• Title/Summary/Keyword: pH Inhibition

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Autophagic Degradation of Caspase-8 Protects U87MG Cells Against H2O2-induced Oxidative Stress

  • Zhang, Yi-Bo;Zhao, Wei;Zeng, Rui-Xia
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.7
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    • pp.4095-4099
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    • 2013
  • Oxidative stress induces apoptosis in many cellular systems including glioblastoma cells, with caspase-8 activation was regarded as a major contribution to $H_2O_2$-induced cell death. This study focused on the role of the autophagic protein p62 in $H_2O_2$-induced apoptosis in U87MG cells. Oxidative stress was applied with $H_2O_2$, and cell apoptosis and viability were measured with use of caspase inhibitors or autophagic mediators or siRNA p62, GFP-p62 and GFP-p62-UBA (del) transfection. We found that $H_2O_2$-induced U87MG cell death was correlated with caspase-8. To understand the role of p62 in MG132-induced cell death, the levels of p62/SQSTM1 or autophagy in U87MG cells were modulated with biochemical or genetic methods. The results showed that the over-expression of wild type p62/SQSTM1 significantly reduced $H_2O_2$ induced cell death, but knockdown of p62 aggravated the process. In addition, inhibition of autophagy promoted p62 and active caspase-8 increasing $H_2O_2$-induced apoptosis while induction of autophagy manifested the opposite effect. We further demonstrated that the function of p62/SQSTM1 required its C-terminus UBA domain to attenuate $H_2O_2$ cytotoxity by inhibition of caspase-8 activity. Our results indicated that p62/SQSTM1 was a potential contributor to mediate caspase-8 activation by autophagy in oxidative stress process.

Growth Inhibition of E. coli O157:H7 and Salmonella typhimurium by Lactic Acid Bacteria and Bifidobacteria (젖산균과 비피더스균에 의한 Escherichia coli O157:H7과 Salmonella typhimurium의 생장억제)

  • 김현욱;안영태;신필기
    • Journal of Food Hygiene and Safety
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    • v.12 no.3
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    • pp.181-187
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    • 1997
  • Lactobacillus acidophilus NCFM, Lactobacillus casei YIT 9018, Bifidobacterium longum 8001, and Bifidobacterium longum 8025 at the level of 106 cfu/$m\ell$ were cultured with 104 cfu/$m\ell$ of Escherichia coli O157:H7 KSC 109 or Salmonella typhimurium ATCC14028, in order to verify the effects of lactic acid bacteria and bifidobacteria on the growth of the pathogens. In the mixed culture of lactic acid bacteria with E. coli O157:H7 KSC 109, Growth inhibition and atypical microcolonies of E. coli O157:H7 KSC 109 were observed. The pathogens inoculated grew for 5 hors (pH 5.3), by the time L. acidophilus NCFM reached the exponential growth phase, and then the surviving pathogens were decreased to 101 cfu/$m\ell$ after 35 hours. When L. caseiYIT 9018 was grown with the pathogens, they grew for 10 hours (pH 4.6), by the time L. casei YIT 9018 reached the end of exponential growth phase, and then the surviving pathogens were decreased drastically. Up to the stationary growth phase of lactic acid bacteria, L. acidophilus NCFM exhibited stronger inhibition against the pathogens than L. casei YIT 9018 did, which might be attributed to its faster growth. Likewise bifidobacteria inhibited the growth of the pathogens tested, bifidobaceria was weaker in the inhibitory activity than lactic acid bacteria. When Bifidobacterium longum 8001 was cultured with the pathogens, E. coli O157:H7 KSC 109 was gradually ingibited at the stationary growth phase of bifidobacteria, atypical microcolonies were formed on Levine EMB medium after 48 hours, and Salmonella grew up to 106 dfu/$m\ell$, then was drastically ingibited at the exponential growth phage of Bifidobacterium longum 8001. But when Bifidobacteriuam longum 8025 was cultured with the pathogens, the pathogens grew to the same level of Bifidobacteriuam longum 8025 was cultured with the pathogens, the pathogens grew to the same lever of Bifidobacteriuam longum 8025 after 10 hours, then the surviving pathogens were decreased drastically.

