitoneal injections and samples were taken at 1st, 2nd, 4th and 6th week. Blood biochemical analysis and Elisa kit detection were performed on blood samples. X-rays, biomechanical tests, immunohistochemistry and RT-PCR were performed on the tibial samples. RESULTS ??After administration for one, two and four weeks, the levels of serum calcium(Ca) and phosphorus(P) in medium and high dose of GHI groups were higher than those in model group(P<0.05). ??X-ray showed that the outer callus growth and the disappearance of fracture line in all dose groups of GHI were faster than those in model group. ??Compared with model group, the maximum load and rigidity of medium and high dose of GHI groups were increased at each time point(P<0.05), and the trend of stress line graph were improved obviously. ??The content of alkaline phosphatase(ALP) in medium and high dose of GHI groups were higher than that in model group at each time point(P<0.05). Compared with model group, the serum levels of PDGF were increased in all dose groups of GHI(P<0.05 or P<0.01). After administration for one, two and four weeks, the serum BMP-2 in all dose groups of GHI were higher than those in model group(P<0.05 or P<0.01). ??Compared with model group, the expression of Runx2 mRNA were increased in medium and high dose of GHI groups, as well as Smad5 protein expression(P<0.05 or P<0.01). CONCLUSION GHI could significantly improve the biomechanical properties of bone in fracture rats. The promotion of fracture healing might be through the upregulation of PDGF and BMP-2 expression in different stages of bone healing, and the regulation of BMP/Smad5/ Runx2 signaling may be one of the mechanisms of promoting fracture healing.     

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??OBJECTIVE To study the effect and action mechanism of the large plant Rhodiola Tablet on myocardial ischemia of rats. METHODS Rat myocardial ischemia models were induced by coronary artery ligation. The levels of AST, ALT, CK and CK-MB in serum were observed, at the same time the expressions of B-raf, Ras and p-ERK1/2 in myocardial tissue were determined. RESULTS The levels of AST, ALT, CK and CK-MB in rat serum, as well as the B-raf, Ras and p-ERK1/2 expression in rats myocardium in the large plant Rhodiola group were significantly lower than that in the model group. CONCLUSION Large plant Rhodiola has a certain antagonism on myocardial ischemia injury in rats caused by coronary artery ligation. The mechanism of action may be related to the decreases in serum AST, ALT, CK and CK-MB,as well as myocardial high expression of the B-raf, Ras and p-ERK1/2.     

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??OBJECTIVE To investigate the ameliorated effect and mechanisms of phenylethanoid glycosides from Pedicularis muscicola Maxim on high altitude memory impairment. METHODS After successfully trained in the 8-arms radial maze, fifty Wistar rats were randomly divided into normoxic control group, hypoxia group, phenylethanoid glycosides 50, 200 and 400 mg??kg^-1 groups(given corresponding dose). Normoxic control and hypoxia groups were administered with distilled water for a week.When drug delivery in the fourth day, hypoxia and phenylethanoid glycosides groups rats were exposed to a simulated of 7 500 m in a specially designed animal decompression chamber. Eight arms radial maze was used to measure spatial memory, HE stained was used to observe the cell morphology in brain tissue and biochemical technique was used to detect the content of MDA and ROS and enzymatic activity of GSH and SOD in brain tissue and serum. RESULTS Compared with the normoxic control group, for hypoxia group rats, WME, RWE and TE were respectively increased by 800%, 71%, and 127.1%(P??0.01) and neuron damage was significantly increased, the enzymatic activity of GSH and SOD were respectively decreased by 60.9% and 18.11%(P<0.05, P<0.01) in brain tissue and plasma while the content of MDA was increased in brain tissueby 74.8% (P<0.01). Compared with the hypoxia group, for phenylethanoid glycosides 200, 400 mg??kg^-1 groups rats, WME,RWE,TE were respectively decreased by 68.44%,63.11%;33.14%,25.34% and 43.91%, 36.72% (P??0.05, P??0.01) and neuron damage was significantly decreased, the enzymatic activity of GSH were respectively increased by 219.76%, 180.75% and 32.81%, 24.10% (P<0.05, P<0.01) and the enzymatic activity of SOD were respectively increased by 9.57%, 13.88% and 15.41%, 15.45% (P<0.05) in brain tissue and plasma,while the content of MDA in plasma were respectively decreased by 42.73%, 42.73% (P<0.01) and MDA and ROS in brain tissue were respectively decreased by 61.71%, 42.79% and 40.76%, 23.53% (P<0.01); for phenylethanoid glycosides 50 mg??kg^-1 group rats, the corresponding indicators had been ameliorated, but there was no significant difference.CONCLUSION Phenylethanoid glycosides of Pedicularis muscicola Maxim can ameliorate high altitude memory impairment, which its involved mechanism may be antioxidant stress and inhibition on cell damage.     

