• Title/Summary/Keyword: Enzyme injection

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Effects of Curcumae Longae Rhizoma Pharmacopuncture on Monosodium Iodoacetate-induced Osteoarthritis Rats (강황(薑黃) 약침이 Monosodium Iodoacetate 유도 골관절염 흰쥐에 미치는 영향)

  • Lee, Jong-Hoon;Woo, Chang-Hoon
    • Journal of Korean Medicine Rehabilitation
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    • v.29 no.2
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    • pp.115-133
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    • 2019
  • Objectives The purpose of this study was to evaluate the effects of Curcumae Longae Rhizoma pharmacopuncture on the monosodium iodoacetate (MIA)-induced osteoarthritis rats. Methods Osteoarthritis was induced by injection of MIA ($50{\mu}L$ with 80 mg/mL) into knee joint cavity of rats. Rats were divided into 6 groups. Normal group was injected by normal saline into knee joint cavity only. Control group was induced for osteoarthritis by MIA and orally administered with distilled water. Normal Saline group was induced for osteoarthritis by MIA and injected with normal saline $100{\mu}L$. Positive comparison group was injected with MIA and orally administered with indomethacin 5 mg/kg. Curcumae Longae Rhizoma pharmacopuncture low concentration (CL) group was induced for osteoarthritis by MIA and injected with Curcumae Longae Rhizoma pharmacopuncture low concentration $100{\mu}L$. Curcumae Longae Rhizoma pharmacopuncture high concentration (CH) group was induced for osteoarthritis by MIA and injected with Curcumae Longae Rhizoma pharmacopuncture high concentration $100{\mu}L$. Curcumae Longae Rhizoma pharmacopuncture was injected at ST35 and EX-LE4 each group (CL, CH). After that, hind paw weight distribution was measured and oxidative stress biomarker in serum, liver function biomarker in serum, western blot analysis were measured. Histological analysis of knee joint tissue was performed by hematoxylin and eosin staining, Safranin-O staining and Masson's trichrome staining. Results Hind paw weight distribution was significantly improved in both group. alanine aminotransferanse and aspartate aminotransferase were decreased significantly in CH group compare with Indomethacin threated group. Antioxidant enzyme glutathione peroxidase, Catalase and heme oxygenase-1 were increased in CH group compare with control group. Inflammatory cytokine cyclooxygenase-2, inducible nitric oxide synthase and interleukin-1 beta were decreased significantly in CH group. Histological analysis result shows that protective effects of joint and cartilage were observed in both CH and CL groups in a concentration-dependent. Conclusions The result suggest that Curcumae Longae Rhizoma pharmacopuncture has anti-oxidation effect, anti-inflammatory effect and also can prevent progression of osteoarthritis and protect joint cartilage.

Ahnak-knockout mice show susceptibility to Bartonella henselae infection because of CD4+ T cell inactivation and decreased cytokine secretion

  • Choi, Eun Wha;Lee, Hee Woo;Lee, Jun Sik;Kim, Il Yong;Shin, Jae Hoon;Seong, Je Kyung
    • BMB Reports
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    • v.52 no.4
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    • pp.289-294
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    • 2019
  • The present study evaluated the role of AHNAK in Bartonella henselae infection. Mice were intraperitoneally inoculated with $2{\times}10^8$ colony-forming units of B. henselae Houston-1 on day 0 and subsequently on day 10. Blood and tissue samples of the mice were collected 8 days after the final B. henselae injection. B. henselae infection in the liver of Ahnak-knockout and wild-type mice was confirmed by performing polymerase chain reaction, with Bartonella adhesion A as a marker. The proportion of B. henselae-infected cells increased in the liver of the Ahnak-knockout mice. Granulomatous lesions, inflammatory cytokine levels, and liver enzyme levels were also higher in the liver of the Ahnak-knockout mice than in the liver of the wild-type mice, indicating that Ahnak deletion accelerated B. henselae infection. The proportion of CD4+interferon-${\gamma}$ ($IFN-{\gamma}^+$) and $CD4^+$ interleukin $(IL)-4^+$ cells was significantly lower in the B. henselae-infected Ahnak-knockout mice than in the B. henselae-infected wild-type mice. In vitro stimulation with B. henselae significantly increased $IFN-{\gamma}$ and IL-4 secretion in the splenocytes obtained from the B. henselae-infected wild-type mice, but did not increase $IFN-{\gamma}$ and IL-4 secretion in the splenocytes obtained from the B. henselae-infected Ahnak-KO mice. In contrast, $IL-1{\alpha}$, $IL-1{\beta}$, IL-6, IL-10, RANTES, and tumor necrosis $factor-{\alpha}$ secretion was significantly elevated in the splenocytes obtained from both B. henselae-infected wild-type and Ahnak-knockout mice. These results indicate that Ahnak deletion promotes B. henselae infection. Impaired $IFN-{\gamma}$ and IL-4 secretion in the Ahnak-knockout mice suggests the impairment of Th1 and Th2 immunity in these mice.

