• 제목/요약/키워드: enzymatic pretreatment

검색결과 181건 처리시간 0.033초

Effects of Histamine on Cultured Interstitial Cells of Cajal in Murine Small Intestine

  • Kim, Byung Joo;Kwon, Young Kyu;Kim, Euiyong;So, Insuk
    • The Korean Journal of Physiology and Pharmacology
    • /
    • 제17권2호
    • /
    • pp.149-156
    • /
    • 2013
  • Interstitial cells of Cajal (ICCs) are the pacemaker cells in the gastrointestinal tract, and histamine is known to regulate neuronal activity, control vascular tone, alter endothelial permeability, and modulate gastric acid secretion. However, the action mechanisms of histamine in mouse small intestinal ICCs have not been previously investigated, and thus, in the present study, we investigated the effects of histamine on mouse small intestinal ICCs, and sought to identify the receptors involved. Enzymatic digestions were used to dissociate ICCs from small intestines, and the whole-cell patch-clamp configuration was used to record potentials (in current clamp mode) from cultured ICCs. Histamine was found to depolarize resting membrane potentials concentration dependently, and whereas 2-PEA (a selective H1 receptor agonist) induced membrane depolarizations, Dimaprit (a selective H2-agonist), R-alpha-methylhistamine (R-alpha-MeHa; a selective H3-agonist), and 4-methylhistamine (4-MH; a selective H4-agonist) did not. Pretreatment with $Ca^{2+}$-free solution or thapsigargin (a $Ca^{2+}$-ATPase inhibitor in endoplasmic reticulum) abolished the generation of pacemaker potentials and suppressed histamine-induced membrane depolarization. Furthermore, treatments with U-73122 (a phospholipase C inhibitor) or 5-fluoro-2-indolyl des-chlorohalopemide (FIPI; a phospholipase D inhibitor) blocked histamine-induced membrane depolarizations in ICCs. On the other hand, KT5720 (a protein kinase A inhibitor) did not block histamine-induced membrane depolarization. These results suggest that histamine modulates pacemaker potentials through H1 receptor-mediated pathways via external $Ca^{2+}$ influx and $Ca^{2+}$ release from internal stores in a PLC and PLD dependent manner.

이진탕의 생쥐 소장 카할세포 향도잡이 전압에 미치는 효능에 관한 연구 (Effects of Yijin-tang on Pacemaker Potentials in Interstitial Cells of Cajal of Murine Small Intestine)

  • 한동훈;김정남;김병주
    • 대한한의학방제학회지
    • /
    • 제28권1호
    • /
    • pp.71-80
    • /
    • 2020
  • Obejectives : The purpose of this study was to investigate the effects of Yijin-tang on pacemaker potentials of small intestinal interstitial Cells of Cajal (ICC). Methods : To dissociate the ICC, we used enzymatic digestions from the small intestine in mice. The electrophysiological whole-cell patch-clamp configuration was used to record pacemaker potentials in the cultured ICC and the in vivo effects of Yijin-tang on GI motility were investigated by calculating percent intestinal transit rates (ITR). Results : 1. The ICC generated pacemaker potentials in the murine small intestine. Yijin-tang produced membrane depolarization with concentration-dependent manners in the current clamp mode. 2. Pretreatment with a Ca2+ free solution and thapsigargin, a Ca2+-ATPase inhibitor in the endoplasmic reticulum, stopped the pacemaker potentials. In the case of Ca2+-free solutions and thapsigargin, Yijin-tang did not induce membrane potential depolarizations. 3. U73122, a phospholipase C (PLC) inhibitors, blocked the Yijin-tang-induced membrane potential depolarizations. However, U73343, an inactive PLC inhibitors, did not block. 4. In the presence of protein kinase C (PKC) inhibitors, staurosporine or Rottlerin, Yijin-tang depolarized the pacemaker potentials. However, in the presence of Go6976, Yijin-tang did not depolarize the pacemaker potentials. 5. In mice, intestinal transit rate (ITR) values were significantly and dose-dependently increased by the intragastric administration of Yijin-tang. Conclusions : These results suggest that Yijin-tang can modulate the pacemaker activity of ICC through an internal/external Ca2+ and PLC/PKC-dependent pathway in ICC. In addition, Yijin-tang is a good candidate for the development of a prokinetic agent.

