Ischemic stroke is a high mortality disease that causes irreversible damage. Chlorogenic acid is a polyphenolic substance with neuroprotective properties. Bcl-2 family proteins perform a critical role in apoptosis process. Bcl-2 and Bcl-xL are anti-apoptotic proteins that prevent cell death, and Bax and Bad are pro-apoptotic proteins that promote apoptosis. We investigated whether chlorogenic acid modulates Bcl-2 family proteins during focal cerebral ischemia. We made a rat model of ischemic stroke by performing middle cerebral artery occlusion (MCAO). Chlorogenic acid (30 mg/kg) or phosphate-buffered saline was treated via intraperitoneal injection 2 hr before MCAO. Neurological behavioral tests were performed 24 hr after MCAO damage and cortical tissues were collected. Reverse transcription-PCR, Western blot, and immunofluorescence staining were performed to observe changes in Bcl-2 family proteins expression. MCAO-damage induced neurobehavioral disorders and chlorogenic acid alleviate these deficits. Bcl-2 and Bcl-xL expressions were decreased and Bax and Bad expressions were increased in MCAO animals. However, chlorogenic acid treatment attenuated the decrease of Bcl-2 and Bcl-xL and the increase of Bad and Bax due to MCAO surgery. Moreover, chlorogenic acid treatment attenuated MCAO-induced upregulation of caspase-3. These findings suggest that chlorogenic acid exerts neuroprotective effects against MCAO damage by regulating Bcl-2 family proteins including Bcl-2, Bcl-xL, Bax, and Bad.

Traditional medicine and herbal remedies are gaining popularity worldwide, comprising a significant portion of healthcare research, advancements, and market demand. Growing scientific evidence supports their substantial efficacy as pharmaceutical ingredients and dietary supplements in preventive healthcare. When developing pharmaceuticals, it is crucial to ensure that ingredients are free from side effects and toxicity in order to prioritize safety. Geckos, known as shou gong, are a diverse group of lizards that are widely utilized for treating various diseases in Korean Medicine. This study was conducted to assess the potential acute toxicity of a water extract Gekko gecko by a single oral dose in Sprague-Dawley rats. Twenty rats of each sex were randomly assigned to four groups (5 rats each). Test articles were administrated once by oral gavage to rats at dose levels of 0, 500, 1,000, or 2,000 mg/kg body weight. Mortality, changes of body weight, and clinical signs of gross observation were monitored for 14 days after dosing. At the end of a 14-day observation period, all animals were sacrificed and complete macroscopic and hematological examinations were performed. There was no dead animal or test article-related effect on clinical signs, body weight, or gross finding. Other specific changes were not found between control and treated groups in hematology. Results showed no adverse effect at a dose of 500, 1,000, or 2,000 mg/kg in rats. The minimal lethal dose was considered to be over 2,000 mg/kg body weight in rats.

GN01 is a new antiviral medicine acting against Korean Sacbrood virus (KSBV) of honeybees. It contains 5 mg/mL of active ingredient, double stranded RNAs(dsRNA), that homologous to KSBV ribonucleic acid coding coat protein (VP1) of virus and inducing RNA interference (RNAi). RNA medicine is generally recognized as safe for rapid breakage by intrinsic ribonuclease and limited absorption from gastrointestinal tract. However, there were no data of repeat-dose toxicity in laboratory animals for dsRNA targeting SBV. This study was performed to investigate toxicity of GN01 in SD rats after weekly oral dosing for 28 days and to determine its no-observed-adverse-effect-level (NOAEL). Male and female SD rats were orally administered with GN01 at 0, 25, 50 and 100 mg/kg bw/day of dsRNA once per week for 28 days (total 5 administrations). The highest dose 100 mg/kg bw/day was determined based on the maximum volume injectable (20 mL/kg bw) via gavage. During treatment period, clinical signs, functional and sensory responses, body weights, food and water consumption, ophthalmological findings and urinalysis were investigated. After treatment period, hematological and clinical biochemistry tests and examination of necropsy findings, organ weights and histopathological lesions were performed. There were no significant differences between all test groups and vehicle control group in all measured parameters. Therefore, the NOAEL of GN01 was determined 100 mg/kg bw/day, the highest dose administered. In conclusion, repeated oral administration of GN01, a dsRNA medicinal product, is safe even at the maximum available dose in rats.

