| Bao-Yuan-Ling(BYL),an apitherapy formulation followed by the traditional Chinese medicine formulation principle "monarch,minister,assistant and guide",is a compound preparation of bee products including propolis,royal jelly and bee venom.Besides the anti-tumor effects of BYL in vivo and in vitro,improvement effects of myocardium in heart failure animal models and frog hearts in vitro have been demonstrated.Although BYL has been applied in clinic with the anti-hypertension effect,the mechanism was still ambiguous.The purpose of this paper was to explore the effect and mechanism of BYL on blood pressure and cardiac hypotrophy in spontaneously hypertensive rats(SHRs).Briefly,safety and toxicity of BYL was evaluated by a toxicity experiment in Wistar rats.Thereafter,pharmacodynamics experiment was done in SHRs to test the effects of BYL on hypertension and cardiac hypotrophy.Combined with the RNASeq analysis and metabolomics analysis,mechanism of BYL on SHRs was determined.1.Safety evaluation of compound apitherapy preparation Bao-YuanLing:acute and sub-acute oral toxicity in Wistar ratsIn this study,acute and sub-acute toxicity test of Bao-Yuan-Ling(BYL)was done following Organization for Economic Co-operation and Development(OECD)423 and OECD 407 in Wistar rats,respectively.In acute toxicity test,rats were orally treated with BYL at the single dose of 2000mg/kg and 5000mg/kg.No death occurred in 7d acute toxicity test indicating that the lethal dose 50%value exceeded 5000 mg/kg.In subacute toxicity study,rats were treated with BYL at the dose of 250mg/kg,500mg/kg and 1000mg/kg in a daily base for continuous 28 days.Results showed that female rats were more likely affected by BYL in body weight changes,while biochemical indicators of blood serum in male rats were more susceptible to drug effects.However,neither female nor male rats had a limited influence on the organs in hematoxylin-eosin(HE)staining analysis.Accordingly,it was suggested that BYL was safe with a slightly toxic.2.Evaluation of the efficacy of phamacognosy of BYL in spontaneous hypertensive rats(SHRs)Ten male Wistar rats were set to be the Normal control group.Forty male SHRs were set to be Model control group,BYL-L group,BYL-M group and BYL-H group,which were administrated with BYL with 0,0.5,1.0,2.0 g/kg,respectively.Besides,another ten SHRs were set to be the Positive group with the treatment of valsartan orally.During the 5week treatment,blood pressure of SHR we measured with the result that BYL treatment could reduce the blood pressure of SHR.Thereafter,we found that BYL treatment reduced the serum level of Angiotensin II,endothelins 1 and transforming growth factor-β and improved the myocardial structure.Besides,the result of quantitative Real-time PCR indicated that B YL treatment could up-regulate the mRNA expression of PPAR-α and PPAR-y.Thus,we can conclude that BYL,the novel apitherapy formulation,had hypotensive and cardioprotective effect in SHR,and this effect might be achieved through the improvement of myocardial energy metabolism.3.RNA-Seq analysis of the effects of BYL in SHRsIn this study,we aimed to characterize the effects of BYL on cardiac hypotrophy in SHRs via RNA-Seq analysis.Consumption of 2.0 g/kg BYL for 5 weeks improved the cardiac status in SHRs.In heart tissues analyzed by RNA-Seq,1297 differentially expressed genes(DEGs)were detected.Of these DEGs,410 were enriched in KEGG pathways,including the insulin signaling pathway,phosphatidylinositol 3kinase/Akt signaling pathway,and mitogen-activated protein kinase signaling pathway.Moreover,overlapping DEGs in these pathways played important roles in the insulin signaling pathway.In summary,our results show that the effects of BYL on cardiac hypotrophy in SHRs were closely related to the insulin signaling pathway.4.Metabolomic study of BYL in SHRs using performance GC-TOF-MSIn this study,GC-TOF-MS diagram of serum samples from all the tested groups were tested.Thereafter,data were analyzed by SIMCA and the PCA scores plot indicated that the normal control(BC)group and BYL-H group met the significant degree of separation compared to the model control(MC)group,which was accordance with the result in the phamacognosy test that the BYL-H group had the most significant efficiency to the MC group.Compared to the OPLS-DA model,36 differential metabolites were recognized from the MC and BC group.The differential metabolites can mapped to mainly 11 pathways and regulate sugar metabolism,amino acid metabolism and lipid metabolism.Besides,BYL-H regulated the most differential metabolites back(5 differential metabolites)among the BYL treated groups.Differenctial metabolites between BYL-H and MC group could map to 5 pathways.Forty-eight signaling pathways containing 132 DEGs and 3 differential metabolites appeared to have key functions by the BYL-H administration,among which glycerophospholipd metabolism and glycine,serine,alanine and threonine metabolism played an important role in the BYL-H treatment.BYL-H treatment regulated the enzymes coded by Pla2g2d,Chka,Dgkg,and Cds2 and metabolites including creatine and D-glyceric acid.It was indicated that BYL-H improved the lipid metabolism and amino acid metabolism in SHRs.In summary,BYL was demonstrated to be a safe apitherapy formulation.Besides,BYL treatment can relieve the rising trend of blood pressure of SHRs and intervene with cardiac hypertrophy.According to the pharmacodynamic experiment,RNA-seq analyze and metabolomics analysis,the pharmaceutical effect was related to the energy metabolism. |
PDF Full Text Request