효소, Trifluoromethanesulfonic Acid, 열, 및 NaOH 처리에 의한 계란 난백의 항원성 변화 (Changes in the Antigenicity of Chicken Egg White by the Treatments of Protease, Trifluoromethanesulfonic Acid, Heat, and NaOH)

  • 류주현;이종미;손동화
    • 한국식품과학회지
    • /
    • 제32권3호
    • /
    • pp.720-725
    • /
    • 2000
  • 여러 가지 처리가 난백의 항원성을 감소시키는데 미치는 영향을 조사하기 위하여, 토끼 항난백항체로 난백 처리물들의 간접 경합 ELISA(ciELISA)를 실시하였다. 식물성, 동물성, 미생물성의 9가지 효소를 이용한 가수분해처리는 난백의 항원성에 영향을 주지 못했다. 보다 효율적인 효소 가수분해를 위해 방사선 조사 후, 효소처리를 실시했는데, 이것 역시 항원성을 감소시키지 못했다. Trifluoromethanesulfonic acid 처리는 난백의 항원성을 거의 제거시키는 것으로 나타났다. 열처리는 $121^{\circ}C$ 고온처리의 경우에서만 반응성이 1/10,000이하로 감소하여 항원성이 거의 사라졌다. NaOH 단독 처리는 0.3%(w/v) 이상의 농도에서 반응성이 감소되기 시작하여, 1%(w/v) 이상에서는 반응성이 1/10,000이하로 감소하였다. NaOH 처리에 $70^{\circ}C$, 15분간의 열처리를 더해 주면, NaOH 단독 처리시보다 효과적으로 항원성이 감소되는 것으로 나타났다.

  • PDF

Protective Effects of Acetylbergenin against Carbon Tetrachloride Induced Hepatotoxicity in Rats

  • Lim, Hwa-Kyung;Kim, Hack-Seung;Kim, Seung-Hwan;Chang, Myung-Jei;Rhee, Gyu-Seek;Choi, Jong-Won
    • Archives of Pharmacal Research
    • /
    • 제24권2호
    • /
    • pp.114-118
    • /
    • 2001
  • The present study was undertaken to investigate whether or not the hepatoprotective activity of acetylbergenin was superior to bergenin in carbon tetrachloride ($CCl_4$)-intoxicated rat. Acetylbergenin was synthesized by acetylating bergenin, which was isolated from Mallotus japonicus. The hepatoprotective effects of acetylbergenin were examined against $CCl_4$-induced liver damage in rats by means of serum and liver biochemical Indices. Acetylbergenin was administered orally once daily for 7 successive days, then a 0.5 ${m/kg}$ mixture of $CCl_4$in olive oil (1:1) was intraperitoneally injected at 12 h and 36 h after the final administration of acetylbergenin. Pretreatment with acetylbergenin reduced the elevated serum enzymatic activities of alanine/aspartate aminotransferase, sorbitol dehydrogenase and $\gamma$-glutamyltransferase in a dose dependent fashion. Acetylbergenin also prevented the elevation of hepatic malondialdehyde formation and depletion of glutathione content dose dependently in $CCl_4$-intoxicates rats. In addition, the decreased activities of glutathione S-transferase and glutathione reductase were restored to almost normal levels. The results of this study strongly suggest that acetylbergenin n has potent hepatoprotective activity against $CCl_4$-induced hepatic damage in rats by glutathione-mediated detoxification as well as having free radical scavenging activity. In addition, acetylbergenin doses of 50 ${mg/kg}$showed almost the same levels of hepatoprotection activity as 100 ${mg/kg}$ of bergenin, indicating that lipophilic acetylbergenin is more active against the antihepatotoxic effects of $CCl_4$ than those of the much less lipophilic bergenin.

  • PDF

Doxorubicin Inhibits the Production of Nitric Oxide by Colorectal Cancer Cells

  • Jung, In-Duk;Lee, Jang-Soon;Yun, Seong-Young;Park, Chang-Gyo;Han, Jeung-Whan;Lee, Hyang-Woo;Lee, Hoi-Young
    • Archives of Pharmacal Research
    • /
    • 제25권5호
    • /
    • pp.691-696
    • /
    • 2002
  • Doxorubicin (DOX) is an active and broad spectrum chemotherapeutic agent. Increased inducible nitric oxide synthase (NOS) expression and/or activity have been reported in several human tumors. While the relationship between DOX treatment and the enzymatic activity of endothelial NOS has been well characterized, little is known about the effects of DOX on the expression of iNOS in human cancer cells. In the present study, we characterized the effects of DOX on the nitric oxide (NO) production by colorectal cancer cells, DLD-1. IFN-${\gamma}$/IL-1$\beta$ (CM) increased the production of NO, whereas pretreatment of DOX inhibited the production of NO in response to CM in a dose dependent manner. The increased expressions of iNOS mRNA and protein by CM were completely blocked by DOX without affecting the iNOS mRNA stability. However, DOX activated nuclear factor-kB (NF-kB) in response to CM. Furthermore, the expression of inhibitor kB$\alpha$ was reduced by DOX in a dose dependent manner. Collectively, DOX inhibited the production of NO by DLD-1 cells, which is not linked to well known transcription factor, NF-kB. Therefore, further studies on the possible mechanisms of inhibitory effects of NO production by DOX would be worth pursuing.

