ABSTRACT
Aim To study the protective effect of simvastatin(Sim)on liver function injury in apolipoprotein E gene knockout(ApoE KO)mice fed with high-fat diet and the underlying mechanism.Methods Twenty-four 8-week-old male ApoE KO mice were randomly divided into ApoE KO group,ApoE KO+Sim group and ApoE KO+PD150606 group.The contents of total cholesterol(TC)and triglyceride(TG)in serum and liver,and the activities of aspartate aminotransferase(AST)and alanine aminotransferase(ALT)in serum were measured.The contents of malondialdehyde(MDA)and reactive oxygen species(ROS)and the activity of superoxide dismutase(SOD)in liver were determined.The contents of tumor necrosis factor-α(TNF-α)and interleukin-6(IL-6)and the activity of calpain in liver were examined.Results Compared with C57 group,ApoE KO group showed significant increase in the contents of TC and TG in both serum and liver.In addition,the activities of AST and ALT in serum and the contents of MDA and ROS in liver significantly increased,while SOD activity in liver decreased in ApoE KO group.The contents of TNF-α and IL-6 and the activity of calpain in liver significantly increased.Compared with ApoE KO group,Sim group had no significant effects on TC and TG,while reduced the activities of AST and ALT,decreased the contents of MDA and ROS,increased the activity of SOD and decreased the contents of TNF-α and IL-6 as well as the activity of calpain in liver.PD,the calpain inhibitor,had the similar effects with Sim regarding the above mentioned parameters.Conclusions Sim improved the liver function injury of ApoE KO mice,which might be related to the inhibition of calpain activity,subsequently increasing the antioxidant capacity and reducing the inflammatory response.
ABSTRACT
OBJECTIVE: To develop the quality specification of seeds of Peganum harmala. METHODS: According to the Chinese Pharmacopoeia (2010 Version, Volume 1) and its appendix method, the water, total ash, acid insoluble ash, 50% ethanol extractives, and heavy metal were analyzed for seeds of P. harmala. TLC method was used to separate harmaline (HAL), harmine (HAR) and vasicine (VAS) in seed samples using mixture of ethyl acetate-methanol-ammonia water (10:1.5:0.5) as a developing solvent on high performance silica G pre-coated plate with 254 nm fluorescent (GF254) and to identify them inspected under UV 366 nm, 254 nm, visualized by spraying with both Dragendorff reagent and by bioautographic assay. In the HPLC method, HAL and HAR were separated on a C18 column with acetonitrile-ammonium acetate water (19:81) as the mobile phase and detected at 330 nm. The HPLC fingerprints were performed on the same C18 column and eluted by using a linear gradient of acetonitrile (A) and 0.1 mmol · L-1 ammonium acetate buffer under the flow rate at 0.7 mL · min-1 and detected at 280 nm. RESULTS: In the TLC procedures, 254 and 366 nm fluorescent, Dragendorff reagent, and bioautographic assay for the detection of acetylcholinesterase inhibitor can be used for qualitative identification of the active ingredients. For the HPLC quantitative method, the calibration curve of HAR displayed ideal linearity over the range of 1.97-198.68 μg · mL-1 with average recovery of 99.69% (RSD of 1.89%). HAL displayed ideal linearity over the range of 1.70-345.30 μg · mL-1 with average recovery of 100.66% (RSD of 1.78%). The contents of HAL and HAR in 11 batches of seeds of P. harmala were 3.234% and 3.755%. In the characteristic fingerprints of seeds of P. harmala, four common peaks were identified. CONCLUSION: The results indicated that the water, total ash, acid insoluble ash, and 50% ethanol extractives were not more than 9.0%, 8.0%, 1.0%, and 22.0%, respectively. The heavy metal of plumbum, cadmium, arsenic, mercury, and copper were not more than 5 × 10-6, 3 × 10-6, 2 × 10-6, 2 × 10-6 and 20 × 10-6, respectively. The content limit of the sum of HAL and HAR was not lower than 5.5%. With the peak of HAL as reference peak, the variance of relative retention time of the four common peaks, in the characteristic fingerprints of seeds off. harmala, should be fluctuated in the range of 5% of the specified value. The qualitative and quantitative method established was suitable for the quality evaluation and assessment of seeds of P. harmala.