Distribution of nitric oxide synthase positive neurons in the substantia nigra of rats with liver cirrhosis

Abstract

Background Nitrogen monoxide plays an important role in the physiological activity and pathological process of striatum in substantia nigra, and the nitric oxide synthase in substantia nigra may have characteristic changes after liver cirrhosis. Objective To observe the distribution and forms of nitric oxide synthase (NOS) positive neurons and fibers in substantia nigra of rats with liver cirrhosis. Design A comparative observational experiment. Settings Beijing Friendship Hospital; Capital Medical University. Materials Twenty 4-month-old male Wistar rats (120–150 g) of clean grade, were maintained in a 12-hour light/dark cycle at a constant temperature with free access to standard diet and water. Cryostat microtome (LEICA, Germany); All the reagents were purchased from Sigma Company. Methods The experiment was carried out in the Department of Anatomy (key laboratory of Beijing city), Capital Medical University from July 2000 to March 2002. The rats were randomly divided into normal group ( n =10) and liver fibrosis group ( n =10). Rats in the liver fibrosis group were subcutaneously injected with 60% CCl 4 oil at a dose of 5 mL/kg for the first time, and 3 mL/kg for the next 14 times, twice a week, totally 15 times. Liver fibrosis of grades 5–6 was taken as successful models. Whereas rats in the normal group were not given any treatment. Four months after CCl 4 treatment, all the rats were anesthetized to remove brain, and frontal frozen serial sections were prepared. The expressions of nitric oxide synthase positive neurons in substantia nigra of rats were observed under inverted microscope. The number and gray scale of cell body of nitric oxide synthase positive neurons in substantia nigra were detected with NADPH-diaphorase staining. Main outcome measures ▪ Number and gray scale of cell body of nitric oxide synthase positive neurons in substantia nigra; ▪ Expressions of nitric oxide synthase positive neurons in substantia nigra. Results All the 20 rats were involved in the analysis of results. ▪ The nitric oxide synthase positive neurons in substantia nigra were obviously fewer in the liver cirrhosis group than in the normal group ( P < 0.01), and the gray scale of the positive cell body was higher in the liver cirrhosis group than in the normal group ( P < 0.05). ▪ Abundant nitric oxide synthase positive neurons were observed in substantia nigra of normal rats, the cell body of nitric oxide synthase positive neurons was clear and transparent, with short own cloudy processes. In substantia nigra of rats with liver cirrhosis, the body of nitric oxide synthase positive neurons were observed shrink obviously, less fibrin than normal. Conclusion Rats with liver cirrhosis may suffer from the physiological dysfunction of neurons due to lack of fibers. The nitric oxide synthase positive neurons in substantia nigra can shrink and reduce.