Development of Voltammetric Nanobio-incorporated Analytical Method for Protein Biomarker Specific to Early Diagnosis of Lung Cancer (폐암 조기 진단을 위한 단백질 바이오마커 측정용 전압-전류법 기반의 나노바이오 분석법 개발)

  • Li, Jingjing;Si, Yunpei;Nde, Dieudonne Tanue;Lee, Hye Jin
    • Applied Chemistry for Engineering
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    • v.32 no.4
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    • pp.461-466
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    • 2021
  • In this article, a portable and cost-effective voltammetric biosensor with nanoparticles was developed for the measurements of heterogeneous nuclear ribonucleoprotein A1 protein (hnRNP A1) biomarker which can potentially be used for lung cancer diagnosis. Gold nanoparticles were first electrodeposited onto screen printed carbon electrode (SPCE) followed by immobilizing a single stranded DNA aptamer specific to hnRNP A1 onto the electrode surface. Ethanolamine was also used when immobilizing DNA aptamer on the surface to prevent signals from non-specific adsorption events. Sequential injection of hnRNP A1 biomarker and anti-hnRNP A1 conjugated with alkaline phosphatase (ALP) onto the aptamer chip surface allows to form the sandwich complex of DNA aptamer/hnRNP A1/ALP-anti-hnRNP A1 on the electrode surface which further reacted with 4-aminophenyl phosphate (APP). The electrocatalytic reaction of the enzyme, ALP, and the substrate, APP, resulting in the oxidative current response changes at -0.05 and -0.17 V (vs. Ag/AgCl) against the hnRNP A1 concentration was measured using cyclic and differential pulse voltammetry, respectively. The Au nanoparticles-integrated voltammetric biosensor was applied to analyze human normal serum solutions possibly suggesting potential applicability for lung cancer diagnosis.

Effects of Prunellae Spica Extract on LT4-induced Hyperthyroidism in Rats through the Regulation of Heat and Cold Imbalance (하고초추출물의 갑상선기능항진증 랫트모델에서의 한열조절작용에 의한 개선효능 연구)

  • Kang, An Na;Kang, Seok Yong;Meng, Xianglong;Ma, Junnan;Park, Jong Hun;Park, Yong-Ki
    • The Korea Journal of Herbology
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    • v.33 no.4
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    • pp.77-85
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    • 2018
  • Objective : This study was intended to examine the effects of water extract of Prunellae Spica (PS), which is a herb with 'cold' nature based on hot and cold theory of traditional Korean medicine. Methods : Hyperthyroidism was induced in SD rats by LT4 (0.5 mg/kg, i.p.) daily for four weeks. After 2 weeks of LT4 injection, rats were divided randomly into four groups; normal, LT4-induced hyperthyroid control, PS extract (500 mg/kg, p.o.)-treated group, and propylthiouracil (PTU, 10 mg/kg, s.c.)-treated positive group. After 2 weeks of drug treatment, all rats were sacrificed and harvested blood samples and thyroid tissues. The changes of body weight, food and water intake, and body temperature were measured weekly. Serological markers were analyzed in sera using an enzyme-based assay, and thyroid tissues were stained with Hematoxylin & Eosin (H&E). Brain and dorsal root ganglion (DRG) tissues were isolated and analyzed the expression of transient receptor potential (TRP) channels by Western blot. Results : PS extract administration attenuated the loss of body weight and the increase of body temperature in LT4-induced hyperthyroidism rats. PS extract increased the level of thyroid stimulating hormone (TSH) and decreased tiiodothyronine (T3) and tetraiodothyronine (T4). In action mechanism, PS extract regulated the expression of transient receptor potential channel subfamily V member 1 (TRPV1) and transient Receptor Potential channel subfamily M member 8 (TRPM8), the thermoregulators. Conclusion : To conclude, PS extract can improve the symptoms of hyperthyroidism through regulation of the thyroid hormones imbalance and thermoregulation via TRP channels.