Ischemic stroke is caused by a blockage of the cerebral artery, which leads to a severe neurological disorder. Chlorogenic acid is a phenolic acid found mainly in plants such as coffee beans, eggplants, and carrots. It exerts a neuroprotective effect against cerebral ischemic damage. Bcl-2 family protein is a representative apoptosis regulatory protein. Bcl-2 and Bcl-xL act as apoptosis inhibitors, while Bax and Bad act as apoptosis inducers.The interaction of Bcl-2 family protein plays an important role in determining cell fate. The aim of this study was to investigate whether chlorogenic acid modulates the interaction of Bcl-2 family proteins during ischemic injury. Middle cerebral artery occlusion (MCAO) surgery was performed to induce cerebral ischemia. Chlorogenic acid (30 mg/kg) or phosphate buffered saline was intraperitoneally injected to adult male rats 2 h after MCAO surgery. Neurobehavioral tests were performed to confirm the neuroprotective effect of chlorogenic acid 24 h after MCAO injury, and immunoprecipitation analysis was performed to investigate the interaction of Bcl-2 family protein. MCAO damage showed signs of severe neurological disorders, while chlorogenic acid improved these disorders. Results of immunoprecipitation analysis were as follows. Interaction between Bax and Bcl-2 or Bcl-xL was decreased in MCAO injury, chlorogenic acid prevents these decreases. In contrast to Bax, Interaction between Bad and Bcl-2 or Bcl-xL was increased in MCAO injury, chlorogenic acid prevents these increases. Furthermore, chlorogenic acid attenuated MCAO-induced increase of capase-9. In conclusion, our findings demonstrate that chlorogenic acid exerts a neuroprotective effect against cerebral ischemic injury by modulating interaction of Bcl-2 family proteins.

The aim of this study was to investigate the hypoglycemic effects of saengshik in Sprague-Dawley (SD) rats and to explore the potential of three commercially available saengshik products (BS, LS, WS) as an alternative diabetic meal. Blood glucose levels were measured at 30, 60, 90, 120, and 150 minutes after the ingestion of experimental materials. In experiment 1, the amount of experimental materials remained the same. We measured blood glucose-related biomarkers as the area under the blood glucose response curve (AUC), glycemic index (GI), maximum concentration (Cmax), and time to reach maximum concentration (Tmax). AUC and Cmax of the experimental group showed significant differences compared to the control group, while GI and Tmax did not show significant differences among the groups but were lower in the experimental group compared to the control group. In experiment 2, carbohydrates were adjusted to the same amount. We measured blood glucose-related biomarkers in the same manner as Experiment 1 and obtained similar results. These hypoglycemic effects appear to be attributed to phytochemicals and dietary fiber found in whole, unrefined grains. These results suggest that saengshik exerts hypoglycemic effects by modulation of glucose-related biomarkers.

Hypertension caused by high-fat and high-salt diets is is a well-known significant risk factor for cardiovascular and cerebrovascular diseases. In this study, to confirm the relationship between hypertension and immune cells, angiotensin (Ang) II was administered to Dahl salt-sensitive (SS) rats and Dahl salt-resistant (SR) rats. Then the expression of immune cells and the proinflammatory cytokines were compared between the SS and SR rats. It was observed that after administration of Ang II (50ng/kg/min) for three weeks, blood pressure was increased in the SS rats, but there was no significant change in the SR rats. In addition, the expression of T helper (Th) cells and Th 17 cells in the spleen and the expression of Th cell Rorγt and regulatory T regulatory (Treg) cells in the peripheral blood mononuclear cells did not show a significant difference between the two experimental groups even after the administration of Ang II.IL-1β expression was significantly increased in the kidney tissue of the SS rats, while there was no significant difference in the IL-6 expression in all the experimental groups. The results of this study suggest that Ang II induces hypertension by stimulating IL-1β secretion from renal macrophage in SS rats.