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??OBJECTIVE To observe the effect of Dracocephalum heterophyllum flavonoids(DHBF) of Uygur Medicine on cardiomyocyte hypertrophy, which were induced by angiotensin ??(Ang??), and it could provide a basis for further study to the mechanism. METHODS SD rats,0-3 d of age, neonatal rat myocardial cells cultured in vitro, the experiment was divided into control group, Ang??(1 ??mol??L^-1)group, different concentrations of DHBF(10, 25, 50 ??mol??L^-1)+Ang??(1 ??mol??L^-1) groups, cardiomyocyte hypertrophy were induced by Ang?? 1 ??mol??L^-1 and was intervened using DHBF respectively, CCK-8 method was used to observe the activity of myocardial cells,RT-PCR technique was used to detect the expression of m RNA of cardiac hypertrophy gene atrial natriuretic peptide(ANP) and brain natriuretic peptide(BNP),the internal factor was glyceraldehyde-3-phosphate dehydrogenase(GAPDH) .Confocal laser scanning was used to detect the surface area of myocardial cell was [Ca²⁺]_i;the activity of Ca²⁺-ATP was measured by the enzymatic reaction of fragmentation cell; the concentration of NO and the activity of NOS were determined by colorimetry. RESULTS Compared with the control group, the activity of myocardial cell was(85%??5%) in the Ang??group,which was increased significantly after it was dealed with DHBF and Ang??(P<0.05);RT-PCR results showed the expression of mRNA of ANP and BNP were increased by using Ang??, which were lower by using DHBF and Ang??. The surface area of myocardial cell were increased by using Ang??, which could be reversed by using DHBF and Ang??. Confocal laser scanning showed the concentration of[Ca²⁺]_i was increased by using Ang??,but which was lower significantly by using DHBF and Ang??(P<0.05). The enzymatic reaction of fragmentation cell results showed the activity of Ca²⁺-ATP was decreased by using Ang??, but which was increased significantly by using DHBF and Ang??(P<0.05). Colorimetry results showed the concentration of NO and the activity of NOS were decreased by using Ang??, but which was increased significantly by using DHBF and Ang??(P<0.05). CONCLUSION DHBF can improve the activity of hypertrophy cardiomyocytes which were induced by ang??, downregulate expression of mRNA of ANP and BNP, reduce surface area of cardiomyocytes induced by Ang??,the mechanism of action may be related to promoting the release of NO, regulating the concentration of[Ca²⁺]_i and the activity of Ca²⁺-ATP.     

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??OBJECTIVE To explore the effects of luteolin on the proliferation and migration of the endothelial cells co-cultured with cancer cells and its molecular mechanism. METHODS Human umbilical vein endothelial cells (HUVECs) were primarily isolated and identified by the expression of VE-cadherin. Cancer-endothelial cell co-culture model was established using the Transwell system. The effects of luteolin at different concentrations (0 [Co-culture control], 20 and 50 ??mol??L^-1) on the proliferation and migration of HUVECs in the co-culture system were determined. A HUVECs control group removed of prostate cancer PC3 cells was also included. Human angiogenesis antibody array kit was used to assay the secretion levels of various protein factors in each group. RESULTS VE-cadherin was expressed on all the cultured HUVECs. Increased proliferation ability was found in the HUVECs co-cultured with PC3 cells compared with that in HUVECs control group (P<0.01). Treatment with 20 or 50 ??mol??L^-1 luteolin significantly inhibited the proliferation ability of the HUVECs in co-culture system (both P<0.01). Increased migration ability was found in the HUVECs co-cultured with PC3 cells compared with that in HUVECs control group (P<0.01). Treatment with 20 or 50 ??mol??L^-1 luteolin significantly inhibited the migration ability of the HUVECs in co-culture system (both P<0.01). Secretion levels of multiple angiogenesis-related proteins in the cultural supernatant of co-culture system were significantly increased compared with those in HUVECs control group. Treatment with 20 ??mol??L^-1 luteolin significantly inhibited the secretion levels of interleukin-8 (IL-8), vascular endothelial growth factor (VEGF) and monocyte chemoattractant protein-1(MCP-1) in the cultural supernatant of co-culture system (all P<0.01). CONCLUSION Luteolin may inhibit the proliferation and migration of endothelial cells via suppressing the secretions of IL-8, VEGF and MCP-1 in cancer-endothelial co-culture system.