This study was performed for searching for non-hepatectomy medium-term bioassay model by using newborn female rats. Newborn female Sprague-Dawley rats (1 day old) were given an intraperitoneal injection of 150 mg/kg of diethylnitrosamine (DENA). After three weeks, all rats were weaned and divided into three groups. Group 1 were fed on diets containing 0.01% 2-acetylaminofluorene (2-AAF) as a promoter for three weeks. Group 2 were given 0.05% phenobarbital (PB) in drinking water as a promoter for 8 weeks. Group 3 was control group. The autopsy was carried out at 4 weeks and 8 weeks after weaning. Preneoplastic lesions were indentified with immunohistochemical staining for glutathione S-transferase placental form (GST-P). In liver weight to body weight ratios, group 2 showed significant difference from group 1 (p<0.001) at 4 weeks after weaning. Group 1 and group 2 showed significant difference from group 3 at 8 weeks after weaning (p<0.0I, p<0.001), respectively. In quantitative analysis for GST-P positive lesion area by using Image Analyzer, group 1 and group 2 represented significant difference in comparison with group 3 at early 4 weeks after weaning (p
Objectives : The usage of acupuncture has gained popularity for certain chronic pain conditions. However, the efficacy of acupuncture in various diseases has not been fully established and the underlying mechanism is not clearly understood. In the present study, the effect of electroacupuncture (EA) applied to yangno$(SI_6)$ on the neuropathic pain was examined. Methods : A common source of persistent pain in human is a neuropathic pain. Neuropathic pain was induced by tight ligation of L5 spinal nerve. When rats developed pain behaviors, EA was applied for 30 min. under enflurane anesthesia with repeated train stimuli at the intensity of 10X of muscle twitch threshold. The foot withdraw latency of the hind limb was measured for an indicator of pain level after each manipulation. Results : EA increased the mechanical threshold of the foot in the rat model of neuropathic pain significantly for the duration of 1 hr. suggesting a partial alleviation of pain. EA applied to SI6 point produced a significant improvement of mechanical sensitivity of the foot lasting for at least 1 h. However, $ST_{36}$ point did not produce any significant increase of mechanical sensitivity. The improvement of mechanical threshold was interpreted as an analgesic effect. The analgesic effort was specific to the acupuncture point since the analgesic effect on the neuropathic pain model could not be mimicked by EA applied to a point, $ST_{36}$. In addition, this analgesic effect of EA is mediated by a adrenergic mechanism of descending control of spinal cord from the brain. Conclusions : The data suggest that EA produces a potent analgesic effect on the neuropathic pain model in the rat; and 2) that EA-induced analgesia is mediated by a adrenergic mechanism of descending control in a point specific manner.
Objectives : DaeBangPungTang(DBPT) is one of the prescriptions used for the treatment of knee arthritis in oriental medicine. The present study aimed to examine the analgesic effect of DBPT on a rat model of carrageenan-induced arthritis, and the relations between DBPT-induced analgesia and endogenous nitric oxide(NO) and inducible NO synthase (iNOS), cyclooxygenase-2 (COX-2), and c-Fos protein expression in the spinal cord. Methods : Carrageenan-induced arthritis rat model was used to test the effect of DBPT as a chronic pain model. After the induction of arthritis, rats subsequently showed a reduced stepping force of the affected limb for at least tile next 4 days. The reduced stepping force of the limb was presumably due to a painful knee, since oral infection of indomethacin produced temporary improvement of weight bearing. DBPT dissolved in normal saline was minted several acupoints. After the treatment, behavioral tests measuring stepping force were periodically conducted during the next 12 hours. Results : DBPT produced significant improvement of stepping force of the hindlimb affected by the arthritis lasting at least 9 hours. The magnitude of this improvement was equivalent to that obtained after an oral injection of 3mg/kg of indomethacin and this improvement of stepping force was interpreted as an analgesic effect. DBPT produced the improvement of stepping force of the affected hindlimb in a dose-dependent manner. Both NO production and iNOS, COX-2 protein expression increased by arthritis were suppressed by DBPT. DBPT on combination with electroacupuncture (EA) produced more powerful and longer lasting improvement of stepping force of the hindlimb affected by the arthritis than either DBPT or EA did. Conclusion : The present study suggest that DBPT produces a potent analgesic effect on the chronic hee arthritis pain model in the rat and that DBPT-induced analgesia modulate endogenous NO through the suppression of iNOS/COX-2 protein expression.
