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Objective:To detect the repair effect of brain protein hydrolyzate(BPH)on the oxidative damage induced by H2O2in the PC12 cells,and to optimize the prescription of BPH stomach floating tablets using the star design effect surface method.Methods:The PC12 cells in the logarithmic phase were divided into normal control group,model group(300 μmol·L-1H2O2),BPH group,artificial gastric juice-treated BPH(GBPH)group, artificial intestinal juice-treated BPH(IBPH)group,artificial gastric juice-treated BPH tablets(GBPH-T)group and artificial gastric juice-treated BPH floating tablets(GBPH-FT)group(The latter five groups were added with 20,40,60,80 and 100 mg·L-1BPH treated with different conditions);at the same time blank control group was set up.The PC12 cells in normal control group didn't receive any treatment,the blank control group was added with medium only,and the PC12 cells in model group were treated with H2O2(300 μmol·L-1)for 3 h. The cell viability was detected by MTT assay.Using Design-expert 8.0.6 Trial software,the dosages of HPMC-K4M,octadecanol and acrylic resin Ⅱ were used as the investigation factors,and the 8 h cumulative release in vitro was used as the evaluation index to optimize the prescription.Results:Compared with normal control group, the activity of PCl2cells in 60 mg·L-1BPH group was increased(P<0.05).Compared with model group,the vitalities of PC12 cells in BPH group,IBPH group,GBPH group and GBPH-FT group were increased significantly (P<0.05 or P<0.01).Compared with BPH group,the cell viability in IBPH group was decreased significantly (P<0.05),and there was no significant difference in GBPH group(P>0.05).Compared with GBPH-T group, the cell viability in GBPH-FT group was significantly increased(P<0.05).The optimal prescription was BPH 20 mg,lactose 10 mg,magnesium stearate 0.4 mg,microcrystalline cellulose 20 mg,HPMC-K4M 27 mg, octadecanol 63 mg,acrylic resinⅡ13 mg;all floating tablets drift in 5 s and sustained floating > 8 h;the difference of the measured value of 8 h cumulative release and the predicted value was not statistically significant (P>0.05).Conclusion:BPH has a significant repair effect on the H2O2-induced oxidative damage in the PC12 cells.Star design-effect surface method has good predictability and reproducibility;it is reasonable and feasible, and can be used to optimize the prescription of BPH stomach floating tablets.
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Objective:To observe the protective effect of deproteinized extract of calf blood (DECB) on the kidney injury of the diabetic rats, and to discuss its mechanism preliminarily.Methods:The male Wistar rats were intraperitoneally injected with STZ (65 mg·kg-1) to establish the diabetes models, then the model rats were randomly divided into model (M)group, and metformin (MMet, 105 mg·kg-1) group , low dose of combined administration (ML,105 mg·k-1 metformin+94.5 mg·kg-1 ,DECB) group, medium dose of combined administration(MM, 105 mg·kg-1 metformin+ 189 mg·kg-1 DECB) group, high dose of combined administration(MH, 105 mg·kg-1 metformin +378 mg·kg-1,DECB) group, another ten Wistar rats were selected as normal control(NC)group.The rats were intragastrically administed with metformin and intraperitoneally injected with DECB, once a day, total of 8 weeks.The rats in NC group and M group were given normal saline solution.The weights and blood glucose levels of the rats in various groups were determined;the levels of serum high density lipoprotein-cholesterol (HDL-C), low density lipoprotein-cholesterol (LDL-C), blood urea nitrogen (BUN), urinary albumin (UAlb), urine creatinine (UCR), serum creatinine (SCr), total cholesterol (TC), triglyceride (TG), uric acid (UA), glutathione (GSH), and malondialdehyde (MDA) were detected;the activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) were determined.The pathological changes of kidney tissue of the rats in various groups were detected by HE staining.Results:Compared with NC group, the weight of the rats in M group was reduced(P<0.05),and the levels of blood glucose,UAlb,UCr,SCr,UA,BUN,LDL-C,TC,TG,and MDA were increased(P<0.05);the levels of HDL-C and GSH were obviously reduced(P<0.05),and the activities of SOD adn GSH-Px were obviously reduced(P<0.05).Compared with M group,the weights of the rats in MM and MH groups were increased (P<0.05), and the levels of blood glucose,UAlb,UCr,SCr,UA,BUN,LDL-C,TC,TG, and MDA were decreased (P< 0.05);the levels of HDL-C and GSH were increased (P<0.05),and the activities of SOD and GSH-Px were increased (P<0.05).The pathological observation of kidney tissue showed that the rats in M group had obvios kidney tissue lesions with glomerular congestion and renal tubular edema compared with NC group;compared with M group,the pathological changes of the kidney tissue of the rats in drug administration groups were significantly improved, the renal tubular vacuoles were reduced, and the interstitial hyperplasia was not obvious.Conclusion:DECB combined with metformin can reduce the blood glucose level, regulate blood lipid, improve the pathological changes of kidney tissue in the diabetic rats, reduce the renal damage, and enhance the antioxidant capacity of the body.
