Kim, Chung-Hui;Han, Jin;Kim, Na-Ri;Park, Ju-Hee;Yang, Young-Churl;Kim, Eui-Yong
The Korean Journal of Physiology and Pharmacology
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v.5
no.3
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pp.223-230
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2001
Melatonin, a pineal gland hormone, is believed to act as an antioxidant via the stimulation of radical detoxifying enzymes and scavenging of free radicals. In this study, effects of in vitro and in vivo treatments of melatonin on the cisplatin-induced lipid peroxidation, LDH release and plasma creatinine were determined in rabbit renal cortical cells. The level of malondialdehyde (MDA) was assayed as an index of lipid peroxidation and the level of LDH release as an indicator of cellular damage. In in vitro studies, cisplatin increased the levels of MDA and LDH release in a concentration-and time-dependent manner. Melatonin inhibited the cisplatin-induced lipid peroxidation and LDH release in a concentration-dependent manner. The minimal effective concentration of melatonin that significantly reduced the $300\;{\mu}M$ cisplatin-induced lipid peroxidation and LDH release was 1 mM. In in vivo studies, the levels of lipid peroxidation and LDH release in renal cortical cells increased significantly 24 or 48 hours after a single injection of cisplatin (6 mg/kg). When the cisplatin-injected rabbits were pretreated with 10 mg/kg of melatonin, a significant reduction in both lipid peroxidation and LDH release was observed. The plasma creatinine level increased from $0.87{\pm}0.07$ mg/dl in control to $6.33{\pm}0.54$ mg/dl in cisplatin-injected rabbits (P<0.05). Melatonin partially prevented the increase in serum creatinine level $(1.98{\pm}0.11\;mg/dl)$ by cisplatin (P<0.05). In the proximal tubules from cisplatin-treated group, tubular cells had microvilli of variable heights. Necrotic debris was seen in tubular lumens. In most of cells, the mitochondria and lysosomes were increased in frequency. The endocytic vacuoles were not prominent and distribution of the brush border was irregular and shortened. These cisplatin-induced morphological changes were moderate in the melatonin-pretreated group. These results suggest that the toxicity of cisplatin is associated with the generation of reactive oxygen free radicals and that melatonin is a powerful antioxidant, which prevents some of the adverse effects of cisplatin.
After Vanherweghem J-L reported the rapidly progressive interstitial renal fibrosis that developed in patients taking the slimming preparation compounded with chinese Herb in Belgium 1993. Chinese Herb Nephropathy(CHN) has become known as a new renal disease, CHN is described as the decrease of urinary renal enzyme, neutral endopeptidase(NEP). N-acetyl-${\beta}$-Dglucosaminidase(NAG). increase of urinary low molecular protein, ${\alpha}$1-microglobulin, ${\beta}$2-microglobulin. clara cell protein(CC16), retinol-binding protein(RBP) in clinical findings, and the proximal tubular atrophy, interstitial fibrosis. urothelial atrophy. glomerular sclerosis in histology, Because CHN was caused by Chinese herb contained in slimming preparation, western medical doctors have thought that all Herb medicine might have caused renal disease and prohibit the taking of any Herb medicine, However. CHN was actually caused by the aristolochic acid contained in some Herb medicines. Aristolochia manshuriensis, Aristolochia fang chi, which is the substitutions of Akebia quinata, Stephania tetrandra has being used in clinical. Aristolochia manshuriensis. Aristolochia fang chi were different with Akebia quinata. Stephania tetrandra in botany, and it have not been classified with medicines in Oriental medicine, That is, the aristolochic acid, not Herb medicines. causes CHN, So, Chinese Herb Nephropathy should be changed to Aristolochic acid Nephropathy.
Morphological features of the interaction between the hatching blastocyst and implantation in pig were studied by electron microscopy. The observations extended from late blastocyst stage to the completion of trophoblastic erosion of the epithelium and early decidual transformation of the epithelium and early decidual transformation of the stromal cells. Between day 7 and 17 of pregnancy, blastocysts from 0.3 to 12 mm in diameter were flushed from the uterine horns of Dutch Landrace pigs. On the 7th of development in the pig blastocyst, the blastocyst shedded of the zona pellucida established the tips of microvilli and with bleb-like cytoplasmic protrusions of the epithelial cells. From day 11 on in pig embryo, the bilayered trophoblast undergoes a dramatic phase of elongation so that the initially spherical expanded blastocyst becomes tubular. In pig, close apposition to the uterine wall beg-ins at about 12 $^1$/$_2$ days and then attachment occurred during the afternoon of the 16th or 18th day post coitum. At this stage, embryonic loss compared with corpus luteum number is up to 40% of ovulated oocytes. Therefore, the implantation failture of these embryos may be mainly caused by morphological abnormality and failture of zona shedding.
