ABSTRACT
Objective:To investigate the effect of Stigma Maydis Palysaccharide(SMPS)on ATP synthesis in kidney mitochondria of D-galactose-induced aging mice, and to clarify its possible mechanism.Methods:The aging mouse model was established by subcutaneous injection of D-galactose solution in the back of the neck.The 48 SPF male mice were randomly divided into normal control group(control group), D-galactose model group(D-Gal group), SMPS low-dose group and SMPS high-dose group(n=12 for each). The control group was subcutaneously injected with 150 mg/kg normal saline on the back of the neck every day, while the other three groups were subcutaneously injected with 150 mg/kg of D-gal solution on the back of the neck every day.SMPS-L and-H dose groups were given 30 mg/kg and 60 mg/kg of SMPS solution by gavage at the same day of D-Gal injection.The control group and D-GAL group were given the same volume of normal saline daily by gavage for 42 days.Blood samples were collected from the eyeball under general anesthesia after 42 days of intervention for the detection of serum levels of superoxide dismutase(SOD), glutathione peroxidase(GSH-Px)and MDA.After harvesting the kidney tissue, various tests were used to detect ATP content, the mRNA expression levels and protein expression levels in kidney.Luciferase assay was used to detect ATP content in renal tissue.Real-time fluorescent quantitative PCR was used to detect the mRNA expression levels of succinate dehydrogenase(SDH)of complex Ⅱ, cytochrome C reductase(Cycs)of complex Ⅲ, complex Ⅳ(COXⅣ)and ATP5b in ATP synthase in mitochondrial oxidative respiratory chain.Western blot was used to detect the expression levels of mitochondrial fusion protein 2(MFN2), dynamin-related protein1(DRP1)and mitochondrial autophagy related protein P62 in renal tissues of each group.Results:Compared with control group, the activities of serum of SOD(116.53±10.01)U/mg and GSH-Px(127.58±8.74)μmol/L were significantly decreased in D-GAL group(both P< 0.01), and serum MDA content(15.42±0.91)μmol/L increased significantly in D-GAL group( P<0.01). Compared with D-GAL group, the activities of SOD(152.80±9.29)U/mg and GSH-Px(274.07±10.73)μmol/L were significantly increased in SMPS intervention group( P< 0.01), while the MDA content(8.10±0.66)μmol/L decreased significantly in SMPS intervention group( P< 0.01). Compared with control group, the content of ATP(178±4)10 -4 μmol in D-gal group was significantly decreased( P<0.01), the mRNA expression levels of SDH, Cycs and COXⅣ were not significantly changed in D-gal group, and the mRNA expression level of ATP5b(0.67±0.01)was down-regulated in D-gal group( P<0.01), the expression of MFN2 protein(0.29±0.02)was significantly decreased in D-gal group( P<0.01), and the expression of DRP1 and P62 protein(0.31±0.02 and 0.21±0.01)was significantly increased in D-gal group(both P<0.01). Compared with the D-gal group, the ATP content(193±1)10 -4 μmol in the kidney tissue of the mice was significantly increased in SMPS intervention group( P< 0.01), and the ATP5b mRNA expression and MFN2 protein expression(0.87±0.05 and 0.71±0.08)were significantly increased in SMPS intervention group(both P< 0.01), DRP1 and P62 protein expressions(0.20±0.01 and 0.10±0.01)were significantly down-regulated in in SMPS intervention group(both P< 0.01). Conclusions:SMPS can improve the mitochondrial dynamic homeostasis disorder in aging mice by increasing the activity of antioxidant enzymes, up-regulating the expression of ATP5b mRNA and MFN2 protein, down-regulating the expression of DRP1 and P62 protein, and promoting the generation of mitochondrial ATP in D-gal-induced aging mice kidney tissue.
ABSTRACT
Objective:To explore the influence of fermented red ginseng extract (FRGE) in the proliferation of rat glomerular mesangial cells (GMCs) and the degradation of extracellular matrix(ECM)under high sugar stimulation, and to clarify the prevention and treatment effects of FRGE on diabetic nephropathy (DN) and the possible mechanism.Methods:The rat GMCs were cultured and divided into normal concentration of D-glucose (NG) group, high concentration of D-glucose (HG) group and high concentration of D-glucose plus different concentrations (3.75, 7.50, 15.00 mg·L-1) of FRGE groups. The proliferation rates of rat GMCs were detected with MTT,and the type Ⅳcollagen(Col Ⅳ) levels in supernatants of the GMCs were detected by ELISA. The protein expressions levels of matrix metalloproteinase-2(MMP-2) and tissue inhibitor of metalloproteinase-2 (TIMP-2) were detected with Western blotting m ethod.Results:Compared with NG group, the proliferation rate of GMCs in HG group was increased(P<0.01), the Col Ⅳ level was increased(P<0.01),the MMP-2 expression level was decreased, and the TIMP-2 expression level was up-regulated(P<0.01).Compared with HG group, the proliferation rates of GMCs in various FRGE groups were decreased(P<0.01), the Col Ⅳ levels were decreased(P<0.01),the expression levels of TIMP-2 were reduced(P<0.01),and the expression levels of MMP-2 were increased(P<0.01).Conclusion:FRGE can inhibit the proliferation of rat GMCs induced by high sugar and promote the ECM degradation to delay the occurrence and development of DN.