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Objective:To observe the protective effect of deproteinized extract of calf blood (DECB)on the ethanol-induced liver injury of the mice,and to preliminaryly discuss its mechanism. Methods:Sixty healthy ICR mice were divided into control group,model group,positive drug group,low,medium and high doses of DECB groups (n=10).By intragastric administration,the mice in control group were given 20 mL·kg-1 saline solution, the mice in low,medium and high doses of DECB groups were administrated with 0.125,0.250,0.500 g·kg -1 DECB,and the mice in positive drug group were administrated with 0.63 g·kg -1 Hugan Tablets;once a day for 30 d. 1 h after the last administration,except control group,the mice in other groups were administrated with one-time grant of 50% ethanol 14 mL·kg -1 ,and fasted for 16 h to establish the models of acute alcohol liver injury.The endurance alcohol time and drunk time of the mice were determined,the activities of aspartate aminotransferase (ALT)and alanine transaminase (AST)activity in serum of the mice were detected,the levels of triglyceride (TG),glutathione (GSH)and malonic dialdehyde (MDA)in liver tissue were determined,and the pathological changes of liver tissue were detected.Results:Compared with model group,the drunk symptoms of the mice in different doses of DECB groups were obviously reduced,the endurance time of the mice in high dose of DECB group and positive drug group was prolonged (P <0.05),and the drinking time was shortened (P <0.05);the ALT and AST activities in serum in mediun and high doses of DECB groups were significantly lower than those in model group (P <0.05).Compared with model group,the MDA and TG levels in liver tissue of the mice in medium and high doses of DECB groups and positive drug group were obviously reduced,and the GSH levels were increased (P <0.05);compared with model group,the pathological damages of liver tissue of the mice in high dose of DECB group caused by ethanol were significantly reduced.Conclusion:DECB can improve ethanol-induced liver injury which may be related to the inhibition of hepatic oxidative stress response.
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@#Objective To investigate the effect of thiopental on lipopolysaccharide(LPS)-induced activation of nuclear factor kappa B(NF-κB) under different concentration or after different time in rat glioma cells in vitro.Methods The rat C 6 glioma cells were treated with,in the presence or Absence of various concentrations(10-3 mol/L,10-4 mol/L and 10-5 mol/L)of thiopental for 1 h.30 min before harvest,the cells were exposed to LPS(10 ng/ml).The other C 6 cells were treated with 10-5 mol/L thiopental with different time(0.5 h,1 h,2 h,4 h).30 min before harvest,the cells were exposed to LPS(10 ng/ml).Nuclear extracts from C 6 cells were harvested using standard procedures,and the LPS-induced NF-κB expression was determined with electrophoretic mobility shift assays(EMSA) and measured with densitometry(Bio-Rad).Results The width of the bands of NF-κB under thiopental with different concentrations were similar,but they were narrower than that of the control(treated only with LPS),the density was less than that of the control(P<0.05).The width of the bands of NF-κB under thiopental with different time decreased with the time(P<0.05),and they were much narrower and shallower than that of the control(P<0.05),and the density was significantly less than that of the control(P<0.05).Conclusion Thiopental suppressed the activation of transcription factor NF-κB,and the inhibition was concentration-independent but time-dependent.