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Characterization of Aspartate Aminotransferase Isoenzymes from Leaves of Lupinus albus L. cv Estoril

  • Martins, Maria Luisa Louro;De Freitas Barbosa, Miguel Pedro;De Varennes E Mendonca, Amarilis Paula Alberti
    • BMB Reports
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    • v.35 no.2
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    • pp.220-227
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    • 2002
  • Two aspartate aminoransferase (EC 2.6.1.1) isoenzymes (AAT-1 and AAT-2) from Lupinus albus L. cv Estoril were separated, purified, and characterized. The molecular weight, pI value, optimum pH, optimum temperature, and thermodynamic parameters for thermal inactivation of both isoenzymes were obtained. Studies of the kinetic mechanism, and the kinetics of product inhibition and high substrate concentration inhibition, were performed. The effect of some divalent ions and irreversible inhibitors on both AAT isoenzymes was also studied. Native PAGE showed a higher molecular weight for AAT-2 compared with AAT-1. AAT-1 appears to be more anionic than AAT-2, which was suggested by the anion exchange chromatography. SDS-PAGE showed a similar sub-unit molecular weight for both isoenzymes. The optimum pH (between 8,0 and 9.0) and temperature ($60-65^{\circ}C$) were similar for both isoenzymes. In the temperature range of $45-65^{\circ}C$, AAT-2 has higher thermostability than AAT-1. Both isoenzymes showed a high affinity for keto-acid substrates, as well as a higher affinity to aspartate than glutamate. Manganese ions induced an increase in both AAT isoenzymes activities, but no cooperative effect was detected. Among the inhibitors tested, hydroxylamine affected both isoenzymes activity by an irreversible inhibition mechanism.

Biological Activities of a Korean Traditional Prescription, Nogyongdaebotang (전통 한약탕제인 녹용대보탕의 생리활성 효과)

  • 이경애;정혜영
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.33 no.1
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    • pp.28-33
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    • 2004
  • This study was performed to examine in vitro biological activities such as antioxidative, nitrite scavenging effect, tyrosinase inhibitory effect and antithrombotic activity of a Korean traditional prescription, Nogyong-daebotang, composed of oriental medical herbs and antler, nourishing the blood, helthiness, strengthening of vital power and promotion of growth. The concentration of total phenolic compounds of the prescription sample was 151.3$\pm$2.6 mg%. The electron donating abilities (EDA) by reduction of 2.2'-diphenyl-1-picrylhydrazyl (DPPH) was 80.9%, inhibition rate of lipid peroxidation in thiocyanate method used linoleic acid was 88.1%. Nitrite scavenging effects of the sample were more than 70% at acidic pH, but it was pH dependent, showing the highest at pH 1.2 and the lowest at pH 6.0. Inhibition rate against tyrosinase of the sample was above 80%. Platelet aggregation induced by ADP (adenosine-5'-diphosphate) was inhibited up to 30% and the inhibitory effect was dependent on concentration of the sample.

Tumor Inhibition Effects and Mechanisms of Angelica sinensis and Sophorae flavescentis ait Decoction Combined with Cisplatin in Xenograft Mice

  • Yan, De-Qi;Liu, Yong-Qi;Li, Ying-Dong;Li, Dou;Cheng, Xiao-Li;Wu, Zhi-Wei
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.11
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    • pp.4609-4615
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    • 2014
  • Background: To investigate tumor inhibition effects and mechanisms of Angelica sinensis and Sophorae flavescentis ait decoction (ASSF) combined with diamine-dichloroplatinum (DDP). Materials and Methods: Bodyweight, tumor inhibition rate and q value were calculated for single ASSF or ASSF combined with DDP on H22 carcinoma xenograft KM mice. Biochemical methods for serum LDH, AST, ALT, and AKP, ELISA method for serum HIF-$1{\alpha}$, pathological assessemnt of thymus, immunohistochemistry detection of tumor tissue caspase3 and mutant p53 protein, and qRT-PCR detection of bax/ bcl-2 mRNA were applied. Results: Compared with DDP control group, the bodyweight increased in ASSF-DDP group (p<0.01). Tumor inhibition rates for DDP, ASSF, ASSF-DDP were 62.7%. 43.7% and 71.0% respectively, with a q value of 0.90. Compared with other groups, thymus of DDP control group had obvious pathological injury (p<0.01), serum LDH, AST, ALT, AKP increased significantly in DDP control group (p<0.01), while serum HIF-$1{\alpha}$ was increased in the model control group. Compared with this latter, the expression of mutant p53 protein and bcl-2 mRNA were decreased in all treatment groups (p<0.01), but there were no statistical difference between DDP control p and ASSF-DDP groups. The expression of caspase3 protein and bax mRNA was increased in all treatment groups, with statistical differences between the DDP and ASSF-DDP groups (p<0.01). Conclusions: ASSF can inhibit bodyweight decrease caused by DDP, can inhibit tumor growth synergistically with DDP mainly through increasing serum HIF-$1{\alpha}$ and pro-apoptotic molecules such as caspase 3 and bax, rather than through decreasing anti-apoptotic mutant p53 and bcl-2. ASSF can reduce DDP toxicity due to decreasing the release of LDH, AST, ALT, AKP into blood and enhancing thymus protection.

