• The Journal of Internal Korean Medicine
• /
• v.29 no.1
• /
• pp.200-218
• /
• 2008

Objectives : The purpose of this study was to investigate the effects of Injinchunggan-tang (Yinchenchinggan-tang) on DMN-induced liver damage by applying proteomics. Materials and Methods : Sprague-Dawley rats were used in this experiment and were divided into the normal group (normal saline), the control group (DMN) and the sample group (DMN+IJCGT). DMN was injected i.p. once a day three times a week for 3 weeks in the control group. Normal saline instead of DMN was administered to the normal group. In the sample group, Injinchunggan-tang (Yinchenchinggan-tang) extract was orally administered once a day for 10 days after DMN was induced. The livers of each group were processed and analyzed by histology, Western blot, $Oxyblot^{TM}$, CBB and 2-dimensional electrophoresis. Results : In the histological findings of the liver, IJCGT reduced collagen deposition and liver damage in DMN-induced hepatic fibrosis. IJCGT increased MMP-13 protein production assessed by western blot. Protein oxidation induced by DMN treatment was decreased by IJCGT. In the 2-dimensional electrophoresis finding, the level of the increased proteins induced by DMN treatment such as GRP 75, 58kDa glucose regulated protein and heat shock 70kDa protein 5 were decreased by IJCGT. IJCGT was considered to have the protective effects on hepatotoxicity induced by DMN. In the 2-dimensional electrophoresis finding, the level of increased oxidized proteins such as heat shock 70 protein, mitochondrial malonyltransferase, calreticulin precursor, actin, NADP-isocitrate dehydrogenase, ankyrin repeat and SOCS box protein 11 were decreased by IJCGT. IJCGT was considered to have protective effect on the protein production induced by DMN treatment. Conclusion : Injinchunggan-tang (Yinchenchinggan-tang) exerts an inhibitory effect against the fibrosis and protein oxidation induced by DMN treatment in the rat liver. IJCGT was considered to have protective effects on the hepatotoxicity and protein production induced by DMN treatment.

• Herbal Formula Science
• /
• v.21 no.2
• /
• pp.144-157
• /
• 2013

Objectives : The aim of this study was to evaluate the efficacy of Hwangnyeonhaedok-tang (HH) and Geongangbuja-tang (GB) on the plasma hormone level in mice exposed to cold stress. HH and GB are the representative prescriptions of cold and hot property, respectively. Methods : We established cold condition by confining ICR mice to a $4^{\circ}C$ cage for 24 hours, ICR mice were given a HH (100, 300, 1000 mg/kg) or GB (100, 300, 1000 mg/kg) extract orally twice a day for three consecutive days. From the second day, they were given cold stress ($4^{\circ}C$) for twenty four hours. To measure the plasma corticosterone, insulin, thyroxine, epinephrine and norepinephrine levels of mice, their blood samples were collected from cardiac puncture, immediately centrifuged at $4^{\circ}C$. The protein level of HSP70 and JNK was examined using western blot analysis in cortex and hypothalamus. Results : Oral administration of GB more significantly reduced plasma corticosterone level raised by cold stress than HH. Gardeniae Fructus (CJ), the constituent of HH, significantly increased the thyroxine level. Western blot analysis showed that cold stress-induced Heat shock protein 70 (HSP70) expression was increased by HH and GB, HH decreased JNK expression and GB increased JNK expression dose-depently in hypothalamus. Scutellariae Radix (HG), Zingiberis Rhizoma (GG) and Aconiti Tuber (BJ) decreased HSP70 in hypothalamus and GG, BJ decreased HSP70 in cortex as well. Conclusions : These results suggest Geongangbuja-tang (GB) is more effective for ameliorating the stress response caused by cold stress.