Protective effect of ultrasonication-processed ginseng berry extract on the D-galactosamine/lipopolysaccharide-induced liver injury model in rats

  • Nam, Yoonjin;Bae, Jinhyung;Jeong, Ji Hoon;Ko, Sung Kwon;Sohn, Uy Dong
    • Journal of Ginseng Research
    • /
    • 제42권4호
    • /
    • pp.540-548
    • /
    • 2018
  • Background: Acute hepatic failure is a life-threatening critical condition associated with rapid deterioration of liver function and liver transplantation. Several studies have shown that Panax ginseng Mayer has antidiabetic and hepatoprotective effects. However, the hepatoprotective effect of ginseng berry is still unveiled. In this study, we evaluated the hepatoprotective effects of ultrasonication-processed ginseng berry extract (UGBE) on acute hepatic failure model in rats. Methods: Ginseng berry extract (GBE) was ultrasonically processed. The GBE, silymarin, and UGBE were orally administered to male Sprague-Dawley rats for 4 wk. Twenty-four h after the last administration, rats were challenged with D-galactosamine (D-GalN)/lipopolysaccharide (LPS). Results: After ultrasonication, the component ratio of ginsenosides Rg2, Rg3, Rh1, Rh4, Rk1, Rk3, and F4 in GBE had been elevated. Administration of UGBE significantly increased the survival rate of D-GalN/LPS-challenged rats. Pretreatment with UGBE significantly decreased serum alanine aminotransferase, aspartate aminotransferase, and total bilirubin levels in D-GalN/LPS-challenged rats in a dose-dependent manner. The levels of enzymatic markers for oxidative stress (superoxide dismutase, glutathione peroxidase, catalase, and glutathione) were increased by UGBE treatment in a dose-dependent manner. Tumor necrosis factor alphalevel, inducible nitric oxide synthase activities, and nitric oxide productions were reduced by UGBE treatment. In addition, hemeoxygenase-1 levels in liver were also significantly increased in the UGBE-treated group. The protein expression of toll-like receptor 4 was decreased by UGBE administration. Hematoxylin and eosin staining results also supported the results of this study showing normal appearance of liver histopathology in the UGBE-treated group. Conclusion: UGBE showed a great hepatoprotective effect on D-GalN/LPS-challenged rats via the toll-like receptor 4 signaling pathway.

Healing and preventive effects of low-esterified pectin on liver injury induced by carbon tetrachloride in rats

  • Khotimchenko, Yuri S.;Kolenchenko, Elena A.;Khotimchenko, Maxim Y.;Kovalev, Valeri V.
    • Advances in Traditional Medicine
    • /
    • 제4권1호
    • /
    • pp.28-36
    • /
    • 2004
  • The purpose of this study was to investigate the pharmacological effects of low-esterified pectin on carbon tetrachloride $(CCL_4)-induced$ hepatotoxicity in rats. The study included two experiments. In the first experiment the animals were given daily $CCL_4$ through gavage for 7 days and then 10, 50, or 250 mg/kg b.w. of pectin for 21 days. At the end of experiment rats were killed within 24 hours. The increased bilirubin level, enhanced alanine aminotransferase and aspartate aminotransferase activity in plasma induced by $CCL_4$ were partly normalized by pectin administration in a dose-dependent manner. The pectin treatment also resulted in significant recovery of $CCL_4-induced$ decrease of the liver glycogen content. In addition, pectin significantly improved $CCL_4-induced$ alterations of pro-oxidant and antioxidant biochemical parameters in liver and plasma compared to those of rats administered $CCL_4$. In the second experiment the animals were given daily 10, 50 or 250 mg/ kg b.w. of pectin for 21 days before a 7-day administration of $CCL_4$. Rats were killed 24 hours after the end of experiment. Pretreatment with pectin before $CCL_4$ administration resulted in significantly inhibited increase of the blood enzymatic activities of alanine and aspartate aminotransferases and bilirubin level in a dose-dependent manner. Also, preliminary administration of pectin prevented elevation of malondialdehyde and conjugated diene levels in liver and plasma as well as a reduction of glutathione content in liver of rats given $CCL_4$. These results suggest that low-esterified pectin exert healing and preventive effects on $CCL_4-induced$ hepatotoxicity in rats.

