Study On Pathogenesis Of Oxidative Damage And Energy Metabolism Of Mitochondria In Damp-heat Syndrome Of Seasonal Febrile Disease And Effects Of Chinese Herbs

Background:Damp-heat syndrome of seasonal febrile disease is common clinical disease, involving pathological changes of many systems and organs and tissues. Because of the special climate, geography, food habit and constitution factors in Ling Nan Area, damp-heat disease is common and easy, so it is important significance to study damp-heat syndrome in Ling Nan Area. The current researches demonstrate that pathogenesis of damp-heat syndrome of seasonal febrile disease is not limited to one entrails or target, it is related to immunologic function, inflammation, aquaporins, helicobacter pylori, gastrointestinal motility, endotoxin, oxidative damage, microecology, trace elements, blood lipid metabolism and energy metabolism, etc. On the basis of clinical and experimental research, we explore the pathogenesis of damp-heat syndrome of seasonal febrile disease from oxidative damage and energy metabolism of mitochondria and effects of Chinese herbs.Objective:The research established mouse model of damp-heat syndrome of seasonal febrile disease through richly fatty and sweet diet, humid heat environment and infected by MHV-A59, in which MHV-A59 was used as biological pathogenic factor. The research studied the pathogenesis of damp-heat syndrome of seasonal febrile disease from oxidative damage and energy metabolism of mitochondrion in the model mouse, by observing the ultramicrostructure, superoxide dismutase(SOD), glutathione proxidase (GSH-PX), malondialdehyde(MDA), membrane potential, ATPase activity, ATP concent and activities of respiratory chain complexⅠ,Ⅱ,Ⅲ,Ⅳin the hepatic mitochondrion of mice model of damp-heat syndrome of seasonal febrile disease. The research certificated the pathogenesis of damp-heat syndrome of seasonal febrile disease by treatment with Decoction of Hao qin qing dan and Decoction of Gan lu xiaodu, observed the treatment mechanism of clearing away heat and dampness. It was supplementary and evidence for pathogenesis of damp-heat syndrome of seasonal febrile disease by studing SOD, GSH-PX and MDA of peripheral blood of patients with exogenous damp-heat disease.Methods:1. Literature research:on the basis of the approach of evaluating and reviewing articles, we proposed to arrange the study state of animal model of damp-heat syndrome of seasonal febrile disease, study the survey of essence of damp-heat syndrome, the modern study of oxidative damage and energy metabolism of mitochondrion, sum up the study progression of MHV and the relationship between therapy of clearing away heat and dampness and oxidative damage and energy metabolism of mitochondrion.2. Experimental research:①establishment of damp-heat syndrome of seasonal febrile disease model. four-weeks-old male mice of BALB/c was divided randomly into four groups:control group(normal diet), simple damp-heat syndrome group(richly fatty, sweet diet and humid heat environment), simple virus infection group (normal diet and virus infection) and model group (richly fatty, sweet diet, humid heat environment and infected by MHV-A59), The triglyceride (TG), the indexes of liver function ALT,AST and Hepatic histopathology in mice were detected by biochemistry and ELISA. In the experiment we observed the symptom and sign of mice.②pathogenesis of damp-heat syndrome of seasonal febrile disease:four-weeks-old mice of BALB/c was divided randomly into four groups:control group, simple damp-heat syndrome group, simple virus infection group and model group. The ultramicrostructure of hepatic mitochondria in each group was observed with electronic microscope and the change of SOD, GSH-PX, MDA, membrane potential, ATPase activity, ATP content and activities of respiratory chain complexⅠ,Ⅱ,Ⅲ,Ⅳin the hepatic mitochondrion of mice were detected.③The pathogenesis of treating damp-heat syndrome of seasonal febrile disease with the decoction of clearing away heat and dampness:four-weeks-old mice of BALB/c were divided into four groups randomly, model group, Hao qin qing dan decoction group (mice of model group were treated by Decoction of Hao qin qing dan), Gan lu xiao du decoction group(mice of model group were treated by Decoction of Gan lu xiao du), western medicine group (mice of model group were treated by Polyene Phosphatidylcholine Capsules).The change of SOD, GSH-PX,MDA, membrane potential, ATPase activity, ATP content and activities of respiratory chain complexⅠ,Ⅱ,Ⅲ,Ⅳin the hepatic mitochondrion of mice were detected3. Clinical research:The 30 patients of exogenous damp-heat syndrome as the test group were from out-patient clinic of guangzhou hospital of Chinese medicine. There were 25 healthy people undergoing medical examination as control group. The level of SOD, GSH-PX and MDA of patients and healthy people were detected by colorimetry. The patients were treated by Decoction of Hao qin qing dan for three days. Then we detected the level of SOD, GSH-PX and MDA again. Healthy people without treatment.Results:1.In the research of establishment of damp-heat syndrome of seasonal febrile disease model:Compared to control group, the mice in model group showed some features such as drinking less water, eat less food, viscous and more plentiful excretion, weight loss, rectal temperature going up, bulky hair, etc. These signs and symptoms were closely related to the features of damp-heat syndrome of seasonal febrile disease. The level of TG obviously risen up compared to control group and simple