The Morphological Observation Of Lymphatic Stasis Effect On Hepatic Tissue Leptin Immunoreactive Cells

Objective:The liver is the organ of the body one of the most important, not only in terms of metabolism and body tissues and organs are closely related, but also with the secretion, excretion, and biotransformation, especially in fat metabolism, the liver plays an important role. Leptin is obese gene (ob gene) encoding product. In fat metabolism has an important role, such as appetite suppression, decreased energy intake, increased energy consumption, inhibit fat synthesis. As is known to all, lymphatic vessels is a kind of one-way drainage vessel systems, mainly recycles macromolecular substances such like liquid, fat and protein, etc. The liver can produce a large number of lymph, Nearly half of thoracic duct lymph from the liver. Understanding the regulation of fat metabolism in the liver and the leptin metabolism and Intrahepatic lymphatic role in the metabolism of the body, These have important physiological and pathological significance. To investigate the relationship between leptin and liver and lymph stasis on the morphology of liver tissue to provide morphological evidence, The immunoreactive cells of hepatic tissue in normal SD rat and animal models of thoracic duct ligation were observed in the study.Methods:1 Experimental Animal and MaterialsThe 30 rats were used, body weight 250 ~ 300g, present from the Experimental Animal Center of Hebei Medical University, either male or female, were randomly divided into experimental (operation= 20) and control (sham operation = 10) groups.Before 2 hours on the experiment surgery, laboratory animals feeding fatty foods, and then at 6% chloral hydrate (5ml/kg) intraperitoneal injection of anesthesia, the experimental group of thoracic duct ligation in the abdomen, while the control group not only full of free thoracic duct ligation. After operation, experimental animals fed conventional, drawn six months for light and electron microscopy preparation of tissue samples.2 Sample preparation and light microscope observationThe hepatic tissue sections embedded with paraffin, routine HE staining and light microscopic observation; to show the fine structure of hepatic tissue and intrahepatic distribution of vascular system.3 Immunohistochemical staining and light microscope observationThe hepatic tissue of Control and experimental group in the SD rat paraffin sections, it carried out immunohistochemical staining of leptin, then leptin and its receptor observed by light microscopy immunohistochemistry results.4 Preparation and observation for the transmission electron microscope samplesFor the ultra-structure characteristics of the SD rat hepatic tissue, and the influence on the hepatic tissue ultra-structure of the experimental lymph silted up, we need to prepare the hepatic tissue sample for transmission electron microscope observation, on the ultra-thin tissue section of rat liver.Results:1 Because of preoperative administration of lipid fed animals, a large number of chyle into the mesenteric lymphatic vessels material to run, making the mesenteric lymph nodes, intestinal trunk, into the thoracic duct were white, we easily observed.On the bottom of the ligation site, a lot of milky chyle material gathered here, making this part of the thoracic duct filling, expansion, wall thinning, translucent; and ligation of the lower part of the top there is no significant change.2 On HE staining section of hepatic tissue in the control group, we can be observed that the structural integrity of the liver, the central vein is located in the center of hepatic lobule, hepatic cord-like plates were brought together to the central vein, hepatic sinusoids in the hepatic cord between The gap between the hepatic plates and sinusoids connected to each other, hepatic cells and liver sinusoidal structure is more clear. On the liver portal area, showing interlobular artery, interlobular vein and interlobular bile ducts of three main channels.The six months after the operation, paraffin sections of hepatic tissue stained with HE, In the microscope we observed that Lobule's central vein irregular shape, expansion, more than a fracture, which was widened near the hepatic sinusoids, hepatic cord disarrangement Around the deformated central vein, we can see that lipid droplets of varying sizes in the liver cells and number of vacuolated fat cells.3 Experimental results show that in the control group SD rat's liver tissue, the immune response of leptin appears uniform positive results, there are brown colored granules. Near the central vein and hepatic portal area of the cells is slightly stronger tan colored. Experimental observations show that strong immune responses in liver tissue of leptin-positive brown colored granules increased significantly compared with the control group, the central vein and hepatic portal area is relatively large, and in the experimental group, fat cells and lipid droplets within the liver cells near the liver tissue of leptin also see strong positive immune response to tan coloredIn SD rats of model group, positive expression of leptin in Hepatic tissue of regional compared with the control group, it is a statistically significant4 TEM can observed, the liver cell membrane structure clear, rich in cytoplasm and less dense material, the nucleus is round or oval, thin nuclear membrane, and the continuous and complete. Near the Disse space of liver sinusoids may have a small amount of fibrous connective tissue, but also shows a small number of microvilli, but no basement membrane.Communication between the two samples of liver cell gap, is the basic structure of tiny bile duct. In the wall of hepatic sinusoid, it show that partial flat endothelial cells, we can also be found kupffer cells, larger, irregular in shape with cytoplasmic processes, most of the cytoplasm sudden perisinusoidal space, a small part of the cytoplasm can be attached to the endothelium or through the endothelium into the hepatic sinusoid.It is significant changes in liver cells, that nuclear membrane thickening, discrete, nuclear membrane pore increases, the nuclear chromosomes gathered into one, uneven distribution of the nuclear matrix. In the liver cytoplasm, we can be found the cytoplasm becomes empty, the cell matrix increased, the electron density reduction and lipid deposition, like edema and fatty degeneration of liver cell performance. In the perisinusoidal space, we can be found in lipid deposition and increased connective tissue fibrous. Between cells in the liver we can also be found in large deposits of lipid material and a proliferation of connective tissue fibrous. Also can found that apoptosis of liver cells.Conclusion:1 In physiology, the liver can occur immunoreactive positive expression for leptin, after ligation of thoracic duct, liver expression of leptin can enhance the immune response. Therefore, leptin and liver are functionally closely linked.2 Produced primarily lymph by the liver return to thoracic duct,to legate SD rat thoracic duct in the abdomen,liver lymph stasis animal model can be successfully established.3 The thoracic duct ligation can cause the liver lymphatic flow dysfunction,Lymphatic stasis can be induced the lipid deposition in hepatic tissue and fatty changes of rat liver.