Obiecnves : The purpose of this study was to examine the analgesic effect of moxi-tar on a rat model of carrageenan-induced arthritis and the relations between moxi-tar-induced analgesia and endogenous NO and iNOS, cyclooxygenase-2 (COX-2), and c-Fos protein expression in the spinal cord. Methods : Carrageenan-induced arthritis rat model was used to test the effect of moxi-tar as a chronic pain model. After the induction of arthritis, rats subsequently showed a reduced stepping force of the affected limb for at least the next 4 days. The reduced stepping force of the limb was presumably due to a painful knee, since oral injection of indomethacin produced temporary improvement of weight bearing. Moxi-tar dissolved in ethyl alcohol was injected several acupoints. After the treatment, behavioral tests measuring stepping force were periodically conducted during the next 12 hours. Results : Moxi-tar produced significant improvement of stepping force of the hindlimb affected by the arthritis lasting at least 9 hours. The magnitude of this improvement was equivalent to that obtained after an oral injection of 3 mg/kg of indomethacin and this improvement of stepping force was interpreted as an analgesic effect. Moxi-tar produced the improvement of stepping force of the affected hindlimb in a dose-dependent manner. Both NO production and iNOS, COX-2 protein expression increased by arthritis were suppressed by moxi-tar. moxi-tar on combination with electroacupuncture (EA) produced more powerful and longer lasting improvement of stepping force of the hindlimb affected by the arthritis than either moxi-tar or EA did. Conclusion : The present study suggest that moxi-tar produces a potent analgesic effect on the chronic knee arthritis pain model in the rat and that moxi-tar-induced analgesia modulate endogenous NO through the suppression of iNOS/COX-2 protein expression.
Objectives: This study was produced to examine the effects of moxibustion that had been played important role to traditional oriental medical treatment on disease. Recently, it was reported that moxi-tar which is generated in the process of moxibustion as burning combustibles decreased nitric oxide(NO) and inducible NO synthase (iNOS) generation in cellular experiments. Methods: Carrageenan-induced arthritis rat model was used to test the effect of moxi-tar as a chronic pain model. Diluted moxi-tar was single injected in several acupoints or combined with electroacupuncture (l ms, 2 Hz, and 2 mA) into contralateral ST36 acupoint for 30 min to assess the synergic effects. After the treatment, behavioral tests measuring stepping force were periodically conducted during the next 12 hours. Endogenous NO and iNOS, cyclooxygenase-2 (COX-2), and c-Fos protein expression in the spinal cord were examined on a rat model of carrageenan-induced arthritis. Results : After the induction of arthritis, rats subsequently showed a reduced stepping force of the affected limb for at least the next 4 days. The reduced stepping force of the limb was presumably due to a painful knee, since oral injection of indomethacin produced temporary improvement of weight bearing. Maxi-tar produced significant improvement of stepping force of the hindlimb affected by the arthritis lasting at least 9 hours. The magnitude of this improvement was equivalent to that obtained after an oral injection of 3 mg/kg of indomethacin and this improvement of stepping force was interpreted as an analgesic effect. Maxi-tar produced the improvement of stepping force of the affected hindlimb in a dose-dependent manner. Both NO production and iNOS, COX-2 protein expression increased by arthritis were suppressed by maxi-tar. Moxi-tar on combination with electroacupuncture (EA) produced more powerful and longer lasting improvement of stepping force of the hindlimb affected by the arthritis than either moxi-tar or EA did. Conclusion : The present study suggest that maxi-tar produces a potent analgesic effect on the chronic knee arthritis pain model in the rat and that moxi-tar-induced analgesia modulate endogenous NO through the suppression of iNOS/COX-2 protein expression.