virus infection group (p＜0.01). The liver of the mice under microscope in model group showed vacuolating chang, inflammatory cell infiltration,100% leaflet inflammation, hepatic steatosis, necrosi. Hepatic steatosis was not existing in control group and simple virus infection group. These pathological changes were closely related to basically pathological features of damp-heat syndrome of seasonal febrile disease.2. In the research of oxidative damage of mitochondria:The level of SOD, GSH-PX and membrane potential decline and MDA risen up in model group obviously compared to control group, simple damp-heat syndrome group and simple virus infection group (P＜0.01 or P＜0.05). These show that lipid peroxidation content risen up and relative antioxidase activity decreased, mitochondrion function was injured in damp-heat syndrome of seasonal febrile disease model. Decoction of Hao qin qing dan and Decoction of Gan lu xiao du could improve the activities of SOD, GSH-PX and membrane potential in the hepatic mitochondria of damp-heat syndrome of seasonal febrile disease, improve the function of defense system scavenging free radicals, at the same time,they reduced MDA content and scavenged the free radicals. The difference was significant compared to model group (p＜0.01) These findings indicate that Chinese herbs of clearing away heat and dampness can reinforce mitochondria function of antioxidation and reduce the injure of oxidation to protect mitochondrion though improving the function of defense system scavenging free radicals and reducing the production of free radicals. There is no significant difference in improving the activities of SOD, GSH-PX and membrane potential in the hepatic mitochondrion between model group and western medicine group, but western medicine group obviously ruduce MDA content compared to model group(P＜0.05). The research result indicate that the mechanism of Chinese herbs clearing away heat and dampness was to improve the activities of SOD, GSH-PX and mitochondrial membrane potential, improve the function of defense system, reduce the production of free radicals, remove the producted free radicals in treatment of damp-heat syndrome of seasonal febrile disease.3. In the research of energy metabolism:The level of ATPase activity and ATP content obviously decline in model group and simple damp-heat syndrome group compared to control group, specially in model group (P＜0.01).The decreasement of ATPase activity indicates that energy decomposition and utilization decrease. The decreasement of ATP content indicates that energy product Insufficiently. The research result indicate that energy metabolize disorderly and the production and utilization of ATP decrease in damp-heat syndrome of seasonal febrile disease. Chinese herbs clearing away heat and dampness could improve the ATPase activity and ATP content obviously compared to model group (P＜0.01), improve energy metabolic disorders and promote the production of energy.Activities of respiratory chain complexⅠ,Ⅱ,Ⅲ,Ⅳin the hepatic mitochondrion of mice from model of damp-heat syndrome of seasonal febrile disease decline obviously compared to control group (p＜0.01), which illustrate that serious energy metabolic disorders and dysfunction of mitochondrion may esist in damp-heat syndrome of seasonal febrile disease. Activities of complexⅠ,Ⅱ,Ⅲ,Ⅳin Hao qin qing dan decoction group and Gan lu xiao du decoction group obviously risen up compared to model group (p＜0.05 or p＜0.01). The research result indicate that Chinese herbs clearing away heat and dampness could improve the activities of complexⅠ,Ⅱ,Ⅲ,Ⅳ, in order to promote the production and utilization of energy, improve the energy metabolism of mitochondrion.4. clinical research:Compared to healthy people, the level of Peripheral blood SOD, GSH-PX of patients obviously decreased (p＜0.01) and MDA rised up (p＜0.01), there were significant differences. The level of Peripheral blood SOD, GSH-PX of patients who treated by decoction of Hao qin qing dan obviously rised (p＜0.01 or p＜0.05), and MDA obviously decreased (p＜0.01).Conclusion:1.A new mouse model of damp-heat syndrome of seasonal febrile disease was duplicated by richly fatty and sweet diet, humid heat environment and MHV-A59. MHV-A59 was a biological pathogenic factor in the model which possessed characteristics of the illness and syndrome.2.Oxidative damage and energy metabolic disorders exist in damp-heat syndrome of seasonal febrile disease. Chinese herbs clearing away heat and dampness could reduce oxidative damage and improve production and utilization of energy, which was in favor of recovery of normal function of cell. So, oxidative damage and energy metabolic disorders in mitochondrion may be one of the pathogenesis of damp-heat syndrome of seasonal febrile disease.3. Both decoction of Hao qin qing dan and decoction of Gan lu xiao du could reduce oxidative damage of mitochondrion to a certain extent in the damp-heat syndrome of seasonal febrile disease, protect the function of mitochondrion, improve the energy metabolism. These proof the pathogenesis of damp-heat syndrome of seasonal febrile disease discussed above. Chinese herbs of clearing away heat and dampness could reduce oxidative damage of mitochondrion and promote the production and utilization of energy.4.Oxidative damage and antioxidant enzyme system function deficiency exists in the body with damp-heat syndrome of seasonal febrile disease. Decoction of Hao qin qing dan could reduce the oxidative damage in treating viral upper respiratory infection of damp-heat syndrome...