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Objective To study the anti-ageing and antioxidation of polyphends of vitis amurensis Rupr(PVAR).Methods Forty senile male Wistar rats (20 months) were divided into 4 goups according rats′ weights:senile negtive control group(A), fed with basic forage;two senile PVAR groups(B and C),fed with basic forage inchuding 200 and 400 mg?kg~ -1 ?d~ -1 PVAR;senile positive control(D), fed with basic forage inchuding 200 mg?kg~ -1 ?d~ -1 melissic powder.After fed continuously for 8 weeks,the MDA contents in RBC-membrane, plasma and liver,sulfhydryl content in RBC-membrane, sialic acid content,activity of Na~+-K~+-ATPasc, fluidity of RBC-membrane,and activity of plasma GSH-Px in rats were detected.Results Compared with group A,the contents of MDA in groups B,C,and D decreased significantly(P
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angiotensi I.Conclusion Angiotensin Ⅲ is a good natural substrate for TPP Ⅰ.
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OBJECTIVE: To study the anti-lipid peroxidation action of Rosa davurica Pall. fruit. METHODS: The contents of malondialdehyde (MDA) of the tissue homogenate of the liver, heart, kidney, brain and of the red blood cells induced by hydrogen peroxide in mice were measured. The contents of MDA and the activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) of ischemic myocardium of mice were measured. RESULTS: 0.2 g/L Rosa davurica Pall. fruit could decrease significantly the contents of MDA of the all tissue (P < 0.05). Inhibition rate of 6.7 g/L Rosa davurica Pall. fruit on MDA of the red blood cells induced by hydrogen peroxide was 89.2%. Administration of this extraction successively for six days (ig, 2.0 g x kg(-1) x d(-1)) can significantly reduce the content of MDA (P < 0.01) and augment the activities of SOD and GHS-Px (P < 0.05) in the ischemic myocardium of mice. CONCLUSION: Rosa davurica Pall. fruit can significantly prevent the lipid peroxidation.
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Objective:To analyze OLFM1 by bioinformatics and prepare its polyclonal antibody.Methods:According to the bioinformatics analysis and prediction of the possible high structure,hydrophobicity and antigenicity of OLFM1 and the principal of antibody project,a partial peptide of OLFM1 with 20-amino acid residues with was synthesized after homology search.The synthesized peptide was then used to immunize rabbit.The specificity and titer of polyclonal antibody against OLFM1 were identified via ELISA and Western blot.Results:Polyclonal antibody against OLFM1 was proved to recognize OLFM1 protein specifically,and its titers reached 1?32 000.Conclusion:By the bioinformatics analysis and prediction,the hydrophilicity and antigenicity of OLFM1 are analyzed.The polyclonal antibody against OLFM1 is successfully obtained.
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Objective:To prepare gACE polyclonal antibody for functional study of gACE.Methods:According to the bioinformatics analysis and prediction of the possible conformational structure,hydrophobicity and antigenicity of gACE and the principal for antibody production,a partial peptide with 18-amino acid residues of gACE was synthesized after homology search.The synthesized peptide was then used to immunize after coupling with KLH.The properties of anti-gACE were analyzed by ELISA,Western blot and immunohistochemistry.Results:The antigenicity was repredicted by bioinformatics analysis.The polyclonal antibody against gACE was successfully obtained and its specificity and sensitivity we conformed by ELISA,Western blot and immunohistochemistry.Conclusion:By the bioinformatics analysis and prediction,the hydrophilicity and antigenicity of gACE are analyzed.The antibody of gACE is successfully obtained.