• Korean Journal of Microbiology
• /
• v.38 no.2
• /
• pp.67-73
• /
• 2002

The cellular responses of soil-borne bacterium, Pseudomonas sp. HK-6 to explosive 2,4,6-trinitrotoluene (TNT) were examined. Two stress shock proteins (SSPs), approximately 70-kDa DnaK and a 60-kDa GroEL were found in HK-6 cells in response to TNT. Analyses of SDS-PAGE and Western blot using anti-DnaK and GroEL revealed that SSPs were induced in HK-6 cells exposed to 0.5 M of TNT far 6-12 hrs. The maximum induction of proteins was achieved at 8-hr incubation point after HK-6 cells'exposure to TNT. Similar SSPs were found to be induced in HK-6 cells by heat shock (shift of temperature, from $30^{\circ}C$ to $42^{\circ}C$) or cold shock (shift of temperature,$30^{\circ}C$ to $4^{\circ}C$).2D-PAGE of soluble protein tractions from the culture of Pseudomonas sp. HX-6 exposed to TNT demonstrated that approximately 450 spots were observed on the silver stained gels ranging from pH 3 to pH 10. Among them, 12 spots significantly induced and expressed in response to TNT were selected and analyzed. Approximately 60-kDa protein, which was assumed highly expressed on the gel, was used for amino acid sequencing. N-terminal microsequencing with in-gel digestion showed that N-terminal sequence of the TNT-induced protein, <$^1XXAKDVKFGDSARKKML^17$, shared extensive similarity with $^1XXAKDVKFGDSARKKML^17$, N-terminal sequence of (P48216) GroEL of Pseudomonas putida.

• Journal of Periodontal and Implant Science
• /
• v.28 no.1
• /
• pp.103-120
• /
• 1998

Prokaryotic and eukaryotic cells respond to heat stress and other environmental abuses by synthesizing a small set of stress proteins and by inhibiting post-transcription synthesis of normal proteins. The purpose of the present study was to document the stress response produced by inflamed gingival tissue in vivo, and cytokine inducted human periodontal ligament cells. Human PDL cells were exposed to TNF-$\alpha$(1ng/ml), INF-$\gamma$(200 U/ml), LPS(100ug/ml), combination of cytokine, and SDS-PAGE gels running and Western blotting analysis was done. In vivo studies, the healthy gingival tissusse of a control group and inflamed gingival tissue of adult periodontitis were studied by immunohistochemistry and histology. The results were as follows 1. HSP 47 was distributed on basal layer in healthy gingiva, but stronger stained in basal, suprabasal, and spinous layer of inflamed gingiva. 2. HSP 47 was rare on endothelial cells and mononuclear cells in healthy gingiva, but stronger expressed in inflamed gingiva. 3. HSP 70 expression was rare on epihelium and inflammatory cells hi both healthy & inflamed gingiva. 4. HSP 70 was actively expressed on endothelial cells and inflammatory cells of capillary lumen in moderately & mild inflamend gingiva. 5. PDL cells showed low level of HSP 47 protein expression which was significantly induced by cytokine stimulation (LSP only and combination). 6. Maximum HSP 70 protein induction was seen with stimulation by a combination of the cytokine, Combination of TNF-$\alpha$, INF-$\gamma$, LPS have been shown to synergistically effects of HSP 70 expression. On the above findings, HSP Is influenced by cytokine and chronic inflammation in vivo, and may be involved in protection of tissue during periodontal inflammatiom.

• Journal of Acupuncture Research
• /
• v.24 no.6
• /
• pp.29-36
• /
• 2007

Electroacupuncture(EA) is known to affect various autonomic functions such as blood pressure regulation, immune modulation and the improvement of disorders concerning autonomic functions. The aim of the present study was to establish whether EA has an immune-enhancing effect. ICR mice weighing 20 to 25g were divided into four groups: Group I(n=6), blank; Group II(n=6), control; Group III(Zusanli, n=6), low frequency-EA(10Hz of electrical stimulation), and Group IV(Zusanli, n=6), high frequency-EA (100Hz of electrical stimulation). For this study, we investigated expressions of spleen heat shock protein (HSP)70, HSP90 and secretions of cytokines. A Forced Swimming Test(FST) was performed as a model of activity test in mice. After three days of the FST, 10Hz EA($114.8{\pm}7.27s$) and 100Hz EA($147.5{\pm}1.29s$) immobility time significantly decreased compared with the control group($157.2{\pm}1.48s$). After seven days, 10Hz EA($124{\pm}1s$) and 100Hz EA($141{\pm}4.24s$) also significantly decreased immobility time compared with the control group($168{\pm}7.93s$). 10Hz EA and 100Hz EA increased the expression of HSP70 but did not change that of HSP90. 100Hz EA increased secretions of IL-6 and IL-12 compared with the control group however, 10Hz EA failed to change those of IL-6 and IL-12. The present results suggest that EA may be useful for down-regulated immune diseases.

