| Objective: Alpinia Oxyphylla is the dried and mature fruit of Alpinia Zingiberaceae,w hich is a common Chinese herbs tonifying the kidney in clinical.Traditional medicine believes that “the crude Alpinia Oxyphylla Fructus enter the spleen,but the mix-fry with brine one enter the kindey”,and its salt-processed products are widely used treatment of Kidney-yang deficiency in clinical.Whether the chemical composition changes after salt processing of Alpinia Oxyphylla? Is there a better effect on kidney disease after salt-processing of Alpinia Oxyphylla? Based on the theory of "intestine-kidney" interaction,what is the effect of salt processing of Alpinia Oxyphylla on intestinal flora? These scientific problems have become an important issue for indepth understand the clinical efficacy of the salt processing of Alpinia Oxyphylla.Therefore,this article intends to study the content difference of nine chemical components between crude Alpinia Oxyphylla Fructus(CAOF)and salt-processed Alpinia Oxyphylla Fructus(SAOF),and analyze the changes of the chemical components between CAOF and SAOF extract.The pharmacodynamics and intestinal flora of CAOF and SAOF were compared and analyzed by using diabetic nephropathy(DN)rat model and intestinal flora analysis technique,in order to reveal the scientific connotation of "SAOF enter the kidney".Methods: Ultra-efficient liquid-mass spectrometry(UPLC/MS/MS)was used to quantitatively analyze the contents of 9 chemical components in CAOF and SAOF;A single intraperitoneal injection of streptozotocin(STZ)solution 50 mg/kg to induced a rats model of DN,the renal related functional parameters of rats were investigated: total cholesterol(TC),Blood urine nitrogen(Bun),triglyceride(TG),creatinine(Scr),high density lipoprotein(HDL-C)and low density lipoprotein(LDL-C),the contents of TGF-β1 and α-SMA protein in rats kidney were determined by western blot method.The intestinal flora was sequenced by 16 S r RNA sequencing method,and the contents of acetic acid,propionic acid and butyric acid in fecal short chain fatty acids(SCFAs)of rats were detected by GC.Results: 9 compounds were determined by UPLC-MS/MS analysis,which included 1 terpenoid,5 flavonoids and 3 diarylheptanes.The content of each component in CAOF was higher than that in SAOF,and the contents of Nocardione,Oxyphyllacinol and Yakuchinone A were the highest.The levels of fasting blood glucose,24 h urinary protein,Scr,Bun,TG,TC and LDL-C were significantly increased,while the body weight and HDL-C were decreased in DN rats.The body weight,fasting blood glucose,24 h urinary protein and renal function parameters of rats in the treatment group were recovered after the intervention of CAOF and SAOF ethanol extracts,and the effect of SAOF was the same as that of CAOF;CAOF and SAOF ethanol extracts significantly decreased TGF-β1 and α-SMA in model group rats.In addition,CAOF and SAOF could significantly reduce the level of Enterobacteriaceae in the DN rats,and the levels of acetic acid,propionic acid and butyric acid in SCFAs and the abundance of Romboutsia and Prevotellaceae were also increased.Conclusion:1.Compared with CAOF,the contents of terpenoids,flavonoids and diarylheptane were decreased by 82.33%,85.29% and 89.37%,respectively.the content of terpenoid nocaketone has the highest,and the content of diarylheptane Yakuchinone A has the second highest.Salt-processing is a heating procedure,which increases the elimination of some groups,and result in changes in the content of chemical composition.2.Both CAOF and SAOF had protective effects on chronic kidney injury,but there was no significant difference between the two groups.Both CAOF and SAOF can improve the blood glucose,body weight and renal function parameters of DN rats,and regulate the proteins expression of TGF-β1 and α-SMA.It is speculated that the pharmacodynamic effect of Alpinia Oxyphylla is related to its regulation of the expression of related fibrogenic factors and improvement of inflammatory response in DN rats,so as to achieve the research on the protective effect of Alpinia Oxyphylla on renal injury in DN rats.3.The content of acetic acid,propionic acid and butyric acid in the intestinal flora metabolites SCFAs is significantly different.The content of SCFAs in the SAOF group is higher than that of the CAOF.The content of SCFAs in descending order were as follows: acetic acid,butyric acid,and propionic acid.CAOF and SAOF show a better regulation of the intestinal microbial function of DN rats by down-regulating the species abundance of Proteobacteria,Enterobacteriaceae and up-regulating the of prevotellaceae and Romboutsia.Through the above analysis,we found there are significant differences in the chemical composition content of SAOF and CAOF and the content of acetic acid,propionic acid and butyric acid in SCFAs.However,there is little difference in the pharmacodynamics and intestinal flora abundance in the DN rat model of the CAOF and SAOF ethanol extracts.For the later experiments,we will further study the effects of pharmacodynamic “salt processed enter the kidney” of Alpinia Oxyphylla. |
PDF Full Text Request