Onion peel extract and its constituent, quercetin inhibits human Slo3 in a pH and calcium dependent manner

  • Wijerathne, Tharaka Darshana;Kim, Ji Hyun;Kim, Min Ji;Kim, Chul Young;Chae, Mee Ree;Lee, Sung Won;Lee, Kyu Pil
    • The Korean Journal of Physiology and Pharmacology
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    • v.23 no.5
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    • pp.381-392
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    • 2019
  • Sperm function and male fertility are closely related to pH dependent $K^+$ current (KSper) in human sperm, which is most likely composed of Slo3 and its auxiliary subunit leucine-rich repeat-containing protein 52 (LRRC52). Onion peel extract (OPE) and its major active ingredient quercetin are widely used as fertility enhancers; however, the effect of OPE and quercetin on Slo3 has not been elucidated. The purpose of this study is to investigate the effect of quercetin on human Slo3 channels. Human Slo3 and LRRC52 were co-transfected into HEK293 cells and pharmacological properties were studied with the whole cell patch clamp technique. We successfully expressed and measured pH sensitive and calcium insensitive Slo3 currents in HEK293 cells. We found that OPE and its key ingredient quercetin inhibit Slo3 currents. Inhibition by quercetin is dose dependent and this degree of inhibition decreases with elevating internal alkalization and internal free calcium concentrations. Functional moieties in the quercetin polyphenolic ring govern the degree of inhibition of Slo3 by quercetin, and the composition of such functional moieties are sensitive to the pH of the medium. These results suggest that quercetin inhibits Slo3 in a pH and calcium dependent manner. Therefore, we surmise that quercetin induced depolarization in spermatozoa may enhance the voltage gated proton channel (Hv1), and activate non-selective cation channels of sperm (CatSper) dependent calcium influx to trigger sperm capacitation and acrosome reaction.

Removal and Regrowth Inhibition of Microcystis aeruginosa using Artemisia asiatica Extracts (쑥 추출액을 이용한 Microcystis aeruginosa 제거 및 성장억제 연구)

  • Choi, Hee-Jeong
    • Journal of Korean Society on Water Environment
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    • v.33 no.4
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    • pp.441-448
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    • 2017
  • Microcystis aeruginosa (M. aeruginosa) is a cyanobacterium species that can form harmful algal blooms in freshwater bodies worldwide. The use of Artemisia asiatica extracts to control M. aeruginosa inhibition will be environmentally friendly and promising. Artemisia asiatica extracts removed successfully upto 88% of M. aeruginosa pH 8 at $25^{\circ}C$ of temperature. These results was indicated that the amount of 2.24 g/L Artemisia asiatica extracts was removed 1g dryweight/L of M. aeruginosa. The kinetic data showed substrate inhibition kinetics and maximum growth rate was obtained when the M. aeruginosa was grown in medium containing 2.5 g/L of initial concentration of Artemisia asiatica extracts. In the various growth control models, Luong model showed the highest correlation coefficient of 0.9916. Therefore, the Luong model was the most suitable control model for the growth control of M. aruginosa using Artemisia asiatica extracts. In conclusion, the growth control of M. aruginosa using Artemisia asiatica extracts can be applied in the field without controlling the temperature and pH of rivers and streams, and it is possible to control the growth of M. aruginosa efficiently in a short time. The natural extract, Artemisia asiatica extracts, can be a promising inhibition due to its high efficiency and low dose requirements.