This study examined the subacute oral toxicity of Dendropanax morbiferus H.Lév leaves hot-water extracts (DMWE) using male and female Spargue-Dawley rats. Rats were orally administered the DMWE at dose levels of 0, 250, 500, 1,000, and 2,000 mg/kg body weight (BW) for four weeks. For experimental period, clinical signs and the number of deaths were examined, and feed intake and BW of all experimental animals were measured once a week for four weeks. At the end of the experiment, blood samples were collected from all rats, and all animals were euthanized and autopsies were performed to collect major organs. No dead animals were found during the experimental period. In addition, no differences were found between control and DMWE-treated groups in feed intakes, BW changes, organ weights, clinical signs, hematological parameters, and serum biochemical parameters. The results of this study provided evidence that oral administration of DMWE at the dose of 2,000 mg/kg BW is safe in rats and may not exert severe toxic effects.

This study was designed to evaluate antihyperglycemic and antihyperlipidemic effects of ethanol extracts of Taraxacum mongolicum (T.m.) on streptozotocin (STZ)-induced diabetic rats. Sprague-Dawley rats were randomly assigned to five groups: normal (NC), STZ-control (DC), and three experimental groups. Diabetes was induced in Sprague-Dawley rats with a single intravenous injection [45 mg/kg body weight (b.w.)] of STZ. An ethanol extract of T.m. was orally given to diabetic rats for 14 days. Three experimental groups were additionally treated with T.m. extract at doses of 1 g/kg b.w./day for T.m.-1, 2 g/kg b.w./day for T.m.-2, and 3 g/kg b.w./day for T.m.-3. Oral administration of T.m.-2 significantly increased their body weights. T.m.-1 and T.m.-2 significantly decreased aspartate aminotransferase (AST) levels than DC. T.m.-1 and T.m.-2 group significantly decreased blood glucose levels. Total cholesterol, triglycerides, and free fatty acids were significantly decreased whereas high-density lipoprotein cholesterol was significantly increased in groups treated with T.m. extract than those in the DC group. These results support the fact that administration of T.m. extract can reduce hyperglycemia and hyperlipidemia risk in diabetic rats.

본 연구의 목적은 5% 녹두(Phaseolus aureus L.)의 급여가 고지혈증 유발 Sprague-Dawley (SD)계 흰쥐의 혈청 blood urea nitrogen(BUN), creatinine 및 요산(uric acid)의 농도와 aspartate aminotransferase(AST), alanine aminotransferase(ALT), alkaline phosphatase(ALP), lactate dehydrogenase(LDH), amylase, lipase 및 catalase 활성에 미치는 영향을 규명하고자 시행하였다. 연구 결과 녹두는 고지혈증으로 유발된 흰쥐의 혈청 BUN, creatinine, 요산 농도 및 AST, ALT, ALP, LDH, amylase, lipase의 활성 감소와 catalase 활성을 증가시키는 것으로 나타났다. 따라서 녹두는 신장과 간 등의 기능 개선과 예방에 효과적일 것으로 판단되어 기능성 소재로서의 이용 가능성이 있을 것으로 기 대된다.

The surfactant, 2-{2-[2-(4-nonylphenoxy)ethoxy]ethoxy}ethan-1-ol, is used in detergents, pesticides, cosmetics, and disinfectants. Since it is found in products that can be inhaled, we evaluated its toxicity to humans upon exposure. A total of 18 rats were exposed to nasal inhalation for 4 hr to determine the acute inhalation toxicity of 2-{2-[2-(4-nonylphenoxy) ethoxy]ethoxy}ethan-1-ol; the exposure concentrations were 5.0, 1.0, and 0.5 mg/L, with three males and three females at each concentration. After the end of the exposure, mortality, general symptoms, and weight changes were observed for 14 days, and autopsy findings were confirmed. The actual concentrations of the test substance in the chamber during the exposure were measured as 3.29, 1.03, and 0.52 mg/L, respectively. The delivered dose was 552.72, 173.04, and 87.36 mg/kg/day for males, and 829.08, 259.56, and 131.04 mg/kg/day for females. As a result of the test in the OECD Test Guideline 436, all animals exposed to a concentration of 3.29 mg/L died; three males and one female died out of six exposed to the 1.0 mg/L concentration. In addition, one died out of six males exposed to a 0.5 mg/L concentration. As a result, 2-{2-[2-(4-nonylphenoxy)ethoxy]ethoxy}ethan-1-ol was considered to be Globally Harmonized System of Classification and Labelling of Chemicals Category 2 (> 0.5–1 mg/L).

