Dynamic changes of capillarization and peri-sinusoid fibrosis in alcoholic liver diseases.

AIM: To investigate the dynamic changes of capillarization and peri-sinusoid fibrosis in an alcoholic liver disease model induced by a new method. METHODS: Male SD rats were randomly divided into 6 groups, namely normal, 4 d, 2 w, 4 w, 9 w and 11 w groups. The animals were fed with a mixture of alcohol for designated days and then decollated, and their livers were harvested to examine the pathological changes of hepatocytes, hepatic stellate cells, sinusoidal endothelial cells, sinusoid, peri-sinusoid. The generation of three kinds of extra cellular matrix was also observed. RESULTS: The injury of hepatocytes became severer as modeling going on. Under electronic microscope, fatty vesicles and swollen mitochondria in hepatocytes, activated hepatic stellate cells with fibrils could been seen near or around it. Fenestrae of sinusoidal endothelial cells were decreased or disappeared, sinusoidal basement was formed. Under light microscopy typical peri-sinusoid fibrosis, gridding-like fibrosis, broaden portal areas, hepatocyte's fatty and balloon denaturation, iron sediment, dot necrosis, congregated lymphatic cells and leukocytes were observed. Type I collagen showed an increasing trend as modeling going on, slightly recovered when modeling stopped for 2 weeks. Meanwhile, type IV collagen decreased rapidly when modeling began and recovered after modeling stopped for 2 weeks. Laminin increased as soon as modeling began and did not recover when modeling stopped for 2 weeks. CONCLUSION: The pathological changes of the model were similar to that of human ALD, but mild in degree. It had typical peri-sinusoid fibrosis, however, capillarization seemed to be instable. It may be related with the reduction of type IV collagen in the basement of sinusoid during modeling.

[Effect of electroacupuncture intervention on autophagy pathway in APP 695 V 717 I transgenic mice].

OBJECTIVE: To observe the effect of electroacupuncture (EA) therapy on the intraneuronal Abeta1-42 and dysfunction of autophagy pathway, so as to reveal its mechanism underlying improvement of Alzheimer's disease (AD). METHODS: APP 695 V 717 I transgenic female mice were randomly divided into model group (n = 6) and EA group (n = 6); and C 57 BL/6 mice were used as the control group (n = 6). After 3 months' treatment by EA therapy at "Baihui" (GV 20) and "Yongquan" (KI 1) (15 min, once every other day, 2 Hz/100 Hz, 1-2 mA), the expression level of Abeta1-42 of the striate cortex was detected by immunohistochemistry. TUNEL staining was used to detect the degree of apoptosis of the striate cortex, and ultrastructural changes of autophagosome in the cortex were observed using electron microscope. RESULTS: In comparison with the control group, Abeta1-42 expression level and the apoptotic neurons in the striate cortex were significantly up-regulated in the model group (P < 0.01). Following EA intervention for 3 months, the Abeta1-42 expression level and the number of apoptotic neurons were significantly decreased in the EA group (P < 0.01, P < 0.05). Accordingly, transgenic induced dark degenerated neurons exhibiting irregular body deformation, analosis, and abundant secondary lysosomes and autophagosomes were reduced in the EA group. CONCLUSION: EA intervention can effectively down-regulate Abeta1-42 expression and number of the apoptotic neurons in the striate cortex in APP transgenic model mice, which may contribute to its effect in improving pathological changes of ultrastructure of neurons.

[Effect of moxibustion on hepatic glycogen and ultrastructure of exercise-induced fatigue rats].

OBJECTIVE: To observe the effect of moxibustion of "Zusanli" (ST 36) on hepatic glycogen-level and ultrastructure changes in exercise-induced fatigue rats. METHODS: Thirty-three SD rats were randomly and equally divided into control, model and moxibustion groups. The fatigue model was established by forcing the rat to have a loaded exhaustion swim, once daily for 21 days. Moxibustion was applied to bilateral "Zusanli" for about 30 min, once every other day for 11 times. The hepatic glycogen content was detected by chromatometry and the hepatic ultrastructure was observed by using transmission electron microscope. RESULTS: The hepatic glycogen content in the fatigue model group decreased significantly compared with the control group(P<0. 01), and that in the moxibustion group was increased significantly compared with the model group(P<0. 05). Under transmission electron microscope , it was found that the glycogen in the hypatocytes of the model was decreased and the ultrastructure of mitochondria and rough endoplasmic reticulum was unclear in outline. In moxibustion group, more glycogen granules were found in hepatocytes, and the structure of mitochondria and rough endoplasmic reticulum was basically clear. CONCLUSION: Moxibustion may increase hepatic glycogen content and improve ultrastructure of hypatocytes in fatigue rats, which may be part of its mechanism underlying relieving exercise-induced fatigue.