The effects of Socheongryong-Tang on LPS-induced lung inflammation rats model (소청룡탕이 LPS로 유도된 폐손상 동물모델에 미치는 영향)

  • Jin, Bo-Ram;Choi, In Young;Hwang, Do Young;Ham, Seong-Ho;An, Hyo-Jin
    • The Korea Journal of Herbology
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    • v.34 no.5
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    • pp.21-28
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    • 2019
  • Objectives : In present study, we investigated a therapeutic effect and optimum dose of Socheongryong-Tang (SCT) on LPS-induced lung inflammation rats model. Methods : Male Sprague-Dawley rats ($260{\pm}10g$) were divided into 12 groups : Group 1 included the normal rats, and Group 2-12 were administrated LPS by intranasal injection to induce experimental lung inflammation. After 1 day of LPS administration, Group 3-9 were treated with SCT ${\times}1/4$, ${\times}1/2$, ${\times}1$, ${\times}3$, ${\times}6$, ${\times}12$ or ${\times}18$, respectively. Group 10-12 (positive control) were treated with dexamethasone 1 mg/kg or acetylcystein 1.5 mg/kg or diclofenac sodium 0.4 mg/kg, respectively. After sacrifice, bronchoalveolar lavage fluid (BALF) was isolated. The levels of IL-$1{\beta}$, TNF-${\alpha}$, mucin glycoprotein 5AC (MUG5AC) were measured in BALF using enzyme-linked immunosorbent assay (ELISA). Results : LPS injected rats exhibited outstanding lung inflammation manifestations, including increased amount of total cells and neutrophil, and upregulated inflammatory cytokines level in BALF. However, the administration of SCT ${\times}1/4$, ${\times}1/2$ and ${\times}1$ decreased total cells and neutrophil, and suppressed the production of inflammatory cytokines, including $IL-1{\beta}$ and TNF-${\alpha}$, and MUG5AC in BALF. Notably, inhibitory effect of SCT ${\times}1/2$ and ${\times}1$ on the level of TNF-${\alpha}$ was markedly better than that of positive controls, dexamethasone and acetylcystein. Conclusions : Taken together, these results suggest that SCT ${\times}1/2$ and ${\times}1$ has therapeutic effects on LPS-induced lung inflammation rats model.

Suppression of Microglial Activation by Acute Ethanol Administration through HT7 Stimulation (급성 알코올 투여 백서의 신문혈 자극이 소교세포 활성에 미치는 영향)

  • Su Yeon Seo;Se Kyun Bang;Suk Yun Kang;Seong Jin Cho;Kwang-Ho Choi;Yeonhee Ryu
    • Korean Journal of Acupuncture
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    • v.41 no.2
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    • pp.33-42
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    • 2024
  • Objectives : The sigma-1 receptor is implicated in stress, depression, psychostimulant sensitization, and addiction vulnerability. Prior studies have indicated that ethanol exposure modulates sigma-1 receptor activity within the Ventral Tegmental Area (VTA). Here, we explore the sub-mechanisms underlying sigma-1 receptor activity induced by HT7 (Shinmun) stimulation in behavioral alterations following acute ethanol (ETOH) administration. Methods : Male Wistar rats were investigated for pro- and anti-inflammatory markers after injection of ETOH (1 g/kg) using cytokine enzyme-linked immunosorbent assay (ELISA)s. After confirming that HT7 stimulation changed the total distance traveled in the open field test (OFT), protein changes in the Ventral tegmental area (VTA) were measured by Western blotting. The expression level of inducible nitric oxide synthase (iNOS) after administration of a sigma-1 receptor antagonist (dihydrobromide 1047; BD1047, 10 mg/kg i.p.) and Shenmen (HT7) stimulation was compared. Results : As a result, acute ETOH administration increased proinflammatory marker levels (TNF-𝛼 and IL-6). HT7 stimulation restored the total distance response after acute ethanol administration. In addition, in the VTA, the levels of a microglial marker (iNOS), sigma-1 receptor and protein kinase C, which are predicted to be involved in up- and downregulation, were restored by HT7 stimulation. In particular, HT7 stimulation modulates iNOS expression through effects similar to BD treatment. This study suggests that the stimulatory effect of HT7 may be driven by microglial activation. Conclusions : Microglial activity is regulated by sigma-1 receptor, and sigma-1 receptor activity is regulated by HT7 stimulation. Significantly, we demonstrate that HT7 stimulation ameliorates behavioral alterations induced by acute ETOH administration through microglial activation within the VTA.