Iron deficiency is known to be a common nutritional disorder in many countries, especially among children, women of childbearing age and pregnant women. SUNACTIVE Fe-P80 is a new type of iron supplement that applies nanotechnlateology for the purpose of overcoming the disadvantages of food supplements. This study was conducted to investigate the potential adverse effects of a 28-day repeated oral dose of SUNACTIVE Fe-P80 in rats. SUNACTIVE Fe-P80 was administered once daily by gavage to Sprague-Dawley rats for 28 days at doses of 0, 500, 1,000, and 2,000 mg/kg/day. Additional recovery groups from the control and highdose groups were observed for a 14-day recovery period. At the scheduled termination, the animals were sacrificed, their organs weighed, and blood samples collected. There were no treatment-related effects in the context of clinical signs, body weight, food intake, ophthalmoscopy, urinalysis, necropsy findings, organ weights, and hematologic, serum biochemical and histopathological parameters at any dose tested. Under the present experimental conditions, the no-observed-adverse-effect level of SUNACTIVE Fe-P80 was ≥ 2,000 mg/kg/day in both the sexes, and no target organs were identified. Thus, the results suggest that SUNACTIVE Fe-P80 is relatively safe, as no treatment-related adverse effects were observed following a 28-day repeated oral dose experiment.

Aralia elata, Chaenomeles sinensis fruit, and Glycyrrhizae radix have been widely used as oriental medicinal plants in Korea, China and Japan and found to possess anti-oxidative and anti-inflammatory activities. The current study was conducted to investigate the neuroprotective effect of an ethanol extract of a mixture of A. elata, C. sinensis fruit, and Glycyrrhizae radix (ACG) against ischemia-induced brain injury in rats and excitotoxic and oxidative neuronal death in primarily cultured rat cortical neurons. Transient focal cerebral ischemia was induced by 2 h middle cerebral artery occlusion followed by 24 h reperfusion (MCAO/R) in rats. Oral administration of ACG (10, 25, and 50 mg/kg) 30 min before MCAO, after 1 h of MCAO, and after 1 h of reperfusion reduced MCAO/R-induced brain infarct and edema formation. ACG also inhibited development of behavioral disabilities in MCAO/R-treated rats. Exposure of cultured cortical neurons to 500 μM glutamate for 12 h resulted in neuronal cell death. ACG (1, 10, and 50 μg/mL) inhibited glutamate-induced neuronal death. Furthermore, ACG inhibited 100 μM hydrogen peroxide (H2O2)- and hypoxia-induced neuronal death. These results suggest that the neuroprotective effect of ACG against ischemia-induced brain damage might be associated with its anti-excitotoxic and anti-oxidative activity and that ACG may have a therapeutic role for prevention of neurodegeneration in stroke.

The rapid development of computer vision and deep learning has enabled these technologies to be applied to the automated classification and counting of microscope images, thereby relieving of some burden from pathologists in terms of performing tedious microscopic examination for analysis of a large number of slides for pathological lesions. Recently, the use of these digital methods has expanded into the field of medical image analysis. In this study, the Inception-v3 deep learning model was used for classification of chondrocytes from knee joints of rats. Knee joints were extracted, fixed in neutral buffered formalin, decalcified, processed and embedded in paraffin, and hematoxylin and eosin (H&E) stained. The H&E stained slides were converted into whole slide imaging (WSI), and the images were cropped to 79 × 79 pixels. The images were divided into training (60.42%) and test (39.58%) sets (46,349 and 30,360 images, respectively). Then, images containing chondrocytes were classified by Inception-v3 and accuracy was calculated. We visualized the images containing chondrocytes in WSIs by adding colored dots to patches. When images of chondrocytes in knee joints were evaluated, the accuracy was within the range of 91.20 ± 8.43%. Therefore, it is considered that the Inception-v3 deep learning model was able to distinguish chondrocytes from non-chondrocytes in knee joints of rats with a relatively high accuracy. The above results taken together confirmed that this deep learning model could classify the chondrocytes and this promising approach will provide pathologists a fast and accurate analysis of diverse tissue structures.