Placenta-derived mesenchymal stem cells (PD-MSCs) are promising candidates for cell-based therapy in regenerative medicine. The migration and homing potential of PD-MSCs to injured sites is a critical property of MSC engraftment. MicroRNAs (miRNAs) have recently been shown to regulate the critical functions of MSCs, such as proliferation, survival, and migration. The objective of the present study was to identify the miRNA and target genes involved in PD-MSCs homing in a bile duct ligation (BDL) rat model. We selected candidate miRNAs targeting genes for PD-MSCs homing based on microarray analysis. PD-MSC engraftment in BDL-injured rat liver was identified by immunofluorescence assay and human-specific Alu gene expression by quantitative real-time polymerase chain reaction (qRT-PCR) one week after transplantation. Compared with migrated naïve PD-MSCs under hypoxic and normoxic conditions (Hyp/Nor), the transplanted group with PD-MSCs (Tx) showed distinct differences in miRNA expressions in BDL-injured rat liver. We also validated the miRNAs and their target genes for PD-MSCs homing. The expressions of integrin α4 (ITGA4) and integrin α5 (ITGA5) target genes for miR-199a-5p and miR-148a-3p were significantly upregulated in the Tx group (p<0.05). In addition, integrin β1 (ITGB1) and integrin β8 (ITGB8) were upregulated by suppressing miR-183-5p and miR-145-5p, respectively. These results demonstrated that PD-MSCs regulate miRNA expression related to the integrin family for their homing effects on the BDL-injured rat liver. The findings further suggest that miRNA-mediated regulation of the integrin family contributes to the therapeutic efficacy of PD-MSCs in the rat hepatic fibrosis model by BDL.
Fifty percent ethanol extract of Lythrum salicaria Linne root (LSR) was tested in vitro on antioxidant activity, and furthermore was investigated on antioxidative and fibrosis protecting activities in $CCL_4$-induced liver fibrosis rat model. Ratio of hepatic GSH/GSSG (reduced glutathione/oxidized glutathione) as bio-parameter of antioxidant level in $CCL_4$ plus LSR-treated rats for 6 weeks significantly increased from 2.8- to 5.7-fold than that of $CCL_4$-treated rats at p < 0.05. Thiobarbituric acid reactive substances (TBARS) contents in $CCL_4$ plus LSR-treated rats ranged from 1.57- to 2.19-fold of normal rats and were lower than those in $CCL_4$ plus silymarin-treated rats ($1.78{\sim}2.46$-fold of normal rats) (p < 0.05). Amounts of hydroxyproline of liver tissue showing the content of total collagen, a parameter of fibrosis, in $CCL_4$ plus LSR-administrated rat livers were $4.9{\sim}8.8{\mu}g$/mg ($-47{\sim}-71%$, compared with that in $CCL_4$-treated rat livers ($16.6{\mu}g$/mg tissue), which were significantly lower than those in $CCL_4$ plus silymarin-administrated rats being $8.4{\sim}11.7{\mu}g$/mg ($-30{\sim}-50%$). This collagen reducing effect of liver tissue in $CCL_4$ plus LSR-treated rats was supported by histological observation using microscopy assay. From the results, we conclude that the root of L. salicaria have efficient antioxidant potential and effective antifibrotic activities.
Objectives: Cinnamomi Ramulus (CR), the young twig of Cinnamomum loureirri nees, has been used for treating symptoms related to pain, rheumatic arthritis and inflammation in Korean herb medicine. This study was carried out to investigate the anti-inflammatory effect of CR in vivo and in vitro. Methods: Extracts of CR were prepared and the chemical components of the extracts were examined by gas chromatography-mass spectrometry (GC-MS). The extracts were administrated to the rat paw edema model induced by carrageenan to evaluate the anti-inflammatory effect of CR. The expressions of nitric oxide (NO), prostaglandin E2 (PGE2), inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX)-2 were also quantified in lipopolysaccharide(LPS)induced RAW 264.7 macrophages to survey the effect of CR in vitro. The main components were cinnamaldehyde and coumarin. Results: We examined the anti-inflammatory activity of the $80\%$ ethanol extract of Cinnamomi Ramulus in vivo by using carrageenan-induced rat paw edema model. Maximum inhibition of $54.91\%$ was noted at the dose of l1000mg/kg after 2 hours of drug administration in carrageenan-induced rat paw edema and this showed a potent anti-inflammatory effect. Conclusions: The results showed that Cinnamomi Ramulus suppressed dose-dependently LPS-induced NO production in RAW 264.7 macrophages and also decreased iNOS protein expression. Cinnamomi Ramulus also showed a significant inhibitory effect in LPS-induced PGE2 production and COX-2 expression.