For the purpose of stydying the pharmacodynamic action of methemoglobin, the author made the following experiments: 1. Preparation of hemoglobin and methemoglobin solutions: Red cell suspension from rabbit blood was hemolysed with distilled water and then divided into two portions. One portion was dialysed through cellophane paper and made isotonic with the proper amount of sodium chloride. The second portion was treated with sodium nitrite to convert hemoglobin to methemoglobin, dialysed through cellophane paper and made isotonic. 2. The concentration of methemoglobin in solution, plasma and urine was determined by Horecker and Brackette's method, and that of hemoglobin by the cyanmethemoglobin method. 3. The concentration of methemoglobin and hemoglobin in the plasma and urine of rabbits was measured at several intervals of time after infusion of the above samples. 4. The blood pressure and respiration of rabbits were recorded on a kymograph, and the effects of the samples on them were observed. 5. The effects of the samples on the movements of the in-situ heart and the isolated intestine of rabbits were studied. 6. The kidneys of rabbits were excised 4 to 5 hours after injection of the samples, and histopathological examinations were made. These experiments revealed the following results: 1. When methemoglobin solution was allowed to stand in room air, there was no decrease in the concentration of methemoglobin. 2. When methemoglobin solution was mixed with whole blood and incubated at $37^{\circ}C$, the concentration of methemoglobin decteased gradually. 3. After the infusion of methemoglobin and hemoglobin solutions, the rate of disappearance of methemoglobin in the plasma was more rapid than that of hemoglobin in the plasma. The higher the initial concentration in the plasma, the larger was the rate of disappearance of methemoglobin. 4. The rate of disappearance of methemoglobin was exceedingly rapid for 30 minutes after the infusion. 5. The urinary excretion of methemoglobin was more rapid than that of hemoglobin. 6. It would seem that the circulating blood contains substances which are promptly mobilized in the plasma to reduce methemoglobin to hemoglobin. 7. Moderate amounts of methemoglobin solution caused some rise in the blood pressure and a transient acceleration of the respiration of the rabbits. These effects of methemoglobin were milder than those of hemoglobin. 8. The movements of the in-situ heart and the isolated intestine of rabbits were accelerated by methemoglobin. These accelerating effects were milder than those of hemoglobin. 9. In the kidneys of rabbits treated with methemoglobin solution, hyperemia of the glomeruli, cloudy swelling and hemoglobin deposit in the tubular epithelium, hemoglobin casts in the tubular lumina of the proximal tubules, and interstitial congestion were constantly observed. There was no definite difference between the histological findings in the rabbit kidneys injected with methemoglobin, and those injected with hemoglobin solutions.
Park, Sang-Joon;Lee, Sae-Bom;Lee, Young-Ho;Ryu, Si-Yun;Jeong, Kyu-Shik;Lee, Cha-Soo
Korean Journal of Veterinary Pathology
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v.3
no.1
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pp.15-25
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1999
To elucidate the mechanism of age-related development in FGS/NgaKIST mice with spontaneous glomerulosclerotic lesion, we examined expression and localization of various cytokine mRNA in the kidney in the progression of diseases. This mouse model is the first to develop spontanously occuring glomerosclerotic lesion in the kidney. In this study, we detected the up-regulation of local cytokine genes such as IL-1$\beta$, IL-2, IL-6, IL-10, TNF-$\alpha$, TGF-$\beta$, and IFN- $\gamma$ in the kidneys. In RT-PCR and Southern blot analysis, we detected gradual expressions of cytokine mRNA of IL-1$\beta$, IL-2, IL-6, IFN- $\gamma$, and TNF $\alpha$ mRNA during the course of disease. Other cytokines including IL -10 and TGF -$\beta$ were found to be appeared the slightly expressed level at 3 to 12 weeks before onset of inflammatory lesion but they are highly expressed at the end-stage of the disease accompaning high proteinurea and wasting. In situ RT-PCR, each cytokine mRNA were specifically localized in a variety of cells including mesangial, endothelial, parietal epithelial, tubular epithelial, arterial muscle cell, and infiltrated inflammatory cells. In addition, TNF - $\alpha$was detected moderately in the visceral and parietal epithelial cell, but weakly in endothelial and mesangial cells, whereas IL-1 $\beta$ and IL -6 were strong in mesangial regions. IL-6 and TNF- $\alpha$ was highly localized in the damaged proximal and collecting tubules. Especially, TGF -$\beta$ mRNA was highly found in mesangial cells within glomerulus and interstitium during the end-stage of this disease.. These results indicate that pro inflammatory cytokines such as IL-1 $\beta$, IL-2, IL-6, and TNF- $\alpha$ were gradually expressed from the early stage of this disease to the end-stage, and that IL-10 and TGF-$\beta$ may be important in the accumulation of extracellular matrix(ECM) within glomerulus and periglomerular fibrosis in the progression of this disease as well as tissue destruction in end-stage of this disease.