• Asian-Australasian Journal of Animal Sciences
• /
• v.30 no.2
• /
• pp.246-253
• /
• 2017

Objective: Present study explores the effect of hot summer period on the glycolytic rate of early post-mortem meat quality of Ghungroo and Large White Yorkshire (LWY) pig and comparative adaptability to high temperature between above breeds by shifting the expression of stress related genes like mono-carboxylate transporters (MCTs) and heat shock proteins (HSPs). Methods: Healthy pigs of two different breeds, viz., LYW and Ghungroo (20 from each) were maintained during hot summer period (May to June) with a mean temperature of about $38^{\circ}C$. The pigs were slaughtered and meat samples from the longissimus dorsi (LD) muscles were analyzed for pH, glycogen and lactate content and mRNA expression. Following 24 h of chilling, LD muscle was also taken from the carcasses to evaluate protein solubility and different meat quality measurements. Results: LWY exhibited significantly (p<0.01) higher plasma cortisol and lactate dehydrogenase concentration than Ghungroo indicating their higher sensitivity to high temperature. LD muscle from LWY pigs revealed lower initial and ultimate pH values and higher drip loss compared to Ghungroo, indicating a faster rate of pH fall. LD muscle of Ghungroo had significantly lower lactate content at 45 min postmortem indicating normal postmortem glycolysis and much slower glycolytic rate at early postmortem. LD muscle of LWY showed rapid postmortem glycolysis, higher drip loss and higher degrees of protein denaturation. Ghungroo exhibited slightly better water holding capacity, lower cooking loss and higher protein solubility. All HSPs (HSP27, HSP70, and HSP90) and MCTs (MCT1, MCT2, and MCT4) in the LD muscle of pigs inclined to increase more in Ghungroo than LWY when exposed to high temperature. Conclusion: Effect of high temperature on the variation of HSPs and MCTs may play a crucial role in thermal tolerance and adaptation to different climatic conditions, pH regulation, muscle acidification, drip loss, protein denaturation and also in postmortem meat quality development.

• Weed & Turfgrass Science
• /
• v.4 no.3
• /
• pp.256-261
• /
• 2015

Trampling stress in turfgrass fields caused by traffics often occurs in zoysiagrass (Zoysia japonica) fields including golf course. In order to know the influences of trampling stress on the growth and development of turfgrass, leaf and root growth, chlorophyll fluorescence, chlorophyll content and 2-DE protein analysis were conducted in the turfgrass plants subjected to various levels of trampling stress from 0 to $9,420J\;day^{-1}$ day. Shoot growth was more highly inhibited by trampling stress than root growth. Although root growth was affected by trampling with weak intensity, the highest root length was observed in the turfgrass treated with weak trampling ($1,570J\;day^{-1}$). Chlorophyll fluorescence (Fv $Fm^{-1}$) was drastically lowered by trampling stress with moderate intensity. Leaf number showed similar tendency with leaf greenness. The number was decreased as the trampling intensity was increased. Shoot dry weight was decreased showing a similar tendency with the result of shoot length. The specific protein expressions under weak trampling were related to the functions of stress amelioration. Heat shock 70 kDa protein 10 and oxygen-evolving enhancer protein 1 were the proteins increased positively under trampling stress.

