• The Journal of Internal Korean Medicine
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• v.33 no.3
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• pp.272-283
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• 2012

Objectives : The purpose of this study was to investigate the mechanism of protective effect of Jinmu-tang (JMT, Zhenwu-tang) extract on $H_2O_2$-induced cell death in C6 glial cells. Methods : Cultured C6 glial cells of white mice were pretreated with JMT extract and exposed to $H_2O_2$ for inducing cell death. We measure the cell viability by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) assay and investigate the cell morphology using a light microscope after crystal violet (CV) staining. Reactive oxygen species (ROS) formation was analyzed using a flow cytometer and a fluorescent microscope after staining with 2'7'-dichlorofluorescein diacetate (DCF-DA). DNA fragmentation was analyzed using a flow cytometer after propidium iodide (PI) staining and nuclei morphology was investigated using a fluorescent microscope after 2-[4-amidinophenyl]-6-indo-lecarbamidine dihydrochloride (DAPI) staining. We analyzed expression of Bax, processing of procaspase-3 and poly (ADP-ribose) polymerase (PARP), and activation of nuclear factor-${\kappa}B$ (NF-${\kappa}B$) by western blot method. Tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$) secretion was analyzed using Quantikine kit. Results : We determined the elevated cell viability by JMT extract on $H_2O_2$-induced C6 glial cell death. ROS formation, DNA fragmentation, $I{\kappa}B{\alpha}$ phosphorylation, NF-${\kappa}B$ activation, and secretion of TNF-${\alpha}$ induced by $H_2O_2$ are inhibited by JMT extract pre-treatment. JMT extract inhibits Bax expression, processing of caspase-3 and PARP that are critical biochemical markers of apoptotic cell death. Conclusions : These results suggest that JMT extract has a protective effect on $H_2O_2$-induced C6 glial cell death in various pathways.

• Journal of Korean Medicine Rehabilitation
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• v.30 no.2
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• pp.19-35
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• 2020

Objectives The aim of this study is to evaluate the fracture healing effect of Jinmu-tang (JM) on femur fractured rats. Methods Rats were randomly divided into 5 groups (normal, control, positive control, JM extract with low concentration and JM extract with high concentration). All group except normal group went through both femur fracture. Normal and control group received no treatment at all. Positive control group were medicated with tramadol (20 mg/kg) once a day for 14 days. Experimental group was orally medicated with JM extract (10 mg/kg for low concentration, 50 mg/kg for high concentration) once a day for 14 days. In order to investigate fracture healing process, plasma and serum were obtained. Also, micro-computed tomography was conducted to see the frature site visually. Immunohistochemistry for transforming growth factor-β1, Ki67, alkaline phosphatase, runt-related transcription factor 2, receptor activator of nuclear factor kappa-β, tartrate resistant acid phosphatase was conducted to observe bone healing progress after 14 days since fracture occured. Aspartate aminotransferase, alanine aminotransferase, blood urea nitrogen and creatinine levels were measured in plasma, for hepatotoxicity and nephrotoxicity of JM extract. Osteocalcin was measured to observe activity of osteoblast. Results Through Micro-CT, more fracture healing was observed on both experimental group than control and positive control group. Through Hematoxylin & Eosin and safranin O staining showed bone cell proliferation and bone formation in the experimental group. RANK was significantly increased in the experimental groups. JM with high concentration showed statistically significant of TGF-β and Osteocalcin. NO, TRAP and ALP were not significantly changed. Liver toxicity was not significantly observed. Creatinine significantly increased in both experimental groups after 28 days. Conclusions As described above, JM extract showed anti-inflammatory effect, promoted fracture healing by stimulating the bone regeneration factor, and showed little hepatotoxicity and nephrotoxicity. In conclusion, JM extract can promote fracture healing and it can be used clinically to patients with fracture.