Fine structures of retina of an ommatidium in dragonfly, having eyes of closed rhabdom type, were studied under light and electron microscopes. The ommatidia consisted of eight retinular cells distributed in a circular pattern and the retinular layer in turn can be divided into three sublayers according to the number of cells in the retina. Each retinular cell has different starting points in the retina and the length of retinular cells is varied greatly; the length of one distal retinular cell shows one half of that of others. In the middle layer, three proximal retinular cells interconnect the adjacent two rhabdoms which are triangular in the appearence of the cross section which in turn consisted of tubular, parallel and lamellated microvilli. The rhabdom is formed by three rhabdomeres, each of which is separated by $120^{\circ}$ between them, but they can be distinguished into two parts according to electron density. Around the outer part of microvilli composing rhabdom, electron density was much less than the inner part of the structure. The microvilli of the inner part appear to be connected to the cytoplasm of retinular cells. Rough endoplsmic reticulum with enlarged cisternae runs through the vacuoles in the outer part of distal retinular cells. Abundant mitochondria concentrated in the vicinity of rhabdom are found at the central part of the retinular cells, while in the area of immediate vicinity of the rhabdom, prominent vacuoles are observed. Above the rhabdom of an ommtidium stands a crystalline cone which is consisted of four cone cells arranged radially along the axis. The crystalline cone is surrounded by cells containing pigment granules. The outermost photoreceptor element of an ommatidium is corneal lens.
Ultrastructure of germ cells, the cyst epithelial cells and interstitial cells during spermatogenesis of the stone flounder, Kareius bicoloratus (Pleuronectidae) sampled on the west coast of Korea were investigated by electron microscopic observations. In the primary spermatocyte, the synaptonemal complexes appear in the zygotene stage of the prophase during maturation division. In the growing testis, especially, the interstitial cells (Leydig cells) appear near the primary, secondary spermatocytes and spermatids. Well-developed interstitial cells (steroid hormone secreting cells) which are located in the interlobular space in growing testis have three morphological characteristics of a vesicular nucleus, mitochondria with tubular cristae and smooth endoplasmic reticulum. During spermatogenesis, the primary and secondary spermatocytes attach to the cyst epithelial cell (Sertoli cell) having an elongated ovoid or triangular nucleus and several mitochondria in the cytoplasm. In the growing testis, lipid droplets, the mitochondrial rosettes and glycogen particles appear in the cytoplasm of the cyst epithelial cells near the secondary spermatocytes and spermatids. Particularly, the mitochondria, endoplasmic reticulum, little lipid droplets and the large amount of glycogen particles are present in the cytoplasm of the cyst epithelial cell in the late growing testis. In the late stage of spermiogenesis, the proximal centriole is joined to the nuclear envelope, the distal centriole forms the basal body of the flagellum and gives rise to the axial filament of the flagellum. No acrosome of the sperm is formed as seen in other teleost fish. The head of the spermatozoon is approximately $3{\mu}m$ in length and its tail is about $30{\mu}m$ in length. The axoneme of the tail flagellum of the spermatozoon consists of nine outer doublet microtubules at the periphery and two centrial singlet microtubules at the center. The spermatozoon of this species has two axonemal lateral fins. Especially, the cyst epithelial cells which located near groups of gametes in the various stages, show three functions: nutrition, phagocytosis and steroidogenesis. Especially, the nuclei of cyst epithelial cells in the recovery stage of the testicular developmental stages appear to be irregular in shape after spermiation. Of three functions of the cyst epithelial cell, several characteristics of phagocytosis are showed in the cytoplasm of the cyst epithelial cells in the recovery stage of the testicular developmental stages. At this stage, therefore, it is assumed that the cyst epithelial cells are involved in degeneration and resorption of undischarged germ cells after spermiation.
Juglans sinensis Dode has been reported to have antioxidant activity. However, the effect of Juglans sinensis Dode aquacupuncture(JS) on reactive oxygen species(ROS)-induced alterations in membrane transport function in renal tubular cells. This study was performed to evaluate the effect of JS on the organic hydroperoxide t-butylhydroperoxide(tBHP)-induced inhibition of $Na^+$-dependent phosphate($Na^+$-Pi) uptake in opossum kidney (OK) cells, an established renal proximal epithelial cell line. tBHP inhibited $Na^+$-Pi uptake in a time-dependent manner. The inhibitory effect of tBHP was prevented by JS over concentration range of 0.05-1mg/100ml in a dose-dependent manner. Kinetic studies showed that tBHP caused an decrease in Vmax for $Na^+$-Pi uptake without any a significant change in Km. $Na^+$-dependent phosphonoformic acid binding, a irreversible inhibitor of renal $Na^+$-Pi uptake, was decreased by tBHP treatment. The reduction in Vmax and phosphonoformic acid binding by tBHP was prevented by JS. tBHP induced lipid peroxidation and its effect was completely inhibited by JS and antioxidant N,N'-diphenyl-p-phenylenediamine. These data suggest that the oxidant inhibits phosphate uptake by a reduction in the number of active carrier across the membrane. JS may prevent oxidant-induced inhibition of membrane transport function by a mechanism similar to antioxidants in renal epithelial cells. Although the precise constituents remain to be explored, JS may be employed as a useful candidate herb for drug development to prevent and treat oxidant-mediated renal failure.