Neuroprotective effects of salvianolic acid B against oxygen-glucose deprivation/reperfusion damage in primary rat cortical neurons.

BACKGROUND: Cerebral ischemia-reperfusion injury is the main reason for the loss of neurons in the ischemic cerebrovascular disease. Therefore, to deeply understand its pathogenesis and find a new target is the key issue to be solved. This research aimed to investigate the neuroprotective effects of salvianolic acid B (SalB) against oxygen-glucose deprivation/reperfusion (OGD/RP) damage in primary rat cortical neurons. METHODS: The primary cultures of neonatal Wister rats were randomly divided into the control group, the OGD/RP group and the SalB-treatment group (10 mg/L). The cell model was established by depriving of oxygen and glucose for 3 hours and reperfusion for 3 hours and 24 hours, respectively. The neuron viability was determined by MTT assay. The level of cellular reactive oxygen species (ROS) was detected by fluorescent labeling method and spin trapping technique respectively. The activities of neuronal Mn-superoxide dismutase (Mn-SOD), catalase (CAT) and glutathione peroxidase (GSH-PX) were assayed by chromatometry. The mitochondria membrane potential (ΔΨ(m)) was quantitatively analyzed by flow cytometry. The release rate of cytochrome c was detected by Western blotting. The neuronal ultrastructure was observed by transmission electron microscopy. Statistical significance was evaluated by analysis of variance (ANOVA) followed by Student-Newman-Keuls test. RESULTS: OGD/RP increased the level of cellular ROS, but decreased the cell viability and the activities of Mn-SOD, CAT and GSH-PX; SalB treatment significantly reduced the level of ROS (P < 0.05); and enhanced the cell viability (P < 0.05) and the activities of these antioxidases (P < 0.05). Additionally, OGD/RP induced the fluorescence value of ΔΨ(m) to diminish and the release rate of cytochrome c to rise notably; SalB markedly elevated the level of ΔΨ(m) (P < 0.01) and depressed the release rate of cytochrome c (P < 0.05); it also ameliorated the neuronal morphological injury. CONCLUSION: The neuroprotection of SalB may be attributed to the elimination of ROS and the inhibition of apoptosis.

[Effect of electroacupuncture on the behavior and hippocampal ultrastructure in APP 695 V 717 I transgenic mice].

OBJECTIVE: To investigate the ultrastructural basis underlying electroacupuncture (EA) induced improvement of Alzheimer disease (AD) in transgenic mice. METHODS: Twelve APP 695 V 717 I transgenic mice were randomly divided into model group and EA group; and other 6 negative transgenic mice (C 57 BL/6 J) were made up of normal control group. After 3 months treatment by EA (15 min per other day, 2 Hz/100 Hz, 3-4 mA) applied to "Baihui" (GV 20) and "Yongquan" (KI 1), the learning and memory ability of mice was measured by Lashley III water maze test, and the ultrastructural changes of hippocampal CA 1 region was observed by electronic microscopy. RESULTS: The swimming escape latency and the number of navigating errors (dead-end forward swimming) in model group were significantly longer and more than those in normal control group (P < 0.05); and those in EA group were considerably shorter and fewer than those in model group (P < 0.05), suggesting an improvement of learning-memory ability after EA. Comparison of the ultrastructure of the neurons in the hippocampal CA 1 region showed swelling of the mitochondria, broken or disappearance of the mitochondrial cristae, degeneration of the synapses, breakage and vague outline of the basement membrane of the blood capillaries in mice of model group; and basically distinct outline of the mitochondrial cristae and microvessels, and more synaptic vesicles in EA group. CONCLUSION: EA may effectively improve the learning-memory capacity of the APP transgenic AD mice and alleviate the pathological changes of neurons of the hippocampal CA 1 region, which may be one of the mechanisms underlying the improvement of AD by EA.