Purpose : This study was peformed to evaluate the effect of Kamijoaguiem(JGE) on the bone mass and its related factors. Methods : We used ovariectomized rat as an estrogen-deficient animal model. The model rats of osteoporosis showed a significant decrease in bone density, bone ash density, calcium content of femur bone. At the 7th day after operating ovariectomy, rats were administered with JGE per orally, and continued for 10 weeks. And osteoporosis related parameters were determined to investigate the effect of JGE. Results : Bone density, bone ash density, bone calcium, magnesium and phosphorus was decreased in osteoporotic rats. JGE improved the decreased bone density, bone ash density and the decreased bone magnesium, but JGE didn't improve the decreased bone calcium and phosphorus in osteoporotic rats. Osteocalcin in serum and hydroxy-proline excretion in urine were increased in osteoporotic rats. Their levels were decreased when JGE was administered. ALP activity in serum was increased in osteoporotic rats. JGE didn't induce any significant changes. JGE showed significant increase in serum calcium level, total protein level, albumin level, BUN level, serum LDH activity. JGE didn't show significant increase in serum T-cholesterol density, triglyceride density, HDL-cholesterol density. JGE didn't show significant increase in RBC number, hemoglobin level, platelet number, hematocrit level. JGE showed inhibitory effect on the degradation of bone-matrix in osteoporotic rats, in histological examination to Hematoxylin-eosin stain. Conclusion : JGE might improve bone density due to inhibition of bone resolution in osteoporotic rats. It suggest that JGE may be useful prescription in osteoporosis.
Purpose : this study is to examine what are the effects of the Sintongchukeatang(SCT) on the ovariectomized rat model of postmenopausal osteoporosis. Methods : 12weeks Female Sprague-Dawley 30 rats of weighting 250-300g, were divided into three groups including the sham operation groups(10heads) and overiectomy groups(10heads). then we observed changes in the body weight serum metabolic products and femoral trabecular bone. Results : 1. The level of serum ALP activity in control group showed significant increase in comparison with sham, but that in SCT-treated was significantly decreased in comparison with control. 2. The level of serum GOT in control group showed no change in comparison with sham, but that in SCT-treated was significantly decreased in comparison with control. The level of serum GPT did not significant change among the three groups. 3. The level of serum estrogen in control group showed slightly decreased in comparison with sham, but that in SCT-treated showed no change in comparison with control. 4. Trabecular bone area as well as trabecular thickness in control group showed significant decrease in comparison with sham. Those in SCT-treated showed significant increase in comparison with control. 5. Trabecular separation only in SCT-treated showed significant decrease in comparison with control. 6. Osteoclast number and oseoblast surface in control group showed significant increase in comparison with sham. Those in SCT-treated showed significant decrease in comparison with control. Conclusion : SCT has shown to be capable of preventing and curing osteoporosis caused by old-aged and postmenopause.
Purpose : To address the ability of Dohaekseungkitang (DST: a commonly used herb formulation in Korea, Japan and China to have anti-cancer effect on cervical carcinoma), we investigated the effects of DST on programmed cell death (apoptosis) in HeLa human cervical carcinoma cells. Methods : We cultured HeLa cell which is human metrocarcinoma cell in D-MEM included 10% fetal bovine serum(Hyclone Laboratories) below $37^{\circ}C$, 5% CO2. Then we observed apoptosis of log phage cell which is changed cultivation liquid 24 Hours periodically. Results : After the treatment of DST for 48 hours, apoptosis occurred in a dose-dependent manner. In this study, we have shown that DST induces calpain and the associated caspase-8 and -9 activations. Apoptosis was prevented by pre-incubation of the cells with the calcium cHeLator-BAPTA-AM, calcium channel blocker-Nif edipine or Ryonidine agonist-Ryonidine peptide, implicating calcium in the apoptotic process. Ubiquitous calpains (mu- and m-calpain) have been repeatedly implicated in apoptosis, especially in calcium-related apoptosis. However this study showed 1hat either calpain inhibitor-calpastin or caspase-3 inhibitor-DEVD- did not blocked the herb formulation-induced apoptosis in HeLa human cervical carcinoma cells. D ST initiates a cell death pathway that is partially dependent of caspases. DST-induced apoptosis requires caspase-independent mechanism. Conclusion : We conclude that DST-induced calpain activation triggers the intrinsic apoptotic pathway in which caspase-independent mechanism is also involved.
