Effects And Mechanisms Of Pure Cannabis Sativa L.juice In Hyperlipidemia Rats

Objective:In this study,SD rats were fed with high-fat diet to establish an animal model of hyperlipidemia.The effects of pure Cannabis sativa L.juice(CSJ)on body weight,liver index,serum biochemical indexes and liver pathological morphology were observed to clarify its lipid-lowering effect;The effects of CSJ on serum inflammatory factors,oxidative stress factors,liver sterol regulatory element binding protein-1(SREBP-1),peroxisome proliferator activated-receptor γ(PPARγ)and 3-hydroxy-3-methyl glutaryl coenzyme A reductase(HMGCR)were observed protein expression.in liver and objective to explore the lipid-lowering mechanism of hemp seed drink.Methods:1.Component analysis of CSJCSJ samples were prepared by ultrasonic extraction with methanol.The content of tetrahydrocannabinol,cannabinoid and amino acid in CSJ was determined by HPLC.2.Experimental study on lipid lowering effect of CSJMale SD rats were fed with self-made high-fat diet for 16 weeks to establish hyperlipidemia model.60 animals were randomly divided into 6 groups,10 in each group.They were blank control group,model group,positive drug control group(simvastatin),CSJ low-dose,medium dose and high-dose groups.At the same time,the rats were given intragastric administration,and their body weight was measured once a week.The right lobe of liver was removed and fixed in 4% paraformaldehyde.HE staining was used to observe the pathological changes of liver;The left medial lobe of liver was stained with oil red to observe the lipid accumulation.At the same time,blood samples were taken from abdominal aorta to separate serum.The contents of TC,TG,LDL-C,HDL-C,FFA,ALT and AST were determined according to the instructions of the kit.3.The mechanism of CSJ on Hyperlipidemia RatsThe levels of IL-6,TNF-α,MDA,GSH and SOD in serum of hyperlipidemia model rats were determined by ELISA or microplate method.The protein SREBP-1,HMGCR and PPARγ were detected by Western blot.Result:1.Component analysis of CSJThe content of cannabinol in CSJ was 0.51% μ It contains crude protein(0.75-0.99%)and 16 kinds of amino acids.2.Study on the lipid-lowering effect of CSJThe body weight data of rats in each group increased steadily,and there was no difference between groups(P > 0.05).Compared with the model group,the liver index of CSJ medium dose group was significantly decreased(P < 0.05),Compared with the model group,the liver index of CSJ medium dose group was significantly decreased(P <0.05),and the liver index of CSJ high and low dose groups was decreased(P > 0.05)Compared with the blank group,the contents of TC,TG and LDL-C in the model group were significantly increased(P < 0.05,P < 0.01),the contents of HDL-C were decreased(P < 0.01),and the contents of FFA were also increased(P > 0.05),indicating the success of the model;Compared with the model group,the serum TC content of CSJ low,medium and high dose groups decreased(P < 0.05),the HDL-C content increased(P< 0.05),and the TG content of CSJ medium dose group decreased(P < 0.05).LDL-C and FFA contents were decreased in low,medium and high dose of CSJ(P > 0.05).HE staining: compared with the blank group,the liver cells in the model group were turbid,swollen and degenerative,with edema,steatosis,balloon like changes and obvious inflammatory infiltration.Compared with the model group,after CSJ treatment,the pathological changes of liver tissue were alleviated,the structure of hepatic cord was clear,the degree of hepatocyte steatosis was significantly improved,the number of vacuoles and inflammatory reaction were alleviated.Oil red staining: compared with the blank group,the lipid droplets in the liver tissue of the model group were densely distributed in the liver tissue,indicating that there was obvious lipid accumulation in the liver of the model group.Compared with the model group,the distribution and quantity of lipid droplets in liver tissue were decreased after CSJ treatment,which indicated that CSJ could improve the lipid accumulation in liver tissue.Compared with the blank group,the levels of alt,AST and AST / ALT in the model group were significantly increased(P < 0.01).Compared with the model group,the contents of ALT and AST in low,medium and high dose CSJ groups were significantly decreased(P < 0.01),and the ratio of AST / ALT was significantly decreased(P < 0.01,P< 0.05,P < 0.01),suggesting that CSJ can improve the liver function of hyperlipidemia rats.3.The mechanism of CSJ on Hyperlipidemia RatsCompared with the blank control group,the serum IL-6 and TNF levels in the model group were significantly higher-α Compared with the model group,the levels of IL-6 and TNF in CSJ low,medium and high dose groups were significantly higher(P < 0.05,P <0.01)-α The content of CSJ decreased significantly(P < 0.05,P < 0.01),suggesting that CSJ can inhibit the inflammatory reaction induced by hyperlipidemia.Compared with the blank group,the content of MDA in the model group increased(P< 0.01),the content of GSH decreased(P < 0.01),and the content of SOD also decreased(P > 0.05).Compared with the model group,the MDA content in the low,medium and high dose groups of CSJ was significantly decreased(P < 0.01),and the GSH level was significantly increased(P < 0.05).In addition,the SOD content in the low,medium and high dose groups of CSJ had an upward trend(P > 0.05),suggesting that CSJ has an obvious antioxidant effect.Compared with the blank group,the protein expressions of SREBP-1 and HMGCR in the model group were significantly increased(P < 0.05).And the expression of PPARγ was increased(P > 0.05).Compared with the model group,the expression of SREBP-1 in the liver of CSJ medium and high dose groups was decreased(P < 0.05),the expression of HMGCR protein in CSJ medium and low dose groups was decreased(P < 0.05),and the expression of PPARγ in the liver of rats was decreased,there was no significant difference in protein expression among CSJ groups(P > 0.05).Conclusion:1.CSJ can reduce the levels of TC and TG in serum,increase the content of HDL-C,improve the pathological morphology of liver,and reduce the content of ALT and AST in serum of hyperlipidemic rats,2.The lipid-lowering mechanism of CSJ may be related to the inhibition of inflammation and oxidative stress in vivo,down-regulation of SREBP-1 and HMGCR protein in liver tissue,and then inhibition of lipid production and accumulation in vivo.