Effect of Dietary Vitamin E Levels on Lipid Peroxidation and Enzyme Activities of Antioxidative System in Brain of Cadmium Administered Rats (식이 Vitamin E 수준이 Cadmium 중독된 흰쥐 뇌조직의 지질과산화와 항산화효소활성에 미치는 영향)

  • 정선영;이순재;양정아
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.25 no.4
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    • pp.575-580
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    • 1996
  • The purpose of this study was to investigate the effect of dietary vitamin E levels on the enzymes such as superoxide dismutase(SOD), glutathione peroxidase(GSH-Px) and glutathione S-transferase (G57) involved in antioxidative defense system and lipid peroxidation in brain of cadmium administered rats. Sprague-Dawely male rats weighing $60\pm5g$ were divided into control and experimental groups. The experimental groups were divided into Cd-0E(vitamin E free diet), Cd-40E(40mg vitamin E/kg diet) and Cd-400E(400mg vitamin E/kg diet) according to the level of vitamin E supplementation. After each group was fed diet ad libitum for 2 or 4 weeks, 2.5mg cadmium per kg body weight was injected intraperitoneally once a day for 4 days. The rats were sacrificed for examination on the next day after the last injection of cadmium. The results are as follows: SOD activities of rat brain were lower in Cd-0E, but had a similar tendency to Cd-40E, Cd-400E groups compared with control group. GSH-Px acivities of rat brain were decreased in Cd-400E, Cd-40E and Cd-0E groups. GST activities of rat brain were decreased in Cd-0E, Cd-40E groups and not significantly different in Cd-400E compared with control group. Thiobarbituric acid reactive substances(TBARS) of rat brain was increased in Cd-0E, Cd-40E, Cd-400E in that order, TBARS was lower in Cd-40E, and Cd-400E by 28.8% and 44%, respectively, than Cd-0E group. The present result suggests that high level of dietary vitamin E protects against lipid peroxidative damage in rat induced by cadmium.

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Superoxide Dismutase Gene Expression in the Endotoxin-Treated Rat Lung (내독소에 의한 백서 폐장의 Superoxide Dismutase 유전자 발현에 관한 연구)

  • Yoo, Chul-Gyu;Suh, Gee-Young;Kim, Young-Whan;Han, Sung-Koo;Shim, Young-Soo;Kim, Keun-Youl;Han, Yong-Chol
    • Tuberculosis and Respiratory Diseases
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    • v.41 no.3
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    • pp.215-221
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    • 1994
  • Background: It is well known that oxygen free radicals(OFR) play a vital role in the various type of acute lung injury. Among various antioxidant defense mechanisms, the superoxide dismutases(SOD) are thought to be the first line of antioxidant defense by catalyzing the dismutation of two superoxide radicals to yield hydrogen peroxide and oxygen. Eukaryotic cells contain two types of intracellular SOD : cytosolic, dimeric copper/zinc- containing enzyme(CuZnSOD) and mitochondrial, tetrameric manganese-containing enzyme(MnSOD). The purpose of this study is to evaluate the time-dependent gene expression of MnSOD and CuZnSOD in the endotoxin-treated rats, and to compare with the manifestations of LPS-induced acute lung injury in rats. Methods: Total RNA from rat lung was isolated using single step phenol extraction 0, 1, 2, 4, 6, 12, 18, 24 hours after E. coli endotoxin injection(n=3, respectively). RNA was separated by formaldehyde-containing 1.2% agarose gels elctrophoresis, transblotted, baked, prehybridized, and hybridized with $^{32}P$-labeled cDNA probes for rat MnSOD and CuZnSOD, which were kindly donated by Dr. Ho(Duke University, Durham, NC, USA). The probes were labeled by nick translation. Blots were washed and autoradiography were quantitated using laser densitometry. Equivalent amounts of total RNA/gel were assessed by monitoring 28S and 18S rRNA. Results: Endotoxin caused a rise in steady-state MnSOD mRNA levels by 4h with peak mRNA accumulation by 6h. Continued MnSOD mRNA expression was observed at 12h. CuZnSOD mRNA expression was observed from 1h to 24h with peak levels by 18h. Conclusion: These results suggest that SOD palys an important defensive role in the endotoxin-induced acute lung injury in rats.