Purpose : This study is to evaluate the synergistic cytotoxicity of Rhizoma Rehmanniae(RR), in adriamycin-treated HeLa human cervical carcinoma cells. Methods : We culture HeLa cell which is human metrocarcinoma cell in D-MEM included 10% fetal bovine serum(Hyclone Laboratories) below $37^{\circ}C$, 5% $CO_2$. Then we observed apoptosis of log phage cell which is changed cultivation liquid 24 Hours periodically. Results : The combination of RR and adriamycin synergistically augmented the cytotoxicity of HeLa cells. The apoptotic cell death was accompanied by the activation of caspase-3 and -8 as well as cleavage of poly(ADP- ribose) polymerase (PARP) in HeLa cells. The co-treatment of RR with adriamycin didn't have any effect on either the expression of Bcl-2 or that of Bax. Interestingly, a synergistic increase in apoptosis by the combination of two drugs was accompanied by the enhancement of Pas and Fas ligand (FasL) expression in HeLa cells. Taken together, the combination of RR and adriamycin significantly augmented the apoptotic cytotoxicity of Fas-positive cells, such as HeLa cells. The pathway is not involved in mitochondria-dependent pathway. Conclusion : RR induces apoptosis in HeLa cells via p38 MAPK activation.
Purpose : This study was to investigate the effects of Dohongsamul-tang extract on the rat with an surgically induced endometriosis. Methods : Four weeks after the operation for inducing the endometriosis to a matured female rat, the normal hyperplasia of the transplanted endometrium was identified in anatomico-histological aspects. By dividing each of the control group and Dohongsamul-tang administered experimental group into 8 rats, Dohongsamul-tang concentrates were administered orally to the experimental group with 1g/1ml/200g everyday for 40 days and then images were taken from the macroscopic tissues which were transplanted to the mesenterium, and the concentrations of progesterone, estradiol, tumor necrosis $factor-{\alpha}(TNF-{\alpha})$, and interleukin(IL)(-2, -4, -6, and -10) in the serum were measured. Results : Transplanted endometrium tissues showed the histological findings in accordance with the normal endometrium tissues and from the macroscopic findings the size of transplanted endometrium tissues showed a definite decrease. Compared to the control group, the experimental group showed significant decreases in the values of $TNF-{\alpha}$,IL-2, and IL-4 and significant increases in the values of IL-10. However, there were no significant differences in progesterone, estradiol and IL-6. Conclusion : From the above results, Dohongsamul-tang showed the strengthening of immunological function and anti-inflammatory effect. Therefore it is considered that Dohongsamul-tang will be very effective on the treatment of inflammation developed endometriosis, especially to treat the endometriosis without affecting the ovary functions.
Purpose : This study was conducted to investigate the effect of pregnancy and parity on bone marrow density using Dual Energy X-ray Absorptiometry (DXA) after parturition Methods : The observation cases who was over 20 and under 35 years old just after childbirth were admitted twice to woosuk university hospital from Aug 2000 to July 2005. During the first admission period, we measured the bone marrow density(BMD) using DXA in 13th day, and when the patient came to the hospital just after childbirth again, we followed up the BMD in 13th day. The evaluation index of this report was comparison of the T-score which was about the lumbar spine(L1-L4) BMD and femur neck BMD. Results : The continuous parturition was increased the lumbar spine BMD(P<<0.05), and decreased the femur neck BMD but it had no meaning. In the cases of the parturition interval under 24 months regarded as siblings born within a year of each other, the analysis results of BMD showed increase in lumbar spine BMD. In the cases of the parturition interval over 24 months, there was increased in lumbar BMD, and decreased in femur neck BMD. To the analysis of the weight variation, the increased BMI group has a significant increase in lumbar spine BMD, and the decreased BMI group also increase in lumbar spine BMD but there was no meaning about that. Conclusion : The continuous parturition was increased the lumbar spine BMD.
Purpose : This study was conducted to investigate the dose dependent effects of Panax Ginseng on the reproductive functions in mice. Methods : We used the 8-week-old mice, and administered 0.2ml extract solution of Panax Ginseng in the different concentration once a day for 60 days. The control group was administered 0.2ml normal saline in the same way and duration. We counted the total, motile and normal sperm number of the cauda epididymis and measured the activities of sperm hyaluronidase, peroxidase and catalase of the isolated testis tissues. And we observed histological changes of surgically isolated testis by histochemical methods. Results : All Panax Ginseng extract solution groups showed significantly dose dependent differences in the total number, the motility and normality of sperms compared with the control group, respectively. In the histological analysis of the testicular tissues, all Panax Ginseng extract solution groups showed the enlargement of testicular lobe diameter and apparent angiogenesis between seminiferous tubules. And the activity of typical sperm enzyme, hyaluronidase, was significantly increased in the Panax Ginseng extract solution groups compared to the control group. In the antioxidant activity analysis, the activities of peroxidase and catalase were significantly increased in the Panax Ginseng extract solution groups compared to the control group. Conclusion : This study shows that Panax Giseng has the beneficial effects on the concentration, morphology and motility of sperm, the activities of sperm hyaluronidase, testicular peroxidase and catalase. We can suggest that Panax Ginseng be useful for the treatment of male infertility.