First, mice were dosed with 50mg/kg of streptozotocin(STZ) twice every 24 hours to cause high blood-sugar. Then, after three days, mice were injected with 100mg/kg of STZ again. Two different dosages of Saengjihwangeumja-gami were given to the experiment groups: SA group, 15mg/kg/day, and SB group, 90mg/kg/day, in order to determine the effects of Saengjihwangeumja-gami, which has been known to be good for DM(Diabetes Mellitus). By observing weight and blood-sugar level changes, blood tolerance, the numerical value of BUN(Blood Urea Nitrogen) and creatinine in blood, and through light-electronicmicroscopic and immunohistologic investigations of pancreas and kidneys, the following results were obtained: 1. The experiment groups showed a high suppressive effect of weight-loss. 2. The experiment groups' blood-sugar and blood tolerance showed an effective lowering of blood-sugar levels. 3. The experiment groups did not show any noticeable change in the numerical value of BUN and creatinine in blood compared with that of the control groups. 4. The experiment groups showed a higher Insulin positive reaction of pancreatic islets ${/beta}-cell$ than the control groups. 5. The experiment groups showed a higher immuno-reaction against IGF- II than the control groups. 6. Observation of apoptosis of the pancreatic islets showed that the cells of experiment groups were less injured compared with those of the control groups, and fewer apoptag-positive reaction cells were seen in experiment groups than in the control groups. 7. Uunder electron-microscopy, the insulin-containing granules in pancreatic islets ${/beta}-cells$ had increased more in the experiment groups than in the control groups. 8. Under light microscopy, the injury on the inner & outer membrane of the glomerulus and epithelial cells of capillaries and cells among vessels were fewer in the experiment groups than in the control groups. 9. More apoptag-positive reaction cells in the kidney were seen in the control groups than in the experiment groups. 10. PAS-positive reaction substances had increased more in the substrate among the vessels of a glomerulus belonging to the control group than those of the experiment group. 11. Uunder electron-microscopy, the nucleonic membrane, nucleoplasm and mitochondria of proximal and distal renal tubular were more injured in the control groups than in the experiment groups. In conclusion, strong evidence for the efficacy of Saengjihwangeumja-gami in lowering blood-sugar, and in recovery and generation of pancreatic tissues injured by DM was observed. Results suggest Saengjihwangeumja-gami is an effective treatment for DM. Further study of the principles of blood-sugar dropping effects of Saengjihwangeumja-gami are needed, as well as further study of recovery and regeneration of pancreatic tissues injured by DM.
Morphological difference of the renal glomerulus at different age groups have been studies in one week-old, five weeks-old, eight weeks-old, six months-old, twelve months-old, eighteen months-old, twenty-four months-old, and thirty months-old ICR mice. Pieces of the tissue taken from the renal corticies were fixed in 2.5% glutaraldehyde-1.5% paraformaldehyde solution (0.1 Millonig's phosphate buffer pH 7.3), and 1% osmium tetroxide solution (0.1 M Millonig's phosphate buffer, pH 7.3), and were embedded in araldite mixture. The ultrathin sections were stained with uranyl acetate and lead citrate solution, and were observed under a JEM 100CX-II electron microscope. The results were as follows: 1 In the one week-old mouse, thicknesses of the three layers of the glomeluar basal lamina (lamina densa, lamina rara interna and lamina rara externa) are similar, but in the five weeks-old mouse, thick lamina densa becomes a greater portion of the thickness of whole glomerular basal lamina. 2. No difference was noticed between thickness of the renal glomerular basal lamina of the five weeks-old mouse compare with that of the one week-old one, but basal lamina of the eight weeks-old one is thickened considerably and thicknesses were maintained through twelve months-old one. After eighteen months, the thickness of the glomerular basal lamina is increased remarkably. 3. After eighteen months, electron dense deposits within the basal lamina of the renal glomeruli are observed frequently. 4. Amount of the microfilaments in the mesangial cells and the mesangial matrices are increasing during aging. 5. The thicknesses of the basal laminae of the Bowman's capsule are increasing during aging. 6. After twenty four months, the proximal tubular cell-like parietal cells with well developed microvilli are observed frequently. From the above results, it was suggested that the renal glomerulus matures structurally in five weeks, and the function of the glomerulus is suppressed after eighteen months.
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