ABSTRACT
OBJECTIVE To investigate the improvement effect and mechanism of different extracts from Tylophora yunnanensis on non-alcoholic steatohepatitis （NASH）. METHODS Normal human liver LO2 cells were induced to steatosis by free fatty acid， then were divided into normal group， model group， silybin group （100 μmol/mL）， T. yunnanensis ethanol extracts （TYS） group （50 μg/mL）， T. yunnanensis ethyl acetate extracts （TYSA） group （50 μg/mL）， and T. yunnanensis n-butanol extracts （TYSB） group （50 μg/mL）. After 24 hours of drug intervention， the deposition of lipid droplets was observed in LO2 cells in each group. The contents of total cholesterol （TC）， triacylglycerol （TG）， malondialdehyde （MDA） and glutathione （GSH）， the activities of aspartate transaminase （AST）， alanine transaminase （ALT） and superoxide dismutase （SOD）， the mRNA expressions of Kelch-like ECH-associated protein 1（ Keap1）， nuclear factor E2-related factor 2（ Nrf2） and heme oxygenase 1（ HO- 1） were detected. NASH rat model was induced by a high-fat diet， and then divided into normal group， model group， silybin group （12.6 mg/kg）， TYS group （80 mg/kg）， TYSA group （80 mg/kg） and TYSB group （80 mg/kg）， with six rats in each group. The liver indexes of rats in each group were calculated after 6 weeks of drug intervention. The liver histopathological changes were observed， and the contents of TC， TG， HDL-C and LDL-C， AST and ALT activities in serum， the contents of MDA and GSH， SOD activities in liver tissue were detected. RESULTS Compared with model group， TYS， TYSA and TYSB could reduce lipid droplet deposition， intracellular TC， TG and MDA contents， AST and ALT activities， and increase SOD activity， GSH content， and Keap1， Nrf2， HO-1 mRNA expression levels in LO2 cells after steatosis to varying degrees， with some differences being statistically significant （P＜0.05）. They also significantly improved liver injury in NASH model rats， reduced their liver indexes， TC， TG， LDL-C and MDA contents， AST and ALT 1-042） activities， and increased HDL-C （except for TYS and TYSB）， GSH contents and SOD activity， with TYSA having the most significant effect （P＜0.05）. CONCLUSIONS TYS， TYSA and TYSB have a certain improvement effect on NASH， among which TYSA has the most obvious effect. Its mechanism of action may be related to upregulating the Keap1/Nrf2/HO-1 signaling pathway and inhibiting oxidative stress
ABSTRACT
OBJECTIVE To study the improvement effects of different polar parts fro m total f lavonoids of Scutellaria amoena on non-alcoholic fatty liver disease （NAFLD）model rats. METHODS The total flavonoids of S. amoena （SAF）were extracted by reflux extraction with ethanol ，suspended with water ，and then extracted with ethyl acetate and n-butanol in order to obtain the extraction parts of SAF （recorded as SAFA and SAFB respectively ）. Thirty-six rats were randomly divided into normal group （n＝ 6）and modeling group （n＝30）. Modeling group was given high-lipid diet to induce NAFLD model. After modeling ，modeling group was randomly divided into model group （normal saline ），fenofibrate group （positive control ，20 mg/kg），SAF group （300 mg/kg），SAFA group （300 mg/kg）and SAFB group （300 mg/kg）；they were given relevant intragastical administration ，once a day，for consecutive 6 weeks. After last administration ，the liver index was calculated ；the levels of total cholesterol （TC）， triacylglycerol（TG），aspartate transaminase （AST），alanine transaminase （ALT），high-density lipoprotein cholesterol （HDL-C）， low-density lipoprotein cholesterol （LDL-C） in serum ，the levels of superoxide dismutase （SOD），glutathione peroxidase （GSH-Px），malondialdehyde（MDA），interleukin-1β（IL-1β），IL-6 and tumor necrosis factor-α（TNF-α）in liver