구멍갈파래의 고압 균질 전처리 공정을 통한 바이오에탄올 생산용 당화수율 증진 (Enhancement of Saccharification Yield of Ulva pertusa Kjellman by High Pressure Homogenization Process for Bioethanol Production)

  • 최운용;이춘근;안주희;서용창;이상은;정경환;강도형;조정섭;최근표;이현용
    • KSBB Journal
    • /
    • 제26권5호
    • /
    • pp.400-406
    • /
    • 2011
  • This study was investigated to improve the saccharification yield of Ulva pertusa Kjellman by the high pressure homogenization process. It was found that the high pressure homogenization pretreatment effectively destructed the cell wall structures only by using water. The high pressure homogenization process was operated under various conditions such as 10000, 20000 or 30000 psi with different recycling numbers. The optimal condition was determined as 30000 psi and 2 pass of recycling numbers and the sugar conversion yields were 16.02 (%, w/w) of glucose and 14.70 (%,w/w) of xylose, respectively. In the case of enzymatic treating the hydrolyzates with 5 FPU/glucan of celullase and 100 units/mL of amyloglucosidase, 65.8% of carbohydrates was converted into glucose. Using the hydrolysates of Ulva pertusa Kjellman, 48.7% of ethanol was obtained in the culture S.cerevisiae. These results showed that the high pressure homogenization process could efficiently hydrolyze the marine resource by using only water for bioethanol production.

Nitric oxide modulates antioxidant defense and the methylglyoxal detoxification system and reduces salinity-induced damage of wheat seedlings

  • Hasanuzzaman, Mirza;Hossain, Mohammad Anwar;Fujita, Masayuki
    • Plant Biotechnology Reports
    • /
    • 제5권4호
    • /
    • pp.353-365
    • /
    • 2011
  • The present study investigates the possible regulatory role of exogenous nitric oxide (NO) in antioxidant defense and methylglyoxal (MG) detoxification systems of wheat seedlings exposed to salt stress (150 and 300 mM NaCl, 4 days). Seedlings were pre-treated for 24 h with 1 mM sodium nitroprusside, a NO donor, and then subjected to salt stress. The ascorbate (AsA) content decreased significantly with increased salt stress. The amount of reduced glutathione (GSH) and glutathione disulfide (GSSG) and the GSH/GSSG ratio increased with an increase in the level of salt stress. The glutathione S-transferase (GST) activity increased significantly with severe salt stress (300 mM). The ascorbate peroxidase (APX), monodehydroascorbate reductase (MDHAR), dehydroascorbate reductase (DHAR), catalase (CAT) and glutathione peroxidase (GPX) activities did not show significant changes in response to salt stress. The glutathione reductase (GR), glyoxalase I (Gly I), and glyoxalase II (Gly II) activities decreased upon the imposition of salt stress, especially at 300 mM NaCl, with a concomitant increase in the $H_2O_2$ and lipid peroxidation levels. Exogenous NO pretreatment of the seedlings had little influence on the nonenzymatic and enzymatic components compared to the seedlings of the untreated control. Further investigation revealed that NO pre-treatment had a synergistic effect; that is, the pre-treatment increased the AsA and GSH content and the GSH/GSSG ratio, as well as the activities of MDHAR, DHAR, GR, GST, GPX, Gly I, and Gly II in most of the seedlings subjected to salt stress. These results suggest that the exogenous application of NO rendered the plants more tolerant to salinity-induced oxidative damage by enhancing their antioxidant defense and MG detoxification systems.

Bioconversion of Lignocellulosic Materials with the Contribution of a Multifunctional GH78 Glycoside Hydrolase from Xylaria polymorpha to Release Aromatic Fragments and Carbohydrates

  • Liers, Christiane;Ullrich, Rene;Kellner, Harald;Chi, Do Huu;Quynh, Dang Thu;Luyen, Nguyen Dinh;Huong, Le Mai;Hofrichter, Martin;Nghi, Do Huu
    • Journal of Microbiology and Biotechnology
    • /
    • 제31권10호
    • /
    • pp.1438-1445
    • /
    • 2021
  • A bifunctional glycoside hydrolase GH78 from the ascomycete Xylaria polymorpha (XpoGH78) possesses catalytic versatility towards both glycosides and esters, which may be advantageous for the efficient degradation of the plant cell-wall complex that contains both diverse sugar residues and esterified structures. The contribution of XpoGH78 to the conversion of lignocellulosic materials without any chemical pretreatment to release the water-soluble aromatic fragments, carbohydrates, and methanol was studied. The disintegrating effect of enzymatic lignocellulose treatment can be significantly improved by using different kinds of hydrolases and phenoloxidases. The considerable changes in low (3 kDa), medium (30 kDa), and high (> 200 kDa) aromatic fragments were observed after the treatment with XpoGH78 alone or with this potent cocktail. Synergistic conversion of rape straw also resulted in a release of 17.3 mg of total carbohydrates (e.g., arabinose, galactose, glucose, mannose, xylose) per gram of substrate after incubating for 72 h. Moreover, the treatment of rape straw with XpoGH78 led to a marginal methanol release of approximately 17 ㎍/g and improved to 270 ㎍/g by cooperation with the above accessory enzymes. In the case of beech wood conversion, the combined catalysis by XpoGH78 and laccase caused an effect comparable with that of fungal strain X. polymorpha in woody cultures concerning the liberation of aromatic lignocellulose fragments.