| Objective: Large number of clinical data have indicated that phlegm andstasis is an important pathogenesis of non-alcoholic-fatty liverdisease(NAFLD). Danshen can improve the blood circulation and eliminatestasis.Zexie can remove water wet and reduce phlegm.They were commonlyused for NAFLD. To explore the action mechanism of DanshenZexie onNAFLD, we made reference to some relevant literatures and established theNAFLD model of rats by high fat diet feeding. At the same time, the rats wereinterfered with medicine. We observed the effect on TC, TG, FFA, SOD, MDA,ET-1,6-Keto-PGF1α, TXB2, t-PA, PAI-1and the expression ofPPARαmRNA in liver. Furthermore, we also observed the effect onpathomorphology of liver tissue by light microscope.Methods: Select60male Wistar rats, weight from180g to200g. Ratstook food and drunk water freely, and were raised in sanitary experiment labthat the temperature was18℃～22℃, bright and dim alternate every12hours.After a week, they were randomly divided into6groups: Normal group(N),Model group(M), Danshen group (A), Zexie group (B), DanZe group(C),Dong-Bao-Gan-Tai group(Y). Rats in normal group were fed with ordinaryforage and the others were fed with high-fat forage, which lasted for8weeks.Meanwhile treatment drug or control drug was given to the correspondinggroup and equal amount of saline was given to normal and model group. Thestomach-perfusion was taken once every morning and the drug volume wascalculated by1ml/100g. All rats were fasted for12hours after the laststomach-perfusion and then they were anaesthetized and their blood weredrawn out, the blood sample was made centrifugation with low temperature,and then the blood serum was sealed and stored in-20℃for standby. At thesame time, a little hepatic tissue was removed at the0.5cm borderline of the max leaf of the liver, and then it was frozen by liquid nitrogen in order todetecting the expression of PPARαmRAN. In addition, a defined part ofhepatic tissue was removed in the same position, soaked in4%paraform,paraffin fixed, sliced, HE stained to provide for the observation of the lightmicroscope. Furthermore, we observe the expression changes of t-PA andPAI-1in liver tissue with the immune chemical method.Result:1Changes of content of TC,TG,FFA in serum of NAFLD ratsCompared with normal group, the content of TC,TG,FFA in serum ofmodel group were obviously increased (P＜0.01), which showed that NAFLDrats had significant blood-fat metabolic disorder. After the intervention ofmedicine, each treatment group could significantly reduce the content of TC,TG,FFA (P＜0.01). Among them, the content of TC in Danshen group, Zexiegroup and DanZe group had no statistical significance withDong-Bao-Gan-Tai group (P＞0.05). The content of TC in DanZe group werelower than Danshen group and Zexie group (P＜0.01). Danshen group had nostatistical significance with Zexie group (P＞0.05). The content of TG inZexie group and DanZe group were lower than Dong-Bao-Gan-Tai group (P＜0.01). Danshen group had no statistical significance with Dong-Bao-Gan-Taigroup (P＞0.05). The content of TG in Zexie group were lower than Danshengroup(P＜0.01). The content of FFA in Danshen group, Zexie group andDanZe group were lower than Dong-Bao-Gan-Tai group (P＜0.01). Thecontent of FFA in DanZe group were lower than Danshen group and Zexiegroup(P＜0.01). The content of FFA in had no statistical significance withZexie group (P＞0.05).2Changes of the activity of ALT, AST in serum of NAFLD ratsCompared with normal group, the activity of AST and ALT in serum ofmodel group were markedly increased (P＜0.01). After the intervention ofmedicine, the activity of AST and ALT in serum of each treatment group wereall obviously reduced(p＜0.01). Among them, the activity of ALT in Danshengroup, Zexie group and DanZe group were lower than Dong-Bao-Gan-Tai group (P＜0.01or P＜0.05). The activity of ALT in DanZe group were lowerthan Danshen group and Zexie group (P＜0.01). The activity of AST in DanZegroup were lower than Dong-Bao-Gan-Tai group and Danshen group(P＜0.01or P＜0.05). The activity of AST in Zexie group had no statisticalsignificance with DanZe group(P＞0.05).The activity of ALT and AST inDanshen group had no statistical significance with Zexie group (P＞0.05).3Changes of the activity of SOD and the content of MDA in serum ofNAFLD ratsCompared with normal group, the activity of SOD in model group weremarkedly reduced and the content of MDA in model group were markedlyincreased (P＜0.01). Each treatment group could increase the activity of SODand reduce the content of MDA (P＜0.01). Among them, the activity of SODin Danshen group were lower and the content of MDA were higher thanDong-Bao-Gan-Tai group(P＜0.01). The activity of SOD and the content ofMDA in Zexie group, DanZe group had no statistical significance withDong-Bao-Gan-Tai group (P＞0.05). The activity of SOD in DanZe groupwere higher than Danshen group and Zexie group and the content of MDAwere lower(P＜0.01). The activity of SOD in Danshen group were lower thanZexie group (P＜0.01) and the content of MDA in Danshen group had nostatistical significance with Zexie group (P＞0.05).4Changes of content of ET,6-Keto-PGF1α and TXB2in serum ofNAFLD ratsThe contents of ET in plasma of model group were significantly higherthan normal group (P＜0.01). After the intervention of medicine, the contentsof ET in plasma of each treatment group were significantly reduced(P＜0.01).Among them, the contents of ET in Danshen group, Zexie group and DanZegroup were lower than Dong-Bao-Gan-Tai group(P＜0.01). The contents ofET in DanZe group were lower than Danshen group and Zexie group.