tissue were detected；the pathomorphological changes of liver tissue were observed. RESULTS Compared with normal group ，the liver index ， the levels of TC ，TG，AST，ALT，LDL-C，MDA，IL-1β， IL-6 and TNF-α in serum/liver tissue of model group were all increased significantly （P＜0.05）， while the levels of HDL-C，SOD and GSH-Px were all decreased significantly （P＜0.05）. Compared with model group ，except there was no statistical significance in the serum levels of HDL-C and ALT in SAFA group （P＞0.05），above indexes in serum/liver tissue of rats in groups of polar parts from total flavonoids of S. amoena were significantly improved （P＜0.05）；inflammatory cell infiltration and fatty vacuoles in liver tissue were significantly improved. Compared with SAF group and SAFA group ，the levels of TC，TG，AST，MDA，IL-6 and TNF-α were decreased significantly in SAFB group（P＜0.05），while the level of SOD was increased significantly （P＜0.05）；pathomorphological changes of liver tissue were improved more significantly. CONCLUSIONS Each polar part from total flavonoids of S. amoena can improve NAFLD by regulating oxidative stress and inhibiting the secretion of inflammatory factors. The n-butanol polar part has more obvious effect .
ABSTRACT
OBJECTIVE：To study the antioxidan t activity and lipid-lowering effect of ethanol extract and its different solvent extracts from the stems and leaves of Scutellaria amoena . METHODS ：The stem and leaves of S. amoena was extracted with 95％ ethanol to obtain ethanol extract ，and then extracted with petroleum ，ethyl acetate and n-butanol to obtain corresponding different solvent extracts. Using vitamin C （Vc）as positive control ，the antioxidant activities of ethanol extract ，petroleum ether extract ， ethyl acetate extract and n-butanol extract from the stems and leaves of S. amoena were determined by hydroxyl radical ，superoxide anion radical and DPPH radical scavenging method ，and the IC 50 was calculated. Steatosis L 02 hepatocyte model was established with fat emulsion. Using fenofibrate （20 μg/mL）as positive control ，the effects of high and low concentration （100 and 50 μg/mL） ethanol extract ，ethyl acetate extract and n-butanol extract from the stems and leaves of S. amoena on the contents of TC and TG in cells were investigated. RESULTS ：The order of scavenging ability to hydroxyl radicals was n-butanol extract ＞ethyl acetate extract＞Vc＞ethanol extract ＞petroleum ether extract ；IC50 of them were 0.15，0.17，0.35，0.75，1.17 mg/mL，respectively. The order of scavenging ability to superoxide anion radical was Vc ＞n-butanol extract ＞ethyl acetate extract ＞ethanol extract ＞ petroleum ether extract ；IC50 of them were 0.034，0.55，0.75，3.32，3.73 mg/mL，respectively. The order of DPPH scavenging ability to DPPH radical was Vc ＞n-butanol extract ＞ethyl acetate extract ＞ethanol extract ＞petroleum ether extract ；IC50 of them were 0.003 2，0.028，0.033，0.048，0.057 mg/mL， respectively. The ethanol extract ，ethyl acetate extract and n-butanol extract from the stems and leaves of S. amoena could significantly decrease the contents of TC and TG in steatosis L 02 hepatocytes （P＜0.01）. The order of lipid-lowering ability was n-butanol extract （low dose ）≈fenofibrate＞ethyl acetate extract （high dose ）＞ethanol extract （high dose ）＞ n-butanol extract （high dose ）＞ethyl acetate extract （low dose ）＞ethanol extract （low dose ）. CONCLUSIONS ：The ethanol extract ， petroleum ether extract ，ethyl acetate extract and n-butanol extract from the stems and leaves of S. amoena show good antioxidant activity and lipid-lowering effect （except for petroleum ether extract ）. Ethyl acetate extract and n-butanol extract possess the strongest antioxidant activity and lipid-lowering effect.