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Effect of buchu (Allium tuberosum) on lipid peroxidation and antioxidative defense system in streptozotocin-induced diabetic rats (부추가 Streptozotocin 유발 당뇨쥐의 지질과산화와 항산화방어체계에 미치는 영향)

  • 송영선;정현실;노경희;조혜연;박지영;최춘연;권태완
    • Journal of Life Science
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    • v.13 no.3
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    • pp.333-342
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    • 2003
  • The pathogenic effort of high glucose, possibly in concert with fatty acids, is mediated to vascular complications of diabetes via increased production of reactive oxygen species(ROS), reactive nitrogen species(RNS), and subsequent oxidative stress. This study was carried out to investigate the suppressive effect of buchu(Allium tuberosum) on oxidative stress in streptozotocin(STZ)-induced diabetes in Sprague Dawley male rats. The effect of buchu supplementation (10%) on lipid peroxidation, and antioxidative defense system in blood and liver was compared among normal rats fed basal diet(normal) and diabetic rats fed basal diet(DM-control) or 10% buchu-supplemented diet(DM-buchu). Diabetes was experimentally induced by the femoral muscle injection of 50 mg STZ per kg of body weight. Animals were sacrificed after 4 wks of experimental diets feeding. The induction of diabetes by STZ elevated the level of lipid peroxidation represented by thiobarbituric acid-reactive substances(TBARS) and conjugated dienes in plasma, LDL, liver, and erythrocytes. 10% buchu-supplemented diet significantly reduced the levels of conjugated dienes in erythrocytes(p<0.05) and lowered TBARS in liver and LDL to the levels of control. Induction of diabetes by STZ elevated Mn-superoxide dismutase(Mn-SOD) activity and lowered activities of glutathionine reductase(GSH-red) and glutathionine peroxidase(GSH-px). Catalase activity was not affected by the induction of diabetes by STZ. However, buchu supplementation to diabetic rats significantly elevated catalase activity(p<0.05) and slightly elevated GSH-px and GSH-red activities in liver. GSH levels of blood and liver were lowered or not changed by induction of diabetes by STZ, respectively, while buchu supplementation to diabetic rats significantly elevated hepatic GSH level (p<0.05). In conclusion, it can be concluded that buchu might be a food source to attenuate oxidative stress in diabetic patients by inhibiting lipid peroxidation, by increasing hepatic GSH level, and by inducing anti-oxidative enzyme systems.

Cholinesterase Activities in Blood and Nervous Tissues of Rats following Intraperitoneal Repetitive Injection of Parathion (Parathion의 복강내 반복투여로 인한 Rat의 혈액 및 신경조직내 Cholinesterase 활성변화)

  • Do, Jae Cheul;Mo, Ki Chul;Kim, Young Hong;Huh, Rhin Sou
    • Current Research on Agriculture and Life Sciences
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    • v.6
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    • pp.171-180
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    • 1988
  • Parathion is widely used in agriculture, but it is highly toxic and now clear that parathion behaves like a cholinergic drug by inhibiting the enzyme cholinesterase. In order to know the effect of toxicity and cholinesterase activity in rats injected repeatedly with parathion, cholinesterase activity in plasma, whole brain and spinal cord, and the subacute toxicity after repetitive intraperitoneal injection of parathion 20 times every 3 days were investigated. The results obtained were summerized as follows ; $LD_{50}$ value of parathion given intraperitoneally to rats was 10.5mg/kg(95% confidence limits, 6.6-16.8mg/kg). In subacute toxicity test of parathion injected intraperitoneally, mortality of parathion-pretreated rats(B : 57%, C : 83%) were increased in comparison with the control(50%). Cholinesterase activities in plasma of parathion-pretreated rats(B : 0.47 U/ml, C : 0.36 U/ml, AA : 0.31 U/ml, B : 0.26 U/ml, CC : 0.17 U/ml) were significantly decreased in comparison with the control(0.58 U/ml). Cholinesterase activities in spinal cord of parathion-pretreated rats(B : 1.87 U/g, C : 1.29 U/g, AA : 1.27 U/g, BB : 0.71 U/g, CC : 0.25 U/g) were decreased in comparison with the control(2.48 U/g). Cholinesterase activities in whole brain of parathion-pretreated rats(B : 2.52 U/g, C : 1.32 U/g, AA : 2.48 U/g, BB : 1.08 U/g, CC : 0.51 U/g) were significantly inhibited in comparison with the control(4.67 U/g). However, there were no differences in the urea nitrogen and creatinine concentrations between parathion-pretreated rats and control.

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