The Biological Activity of Deer Antler Extract in vitro (In vitro에 의한 녹용 추출물의 생리 활성 효과)

  • Lee, Kyung-Ae;Chung, Hae-Young
    • The Korean Journal of Food And Nutrition
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    • v.20 no.2
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    • pp.114-119
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    • 2007
  • Our research objective was to examine the in vitro biological activity of deer antler(Nogyong in Korean) extract, including the antioxidative, nitrite scavenging, and tyrosinase inhibitory effects, as well as the antithrombotic, and angiotensin I converting enzyme(ACE) inhibitory activities. The carbohydrate, protein, fat, and mineral contents of the deer antler were 7.6%, 65.3%, 3.2% and 23.9%, respectively. The electron donating ability(EDA) by the reduction of 2,2'-diphenyl-1-picrylhydrazyl(DPPH) was 67.1%, and the inhibition rate of lipid peroxidation by the thiocyanate method using linoleic acid was 92.1% in 100 mg/ml of extract. The nitrite scavenging effects were pH dependent, and were highest at pH 1.2 and lowest at pH 6.0. The sample inhibition rate against tyrosinase was above 64.0%. The platelet aggregation induced by ADP(adenosine-5'diphosphate) was inhibited up to 51.7%, and the inhibitory effect was dependent on the sample concentration. Lastly, the inhibition rate of ACE was 47.5% in 100 mg/ml of deer antler extract.

동물의 장에서 분리한 Enterococcus sp.의 특성 및 분말화

  • Park, Chong-Jin;Pyeon, Jeong-Soo;Cho, Yun-Kyung;Hong, Seung-Suh;Lee, Hyun-Soo
    • Microbiology and Biotechnology Letters
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    • v.24 no.4
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    • pp.393-398
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    • 1996
  • In order to develop a lactic acid bacterial powder which can be used as a probiotic for human and animal, a lactic acid bacteria which has high resistance against low pH and ox-gall, and shows a good growth inhibition against E. coli, was isolated from an animal intestine and characterized. The isolated strain was identified as Enterococcus faecium. It had more than 90% of survival at low pH for 2 hours and almost 100% of survival in the presence of 0.3% ox-gall. When co-cultured with E. coli in MRS broth, all of the E. coli cells were killed within 24 hours. The final powdered product of the isolated strain was manufactured after a freeze drying process using an industrial media, and then checked its stability. Its storage stability was 80% for 11 months at 18$\circ$C.

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Testing of Cancer Chemopreventive Potential of Prunella vulgaris L. Aqua-acupuncture Solution Using Biochemical Markers of Carcinogenesis (발암과정 생화학적 표식자를 이용한 하고초 약침액의 암예방 활성 측정)

  • Park, Sin-Hwa;Cho, Kyoung-Hee;Shon, Yun-Hee;Lim, Jong-Kook;Nam, Kyung-Soo
    • Korean Journal of Pharmacognosy
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    • v.32 no.2 s.125
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    • pp.163-167
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    • 2001
  • Prunella vulgaris L. aqua-acupuncture solution (PVAS) was tested for cancer chemopreventive activity using chemoprevention-associated biochemical end points. The following effects were measured. : (a) inhibition of 7,12-dimethylbenz[a]anthracene (DMBA)-induced cytochrome P4501A1 activity (b) inhibition of $[^3H]B[a]P-DNA$ binding (c) inhibition of phorbol 12-myristate 13-acetate (TPA)-induced free radical formation in HL-60 cells (d) inhibition of polyamine metabolism. PVAS inhibited cytochrome P4501A1-mediated ethoxyresorufin-O-deethylase activity. The binding of $[^3H]B[a]P$ metabolites to DNA of NCTC-clone 1469 cells was inhibited significantly by PVAS. There is 22% inhibition of TPA-induced free radical formation in human leukemic cells with 5 mg/ml PVAS. Proliferation of Acanthamoeba castellanii was inhibited by PAVS at concentration of 30 mg/ml. PAVS positive in these assays may inhibit the carcinogenesis process and is considered very promising cancer-preventing agent because of its multiple activities.

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