Purpose : To address the ability of Myrrha (MY) to induce cell death, we investigated the effect of MY on apoptosis. In human cervical carcinoma HeLa cells, apoptosis occurred following MY exposure in a dose-dependent manner. Methods : We have tested several kinds of anti-oxidants to investigate the MY-induced apoptotic mechanism. Among the anti-oxidants, N-acetyl cysteine(NAC) or reduced glutathione (GSH) protects MY-induced apoptosis. NAC is an aminothiol and synthetic precursor of intracellular cysteine and GSH. To confirm the role of GSH in MY-induced apoptosis, methionine and cystathionine-glutathione extrusion inhibitors were treated in the presence of MY. Results : NAC, GSH, methionine or cystathionine led to protective effect against MY-induced apoptosis in HeLa cells. The GSH and GSH-associated reagents regulate MY-induced cytochrome c release and the resultant caspase-3 activation. Furthermore, the two specific inhibitors of carrier-mediated GSH extrusion, methionine and cystathionine demonstrate GSH extrusion occurs via a specific mechanism. While decreasing GSH extrusion and protecting against MY-induced apoptosis, methionine and cystathionine failed to exert anti-apoptotic activity in cells previously deprived of GSH. Conclusion : the target of the protection is indeed GSH extrusion. This shows that the protective effect is achieved by forcing GSH to stay within the cells during apoptogenic treatment. All this evidence indicates the extrusion of GSH precedes andis responsible for the apoptosis, probably by altering the intracellular redox state, thus giving a rationale for the development of redox-dependent apoptosis in MY-treated human cervical carcinoma HeLa cells.
Purpose : This study was aimed to investigate the inhibitory effect of Corydalis Tuber on the proliferation of human uterine leiomyoma cell and the expression of gene related the mechanism of cell apoptosis. Methods : We counted the number of suvival cells treated with indicated concentration of Corydalis Tuber and investigated cell viability by MTS assay. Furthermore, flow cytometric analyis were used to dissect between necrosis and apoptosis related with cell cycle and then we observed the differential gene expression by western blot analysis. Results : 1) The inhibitory effect on the proliferation of uterine leiomyoma cell treated with Corydalis Tuber was increased in a concentration and time proportional. 2) The result of flow cytometry analysis, subG1 phase arrest related cell apoptosis was not investigated in uterine leiomyoma cell treated Corydalis Tuber but showed G2/M phase prolongation. 3) The gene expression of p27, p21 related cell cycle was increased according to increasing concentration, but p53 was not exchanged. 4) The dephosphorylation of pRb gene were increased dependent on treatment concentration and pro-caspase 3, CDK4 were not exchanged. Conclusion : This study showed that Corydalis Tuber have the inhibitory effect on the proliferation of human uterine leiomyoma cell but the effect was thoughted no relationship with apoptosis. The inhibitory effect was suggested that dephosphorylation of pRb gene induced with increasing p21, p27 prolonged cell division in G2/M phase.
Purpose : This study was aimed to investigate the inhibitory effect of Sparganii Rhizoma on the proliferation of human uterine leiomyoma cells and the expression of gene related the mechanism of cell apoptosis. Methods : This study was evaluated the number of death cells treated with indicated concentration of Sparganii Rhizoma and investigated cell death rate by MTS assay. Furthermore, fluorescence-activated cell sorter analysis and DNA fragmentation assay were used to dissect between necrosis and apoptosis. and then we observed the differential gene expression by western blot analysis. Results :1) The inhibitory effect on the growth of uterine leiomyoma cell treated with Sparganii Rhizoma was increased in a dose dependent manner. 2) As the result of FACS analysis, subG1 phase incrase was observed 23.49% inuterine leiomyoma cell treated with Sparganii Rhizoma at $500\;{\mu}g/ml$ compared to control.. 3) The gene expression of p53, p21 related cell apoptosis was increased according to increasing concentration but p27 was none exchanged. 4) The expression of cyclin A, D and E was decreased in a concentration proportional and then the dephosphorylation of pRb was increased. 5) The character of apoptosis, DNA fragmentation was significantly observed according to increasing concentration. 6) The expression of pro-caspase3 were decreased dependent on treatment concentration and activated PARP took place. Conclusion : The inhibitory effect of Sparganii Rhizoma on the proliferation of human uterine leiomyoma cells was observed with apoptosis and cell cycle arrest. These data suggest that Sparganii Rhizoma might be candidate of medical therapy for uterine leiomyoma.
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