| The incidence rate of metabolic syndrome (such as hyperlipidemia, diabetes,cardiovascular disease) is more and more high, and harmful to human health sincethere are the great changes of economic development, lifestyle and dietary structure,Mitochondria are the main places of cellular energy metabolism and free radicalproduction. If the metabolic process or free radical balance is destroyed, it will harmthe body and cause diseases. Many researches show that mitochondrial dysfunction isthe pathological basis of chronic metabolic diseases, such as diabetes, aging,cardiovascular disease, and cancer. Therefore, improving the quality of mitochondria,alleviating mitochondrial damage, thereby inhibiting the production of free radicals isone of the important ways to relieve chronic metabolic disease. This paper is toinvestigate the protective effect of chlorogenic acid on mitochondrial oxidativedamage and its possible mechanism in vitro and in vivo.The first part, to investigate the protective effect of chlorogenic acid onintestinal mitochondrial oxidative damage in rats in vitro.Methods: Different concentrations of H2O2damage to mitochondria,mitochondrial swelling, membrane potential and ROS were determined to ensureoptimum concentration; different concentrations of CHA into the mitochondrialsuspension, determining the CHA optimum concentration determination ofmitochondrial swelling, membrane potential and ROS. The test was divided into fourgroups: normal group, H2O2group,160μmol/LCHA+H2O2group,160μmol/LCHAgroup. The mitochondrial suspension and supernatant of cytochrome C, content of8-OHdG, T-SOD, GSH-Px, T-AOC and complexes I, IV and V activity weredetermined.Results:(1) The optimum concentration of H2O2was0.5mol/L;(2) Theoptimum concentration of CHA was160μmol/L;(3) The mitochondrial suspensioncytochrome C content of160μmol/LCHA+H2O2group was significantly increasedthan H2O2group (P<0.05);(4) The amount of8-OHdG of160μmol/LCHA+H2O2 group was significantly decreased than H2O2group (P<0.05);(5) Compared withH2O2group,160μmol/LCHA+H2O2group T-SOD, GSH-Px activity was significantlyenhanced (P<0.05), T-AOC has significant (P<0.01);(6)160μmol/LCHA+H2O2group compared with H2O2group, mitochondrial complex I, IV and V activityincreased with significant difference (P<0.01).The second part, to investigate the protective effect of chlorogenic acid onintestinal mitochondrial oxidative damage in rats in vivo.Methods:32healthy rats (180±20) g, randomly divided into four groups:normal group, TNBS group, CHA group, CHA+TNBS group,8rats in each group,the experimental period around, TNBS dose is100mg/Kg, chlorogenic acid daily bygavage, the dose of60mg/Kg. This part of the intestinal HE staining, histologicalscore, colon and ileum T-SOD, T-AOC and MDA content were determined, themitochondrial structure were observed by electron microscopy and the mitochondrialsuspension and supernatant of cytochrome C,8-OHdG content, T-SOD activity,MDA content, complex I, IV and V activity were determined.Results:(1) HE display in CHA+TNBS group, intestinal villus structure morecomplete than the TNBS group, histological grading map shows the two groups hadsignificant difference (P<0.05);(2) The activity of T-AOC in the CHA+TNBS groupstronger than in TNBS group, and T-SOD activity significant (P<0.05), the content ofMDA in CHA+TNBS group higher than TNBS group, and significant (P<0.05);(3)By the electron micrographs can be seen in TNBS group of mitochondrial membranerupture, spinal fracture or disappeared, and the fuzzy not clear reduce, mitochondrialmatrix volume, mitochondria became round swelling, those were improved markedlyin the CHA+TNBS group;(4) The mitochondrial cytochrome C content in suspension,CHA+TNBS group more increase than TNBS group;(5) In CHA+TNBS groupcomplexes I, IV and V activity is higher than that of TNBS group, and the complex ofI and V had significant difference (P<0.05).In summary, CHA has a protective effect on mitochondria from rats withintestinal injury. CHA has a protective effect on the mitochondrial complex of I, IV,and V in vitro. CHA also has a protective effect on intestinal and mitochondrial injuryin rats by increasing the antioxidant activity and protecting mitochondria injury. |