Kim, Ji-Sook;Shim, Hyung-Jin;Kim, Hong-Jin;Choi, Kyung-Hee;Kim, Jung-Woong;Kwak, Byung-Kook
Biomedical Science Letters
/
v.13
no.3
/
pp.207-212
/
2007
The purpose of this preliminary study is to improve the efficiency of gene transfer of nonviral plasmid DNA by in vivo electroporation in ischemic hindlimb muscle, tibialis anterior. Hindlimb ischemic model was aseptically made by excision of left femoral artery. Each $50\;{\mu}g$ of pEGFP-C1 and pGL3-control in $100\;{\mu}l$ 0.9% NaCl was injected in tibialis anterior muscle. In vivo electroporation was applied on the same site with 10 mm-distance 2 needle array electrodes and ECM830. In 3 groups of normal rat with different electric field strength 0, 200 and 800 V/cm, the expression of pEGFP-C1 was comparatively evaluated. In 8 groups of normal rats, the expression of pGL3-control was evaluated in 0, 40, 50, 80, 100, 150, 200 and 300 V/cm of electric field strength. In 5 groups of ischemic models, the expression of pGL3-control was analyzed on 0, 4, 7, 10 and 14 days elapsed after making ischemic models. In 9 groups of ischemic rats, the expression of pGL3-control was analyzed in the electric field strength 0, 60, 70, 80, 100, 150, 200, 250 and 300 V/cm. GFP expressions in normal tibialis anterior were high in the extent and degree in order of electric field strength of 200, 800 and 0 V/cm. Luciferase value was highest in $50{\sim}100\;V/cm$ electric field strength. In the case of ischemic models, luciferase expression was significantly increasing in the order elapsed time after making the model. The degree of luciferase expression was higher in cases of application of in vivo electroporation than in that of non-application and was highest in $100{\sim}150\;V/cm$. In conclusion, in vivo electroporation is effective in transfer and expression of plasmid DNA in normal and ischemic tibialis anterior of rat.
Park Dong-Wan;Kim Wan-Sik;Bae Cheol-hwan;Jeong Sung-Hyun;Shin Gil-Cho;Lee Won-Chul
The Journal of Korean Medicine
/
v.25
no.3
/
pp.123-136
/
2004
Objectives : The purpose of this investigation is to evaluate the effects of Woohwangcheongsim-won (WC) on the in vitro neuronal development and alteration in gene expression in a hypoxia model using cultured rat cortical cells. Methods : E/sub 18/ rat cortical cells were grown in a neurobasal medium containing B27 supplement and various concentration of WC. Initial development of growth cone was investigated by phase-contrast microscopy, while dendritic spine formation and synaptogenesis were investigated by immunocytochemistry with SynGAPα(a postsynaptic marker) and synaptophysin (presynaptic marker) antibodies. Alteration in gene expression was analyses by microarray using rat 5K-TwinChips. Results : WC suppressed the development of growth cones and WC increased the number of dendritic spines at 20 and 50㎍/mL concentration but there was no statistical significance. Instead, it significantly decreased the number at 100㎍/mL. The expression of anti-apoptosis gene Bcl2-like 1 (Bcl211) increased (Global M=0.46), while Akt1 decreased. Proapoptosis genes Bad and PDCD2 increased. The expression of hemoglobin alpha 1 (probably neuroglobin) increased (Global M=0.93). The expression of antioxidants such as catalase, heme oxygenase (HO), and PRKAG2 gene increased. The expression PKC gene increased. The expression of retinoic acid receptor alpha (RARα) increased significantly (Global M=1.0). Conclusions : These data suggest that WC trends to suppress cellular activity slightly in normoxia and increases the expression of apoptosis-, antioxidation-, oxygen capture-related genes in hypoxia, but increases Bcl111 that anti-apoptosis gene, on the other hand increases Bad, PDCD2 that pro-apoptosis genes, too..
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