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BACKGROUND: Among multiple signals which affect the function of melanocyte, nitrogen monoxide (NO) has been thought as an important signal molecule. Emodin hypo-acid, effective component of rhubarb, can affect proliferation of melanocyte and regulate activity of tyrosinase in cells. OBJECTIVE: To study the effect of emodin hypo-acid on expression of nitricoxide synthase (NOS) of melanocyte in skin of guinea pig and definite the effective concentrations and mechanism of melanocyte in synthesizing melanin in living skin.DESIGN: Completely randomized grouping design and controlled study.SETTING: Department of Dermatology, Affiliated Hospital of Luzhou Medical College.MATERIALS: The experiment was completed at the Infection Laboratory of Affiliated Hospital of Luzhou Medical College and Laboratory of Histology and Embryology of Luzhou Medical College from January to June 2004. A total of 24 adult healthy male guinea pigs were randomly divided into 6 groups: control group, 2, 5, 10, 20 and 40 mg/L emodin hypo-acid groups with 4 in each group.METHODS: ① Emodin hypo-acid was diluted with dimethyl-sulfoxide into 2, 5, 10, 20 and 40 mg/L, then guinea pigs in emodin hypo-acid groups were injected subcutaneously with relevant dosages of emodin hypoacid which was provided by National Institute for the Control of Pharmaceutical and Biological Products into local skins of haunch and back, and the injected volume was 1 mL. Twenty-four hours later, 1 mL emodin hypo-acid of the same concentration was injected once again into one of the places which was injected before. Skin samples were selected after 48 hours and routine paraffin section was made. Skins which were not injected with any drugs were selected from control group and emodin hypoacid groups to regard as experimental control group. ② Expression of NOS was assayed with immunohistochemical method and melanocyte was identified under high-times optic microscope to observe stain and characteristic of cytoplasm. Five sections were randomly selected from each group. Every 20 cells on each section were measured with MLAS-1000 imaging analyzer and computer processing system was used to calculate average absorbency (A) of positive immune product in melanocyte. ③ Measurement data were compared with analysis of variance, and differences among groups were compared with SNK-q.MAIN OUTCOME MEASURES: Expression of NOS in melanocyte was assayed with immunohistochemical method and results were obtained with optic microscope and imaging analyzer.RESULTS: Average A value of positive immune product in melanocyte was lower in 2, 5, 10, 20 and 40 mg/L emodin hypo-acid groups than that in experimental control group (0.126±0.118, 0.103±0.082, 0.118±0.097,0.122±0.095, 0.112±0.078, 0.196±0.066, P ＜ 0.05). There was no significant difference among emodin hypo-acid groups at various concentrations.CONCLUSION: ① Emodin hypo-acid can regulate expression of NOS in melanocyte through NO signal transduction pathway. ② Expression of NOS is not changed as the mass concentration of emodin hypo-acid varied from 2 to 40 mg/L.
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BACKGROUND: The c-fos gene is commonly expressed in neurons,which may act as one of the signs of activities and reflection of injured neural cells.OBJECTIVE: To investigate the expression of hippocampal c-fos gene of rats with ischemic cerebral injury and the protective role of shenmai injection at the molecular level.DESIGN: Randomized controlled study.SETTING: Department of Histology and Embryology, Luzhou Medical College.MATERIALS: The experiment was completed at the Department of Histology and Embryology, Luzhou Medical College from January to March 2002. Totally 30 male Wistar rats were randomly divided into control group, ischemia group and treatment group with 10 in each group.METHODS: Bilateral common carotid arteries of rats in the ischemia and treatment groups were splinted for 30 minutes and reperfused for 1 hour to establish models of ischemic cerebral injury. Rats in the treatment group were injected with 2 mL/kg of shenmai (constituted with hongshen, dwarf lilyturf tuber and other Chinese herbal medicines) 30 minutes before ischemia. Rats in the ischemia group were not treated with any drugs, and rats in the control group were treated with sham operation but without splinting common carotid artery and giving any drugs. Hippocampal tissue of rats was obtained to make paraffin sections. In every group, one section from each rat was taken at random. Totally 100 neurons of every group were counted. Expression of c-fos gene in hippocampal neuron was observed according to the nuclear color of neurons (+++ as dark-brown mark;++ as brown mark; + as light brown mark; - as no brown mark).MAIN OUTCOME MEASURE: c-fos expression of hippocampal neurons of rats in each group.RESULTS: Totally 30 rats entered the final analysis. Expression of c-fos gene in hippocampal neurons was significantly more in the ischemia group than that in the control group (+++: 24/visions, 7/visions, P ＜ 0.05); but that in the treatment group was less than that in the ischemia group (+++:13/visions, 24/visions, P ＜ 0.05).CONCLUSION: Shenmai injection can reduce the expression of c-fos gene in hippocampal neurons of rats with ischemic cerebral injury, and can protect neural cells of ischemic cerebral injured tissue.
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Objective To observe the change of the precentral gyrus blood-brain barrier ultrastructure in the epilepsy rats induced by coriaria lacton and affected by topimmate,and to study the effect of the blood-brain barrier on the mechanism of the falling sickness and therapy by topimmate.Method Thirty healthy male Sprague Dawley rats were randomly divided to three groups: the control group,the epilepsy group and the treatment group.The coriaria lacton was injected into the lateral ventricles of the rats in the epilepsy and treatment groups,and the epilepsy animal model was established.Gastric perfusion was performed 1 hour after falling epilepsy to the rats of the treatment group and was repeated one time per day for 7 days.The precentral gyrus was cut 7days later,and the ultrathin sections were made.The change of blood-brain barrier ultrastructure was observed under electron microscope.Results The endothelium,basement membrane,pericyte of epilepsy group displayed obvious edema.The electron density reduced,and the edema of the treatment group alleviated.There was signifcant difference between the epilepsy group and the control group(P