(P＜0.01).There was no statistical significance on the content of ET betweenDanshen group and Zexie group (P＞0.05).The contents of6-keto-PGF1α in plasma of model groupwere significantly lower than normal group (P＜0.01),while the contents of TXB2inplasma of model group were significantly higher than normal group (P＜0.01).After the intervention of medicine,the contents of6-keto-PGF1α inplasma of each treatment group were significantly increased (P＜0.05or P＜0.01),while the contents of TXB2in plasma of each treatment group weresignificantly reduced(P＜0.01). There was no statistical significance amongDanshen group, Zexie group,DanZe group and Dong-Bao-Gan-Tai group(P＞0.05). The contents of TXB2in DanZe group were lower than Danshen group,Zexie group and Dong-Bao-Gan-Tai group (P＜0.05or P＜0.01). There wasno statistical significance on the contents of TXB2between Danshen groupand Zexie group (P＞0.05).5Changes of liver tissue morphology of NAFLD ratsThe live tissue of rats were observed in light microscope: In normalgroup, the structure of hepatic tissue was clear,hepatic lobules were normal,central veins were big and had thin paries. Hepatic cells were polygon whichwere arranged like cords and distributed as radial around central veins. As forthe rats in model group, the hepatic cells appeared edema and moderate tosevere fatty degeneration. In addition, a small amount of hepatic cells alsoappeared necrosis, but no significant fibrosis. Compared with the model group,the fatty degeneration in each treatment group were improved in varyingdegrees, the lipid drops of Dan-Ze group were disappeared and a few lipiddrops empty bubble were visible in Danshen group, Zexie group andDong-bao-gan-tai group.6Changes of expression of PAI-1and t-PA in liver of NAFLD ratsThe expression of t-PA were decreased and the expression of PAI-1wereincreased in model group compared with the normal group (P＜0.01). Afterthe intervention of medicine, the expression of t-PA were increased and theexpression of PAI-1were decreased in each treatment group (P＜0.01).Among them, the expression of t-PA in DanZe group were higher thanDong-bao-gan-tai group(P＜0.01).The expression of t-PA in Danshen groupno statistical significance with Zexie group (P＞0.05). The expression of PAI-1in DanZe group were lower than Dong-bao-gan-tai group(P＜0.01). Theexpression of PAI-1in Zexie group were higher than Dong-bao-gan-taigroup(P＜0.01). The expression of PAI-1in Danshen group and Zexie groupwere higher than DanZe group(P＜0.01). The expression of PAI-1in Zexiegroup were higher than Danshen group(P＜0.05).7Changes of expression of PPARαmRNA in liver of NAFLD ratsThe expression of PPARαmRNA of model rats was obviously lower thannormal group (P＜0.01). After the intervention of medicine, the expression ofPPARαmRNA of treatment group was significantly higher than model group(P＜0.01or P＜0.05).The expression of PPARαmRNA in DanZe group werehigher than Dong-bao-gan-tai group,Danshengroup and Zexie group (P＜0.01).The expression of PPARαmRNA in Zexie group were lower thanDong-bao-gan-tai group (P＜0.01) while there was no statistical significancebetween Danshen group and Dong-bao-gan-tai group(P＞0.05). Theexpression of PPARαmRNA in Danshen group were higher than Zexiegroup(P＜0.01).Conclusions：1Danshen,Zexie and the cooperation could significantly reduce thecontent of TC,TG and FFA in serum,restrain the activity of ALT,AST inserum. They had good function in regulating blood fat and protecting liver.2Danshen,Zexie and the cooperation could significantly increase theactivity of SOD and reduce the content of MDA, showing that they could clearfree radicals, enhance the ability of antioxidation, and regulate the metabolicbalance of free radicals.3Danshen,Zexie and the cooperation could significantly reduce thecontent of ET and TXB2and increase the content of6-Keto-PGF1α. They canprotect the endothelial function and improve the circulation of the liver.4Danshen,Zexie and the cooperation could obviously improve the lesiondegree of liver tissue of NAFLD rats, it manifested like this: after the drug wasgiven, the structure of hepatic lobules and sinus became clear, the fattydegeneration became significant improvement. 5Danshen,Zexie and the cooperation could restore the balance of thefibrinolytic system state through enhancing the expression of t-PA andreducing the expression of PAI-1.They can reduce blood viscosity and preventthe thrombosis.6Danshen,Zexie and the cooperation could enhance the expression ofPPARαmRNA, promote conveying of lipid, enhance oxygenolysis of fat andreduce the liver fat deposition in hepatic tissue of NAFLD to achieve thepurpose of curing the NAFLD. |
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