본 연구는 hesperidin과 hesperidin의 aglycone 형태인 hesperetin의 2,2'-azobis (2-aminopropane) dihydrochloride (AAPH)에 의해 신장 독성이 유도된 쥐에서 신장 보호 효과에 대해 연구하였다. Hesperidin과 hesperetin은 200 mg/kg/day의 농도로 7일간 위내투여하였으며, AAPH를 복강주사하여 급성 신장 손상을 유도하였다. 이 후 쥐의 신장 조직에서 지질과산화 함량, nitric oxide (NO) 생성량, catalase 효소 활성을 측정하였으 며, nuclear factor-kappa B (NF-κB) 및 inducible nitric oxide synthase (iNOS) 단백질 발현량을 측정하였다. AAPH로 신장 독성을 유도한 control군의 신장 내 지질과산화 및 NO 생성량은 신장 독성을 유도하지 않은 normal군에 비해 유의적으로 증가하여 산화적 손상이 유도됨을 확인하였다. 반면 hesperidin과 hesperetin를 투여했을 때 신장 내 지질과산화 및 NO 생성량이 control군에 비해 유의적으로 감소하여 산화적 스트레스 개선 효과를 확인하였다. Hesperidin과 hesperetin을 투여한 군의 경우 신장 내 항산화 효소인 catalase 활성을 유의적으로 증가시켰다. 뿐만 아니라, hesperidin과 hesperetin의 투여는 AAPH로 신장독성을 유도한 control군 100% 대비 NF-κB 단백질을 각각 66% 및 71%로, iNOS 단백질 발현을 각각 46% 및 33%로 억제시켰다. 따라서 본 연구를 통해 hesperidin과 hesperetin의 투여가 항산화 활성 조절을 통해 AAPH로 유도된 신장 독성을 억제하는 것을 알 수 있었다.

Declined salivary gland function is commonly observed in patients with diabetes. Advanced glycation end products (AGEs) are believed to contribute to the pathogenesis of diabetes-induces hypofunction of the salivary glands. Polydatin (resveratrol-3-O-β-mono-D-glucoside) is a polyphenol that can be easily accessed from peanut, grape, and red wines. Although polydatin is known to have anti-glycation, anti-oxidation, and anti-inflammation effects, its effect in the salivary gland is not known. In the present study, we evaluated the AGEs burden in the salivary gland in streptozotocin (STZ)-induced diabetic rats and the protective effect of polydatin on diabetes-related salivary hypofunction. Polydatin (50 and 100 mg/kg/day) was orally administered in the STZ-induced diabetic rats for 4 weeks. The results showed that the salivary flow rate of the STZ-induced diabetic rats was reduced compared with that in the normoglycemic control rats. The circulating AGEs in serum and secreted AGEs in saliva increased in the STZ-induced diabetic rats. The reactive oxygen specises (ROS) were highly generated in the salivary gland in these diabetic rats. The salivary gland from the diabetic rats showed increased acinar cell apoptosis compared to normoglycemic control mice. However, polydatin suppressed all of these diabetes-related salivary changes. Overall, polydatin could provide a beneficial option for diabetes-related salivary hypofunction.

The purpose of this study was to investigate the effects of extract mixture of C. wilfordii and P. umbrosa (IPLUS-CWPU) on bone growth in 4-week old young male SD rats. To confirm the effect of IPLUS-CWPU, we measured the length of bone growth plate, the ratio of proliferative zone to the length of growth plate and the expression level of insulin-like growth factor, IGF-1. The IPLUS-CWPU treatment shows a significant increase of tibial and femoral growth plate and the ratio of proliferative zone in growth plate. Especially, the length increased by 13.9% and 25.3% in the tibia and femur, respectively, in the high-dose group compared to the normal group. Moreover, the expression of IGF-1 gene in liver was upregulated in IPLUS-CWPU treated groups. These results indicated that IPLUS-CWPU administration could increase the proliferative zone of bone growth plate in early developmental stage by upregulation of IGF-1 gene.