• Radiation Oncology Journal
• /
• v.28 no.2
• /
• pp.91-98
• /
• 2010

Purpose: Troglitazone (TRO), a PPAR-$\gamma$ agonist, can reduce heat shock protein (HSP) 70 and increase the antioxidant enzymes, such as superoxide dismutase (SOD) and catalase, which might affect thermal sensitivity. Here, we investigated whether TRO modifies thermal sensitivity in uterine cervical cancer cells, which is most commonly treated by hyperthermia (HT). Materials and Methods: HeLa cells were treated with $5{\mu}M$ TRO for 24 hours before HT at $42^{\circ}C$ for 1 hour. Cell survival was analyzed by clonogenic assay. The expression of HSPs was analyzed by Western blot. SOD and catalase activity was measured and reactive oxygen species (ROS) was measured using 2',7'-dichlorofluorescin diacetate and dihydroethidium. Results: The decreased cell survival by HT was increased by preincubation with TRO before HT. Expression of HSP 70 was increased by HT however, it was not decreased by preincubation with TRO before HT. The decreased Bcl-2 expression by HT was increased by preincubation with TRO. SOD and catalase activity was increased by 1.2 and 1.3 times,respectively with TRO. Increased ROS by HT was decreased by preincubation with TRO. Conclusion: TRO decreases thermal sensitivity through increased SOD and catalase activity, as well as scavenging ROS in HeLa cells.

• Journal of the Korean Society of Physical Medicine
• /
• v.8 no.1
• /
• pp.111-116
• /
• 2013

연구목적: 본 연구는 흰쥐를 대상으로 정강뼈의 관절연골에 적용한 맥동전자장과 맥동초음파가 HSP70(Heat shock protein 70)의 발현을 통한 연골 형성에 미치는 영향을 알아보고자 실시하였다. 연구방법: 36마리의 200~250g의 Sprague-Dawley 흰쥐를 대조군, 맥동전자장 적용군, 맥동초음파 적용군으로 각 집단별로 12마리씩 무작위 배정하여 실험을 진행하였다. 맥동전자장은 27.12 MHz의 주파수, 5가우스의 강도, 450 W의 출력으로 10분간 적용하였고, 맥동초음파는 20%의 맥동비, 1MHz의 주파수, $1.5W/cm^2$의 강도로 10분간 적용하였다. 연구결과: 맥동전자장 적용군과 맥동초음파 적용군의 관절연골 조직에서 유의한 수준의 HSP70 발현량을 나타냈다. 또한 맥동전자장 적용군과 맥동초음파 적용군에서는 Akt, Erk1, CREB의 높은 활성도를 나타내었고, 맥동초음파 적용군에 비해서 맥동전자장 적용군의 더 높은 수준의 활성도를 보였다. 결론: 맥동전자장과 맥동초음파는 HSP70의 과발현을 유발하고, 이를 통해 연골형성을 증가시키는 것으로 나타나, 향후 관절연골의 손상에 대한 임상적 적용을 위한 추가적인 연구가 진행되어야 할 것으로 생각된다.

• BMB Reports
• /
• v.55 no.10
• /
• pp.488-493
• /
• 2022

The specific pair of heat shock protein 70 (Hsp70) and Hsp40 constitutes an essential molecular chaperone system involved in numerous cellular processes, including the proper folding/refolding and transport of proteins. Hsp40 family members are characterized by the presence of a conserved J-domain (JD) that functions as a co-chaperone of Hsp70. Tumorous imaginal disc 1 (Tid1) is a tumor suppressor protein belonging to the DNAJA3 subfamily of Hsp40 and functions as a co-chaperone of the mitochondrial Hsp70, mortalin. In this work, we performed nuclear magnetic resonance spectroscopy to determine the solution structure of JD and its interaction with the glycine/phenylalanine-rich region (GF-motif) of human Tid1. Notably, Tid1-JD, whose conformation was consistent with that of the DNAJB1 JD, appeared to stably interact with its subsequent GF-motif region. Collectively with our sequence analysis, the present results demonstrate that the functional and regulatory mode of Tid1 resembles that of the DNAJB1 subfamily members rather than DNAJA1 or DNAJA2 subfamily proteins. Therefore, it is suggested that an allosteric interaction between mortalin and Tid1 is involved in the mitochondrial Hsp70/Hsp40 chaperone system.