ABSTRACT
Objective: To explore the inhibitory effect of gallotannin (GLTN) on the proliferation of rat glomerular mesangial cells (GMC)induced by high sugar stimulation and the influence in the cell cycle of the rats, and to clarify the prevention effect of GLTN in the occurrence and development of diabetic nephropathy (DN). Methods:The experimental cells were divided into normal control group (D-glucose 5.5 mmol·L-1 ,NC group), high glucose group (D-glucose 30 mmol· L-1 ,HC group),high glucose + 5 mmol· L-1 3 - AB group (AB group),high glucose + 20 μmol·L-1 GLTN group (G20 group),high glucose + 40 μmol· L-1 GLTN group (G40 group).The proliferation of GMC in different groups at different time points (4,8,24,48 and 72 h)was observed by MTT assay.The changes of cell cycle of GCM under different culture conditions were examined by flow cytometry,and the expression levels of TGF-β1 and CTGF were detected by Western blotting method. Results:Compared with NC group,the proliferation levels of GMC in HC group were increased (P <0.01),and reached the peak at 48 h ;the percentage of S phase cells was increased (P <0.01).Compared with HC group,the proliferation levels of GMC in 3-AB group and GLTN group were significantly decreased (P < 0.01 ),and the percentages of S phase cells were decreased (P <0.01).Compared with NC group,the protein expression levels of TGF-β1 and CTGF in each drug group were increased (P < 0.05 or P < 0.01),but they were significantly lower than those in HC group (P < 0.01).Conclusion:GLTN can inhibit the proliferation of GMC under high sugar stimulation through arresting the cell cycle and down-regulating the expressions of TGF-β1 and CTGF and delay the occurrence and development of DN.
ABSTRACT
<p><b>OBJECTIVE</b>To establish a convenient and accurate method of DNA molecular marker for the identification of corium stomachium galli.</p><p><b>METHOD</b>Cytb mtDNA sequences of Gallus gallus domestica and three other species of poultry were downloaded from Genbank. Species-specific PCR primers were designed according to the differential DNA fragments of the cytb genes. PCR tests were performed with the DNAs extracted from G. gallus domestica, three other poultry species and domestic mammal animals.</p><p><b>RESULT</b>The specific primers of G. gallus domestica could only amplify the cytb mtDNA of G. gallus domestica.</p><p><b>CONCLUSION</b>The primers are specific to G. gallus domestica mtDNA and can used to discern Corium stomachium from the false medicine.</p>
Subject(s)
Animals , Chickens , Classification , Genetics , DNA, Mitochondrial , Genetics , Liver , Metabolism , Polymerase Chain ReactionABSTRACT
Objective To observe the structure changes of the synapse of the neurons in hippocampus CA3 of young rats and study the basis for the mechanism of polyene phosphatidyl in providing learning and memory and the effect on synaptic plasticity.Methods A total of 20 Wistar rats with 5 months were randomly divided into polyene phosphatidyl group and normal control group.Each group had 10 rats.After 4 weeks feeding,Water maze training was peformed in all the rats for 1 weeks.The immue expressions of synapsin(SYN) of the rats in polyene phosphatidyl group and control groups were observed with immunohistochemical method and analyzed by MOTAC imagine analyzing system.The change of synapse of hippocamal CA3 was observed with electron microscope.And the other 24 to 26 months rats were selected as aged group,and fed in the same condition.Moreover,the ultrastructures of hippocamal CA3 of aged rats were observed by transmission electron microscopy(TEM).Results The SYN in polyene phosphatidyl group(0.430 0?0.022 4) was higher than that in control group(0.3567?0.0209) (P
ABSTRACT
Objective:To study the cytotoxicity to tumor cells P815,H22 and B16 in vitro and the treatment effect of mice spleenocytes activated by lymphokine IL-2 and Tremella Polysaccharide,and their effects on main viscerals.Methods:The cytotoxicity of spleenocytes activated by lymphokine IL-2 or IL-2 and Tremella Polysaccharide to P815,H22 and B16 was detected by ?-scintillation counter after 2 hours.Subcutaneous injection of the above-mentioned spleenocytes to the local area of jepatocarcinoma of mice,one injection every two days,5 times totally.To examine tumor weight,size,histological changes and morphological changes in main viscerals.Results:The significantly cytotoxicity to tumor cells of spleenocytes activated by IL-2 and Tremella Polysaccharide was observed.To B16,the cytotoxicity of activated spleenocytes was the strongest.To H22,the cytotoxicity of activated spleenocytes was stronger and to P815,the cytotoxicity was the weakest,the cytotoxicity of TP-LAK cells was significantly stronger than that of LAK cells( P