| Backgrounds:Sargassum(HZ)and Glycyrrhiza uralensis(GC)are anti-drug combinations.There are many records in the literature that are not compatible with anti-drug combinations and a few reports on adverse reactions in modern clinical practice.However,many doctors who used them in ancient and modern clinical practice also received good treatment effects.The Haizao Yuhu Decoction(HYD),containing HZ and GC anti-drug combination is a common prescription for clinical treatment of various diseases.As people's awareness of drug safety continues to increase,the question about the anti-drug combination has caused a lot of thinking and controversy,and many experimental studies have been carried out.Both HZ and GC are non-toxic Chinese medicines,and the nature of their incompatibility is inevitable.In summary of the previous experimental studies,we can find that there are many studies about the single-drug and anti-drug combination,but the research on the commonly used HYD is less,and the clinical use of compound decoction has achieved good results;there are many studies under physiological conditions,and the clinical application of anti-drug combination is basically under pathological conditions;both Haihaozi and Yangqicai can be used as medicines,and modern pharmacological experiments also indicate the chemical composition and the pharmacological activities of them are not the same,but the species of HZ used in the research is rarely mentioned,and there are few comparative study of different species.Therefore,in the early stage of the research,the amount-toxicity-effect relationship and mechanism of different species of HZ(Haihaozi and Yangqicai)and GC in HYD were studied under the pathological model of goiter.On the basis of above,the dose of HZ and GC in the study referring to the limit of 5 times of them recorded in the 2015 version of the "Chinese People's Republic of China Pharmacopoeia"(hereinafter referred to as"Chinese Pharmacopoeia"),we continue to studying the characterization and mechanism of nephrotoxicity in goiter rats causing by the addition and subtraction application of different species of HZ and GC in HYD.Objects:Based on the previous research,we explored the effects of different species of HZ and GC using the high dose limit of 5 times in the Chinese Pharmacopoeia when they were on the addition and subtraction of HYD,and the effects on the characterization and mechanism of heart,liver,kidney,blood cell parameters and serum oxidative antioxidant system toxicity.To explore whether its occurrence is related to different species of HZ,whether it is related to the application of anti-drug combination in compound,so as to better explore the essence of the contraindication theory of Chinese medicine,and provide scientific basis for clinical safe and rational drug use.Methods:1 Toxicity characterization:On the basis of the previous research of the research group,the doses of HZ and GC were selected five times higher than the dose limit specified in the Chinese Pharmacopoeia.Rats were randomly divided into blank group,model group,positive drug group,Haizao Yuhu Decoction(Haihaozi)without Glycyrrhiza uralensis group(HYDHaibaozi-GC),Haizao Yuhu Decoction(Yangqicai)without Glycyrrhiza uralensis group(HYDYangqicai-GC),Haizao Yuhu Decoction without HZ group(HYD-HZ),Haizao Yuhu Decoction(Haihaozi)group(HYDHaihaozi),Haizao Yuhu Decoction(Yangqicai)group(HYDyangqicai),and Haizao Yuhu Decoction without HZ and GC group(HYD-HZ-GC),except for blank group Propylthiouracil(PTU)replicated the goiter model and euthyroxine was used as a positive control.Serum liver function,renal function,cardiac function,serum oxidative and antioxidant related indicators,blood cell parameters and pathological changes of heart,liver and kidney tissues were observed.The doses of HZ and GC were selected five times higher than the dose limit specified in the Chinese Pharmacopoeia.The effect of addition and subtraction of different species of HZ and GC in HYD on the characterization of heart,liver,kidney,blood cell parameters and serum oxidative antioxidant system toxicity was observed.2Mechanism discussion:To detect the expression of CYP2E1,CYP3A1 mRNA and CYP2E1 protein in liver tissue,to explore the effect of hepatic enzyme activity on drug metabolism;to detect the expression of Nrf2 and HO-1 mRNA in liver tissue,and to explore the effect of oxidative stress on liver tissue Mechanism;detection of renal PGC-1? mRNA expression,to explore the mechanism of action of mitochondrial dysfunction on kidney tissueResults:Effects of high-dose addition and subtraction of different species of HZ and GC in HYD on the characterization toxicity in goiter rats.1.1 Liver:Compared with the blank group,the liver coefficient of the model group,serum ALT,AST,TP,ALB,GLB showed no significant changes.Under the light microscope,the liver tissue structure was normal;The ALT,AST,TP,ALB,GLB increased in HYDYangqicai-GC,HYD-HZ,HYDYangqicai,HYD-HZ-GC,and fat vacuoles were occasionally seen in the liver cells;No obvious changes were observed in HYDHainaozi-GC and HYDHaihaozi,under the light microscope,the liver tissue structure was normal.When combined with GC,Haihaozi and Yangqicai showed different trends in the liver,but there was no significant difference between them.1.2Kidney:Compared with the blank group,the renal coefficient of the model group,serum BUN,Cr,UA,Na+,K+,urine ?2-microglobulin,urinary EGF,urinary LDH were not significantly different,under light microscope,renal cortex and medulla show no obvious abnormalities;while the HYDyangqicai-GC,HYDyangqi,ai and HYD-HZ-GC increased the serum BUN and Cr,and under the light microscope,the tubular epithelial cells were swollen and vacuolar degeneration,and the glomerular shrunken;The glomerular capsule was shrunk,and K+,?2-microglobulin and urinary EGF were elevated in the HYD-HZ-GC,and no obvious abnormalities were observed in the remaining administration groups.1.3Heart:Compared with the blank group,the positive group and the model group,the heart coefficient of each group was decreased.Compared with the blank group,the CK and the LDH of HYD-HZ and HYDyangqicai were not changed,there was no significant change in CK and LDH in remain drug-administered group.No obvious damage was observed in the heart tissue of each drug-administered group under light microscope.1.4 Blood cell parameters:Compared with the blank group,there was no significant difference in RBC,HGB,HCT,WBC and PLT between the model group and each drug-administered group.The MCV,MCH and MCHC of the HYDHaihaozi and HYDyangqicai were normal,while the MCV and MCH of HYDuaihaozi-GC,HYDYangqicai-GC,HYD-HZ and HYD-HZ-GC were reduced.1.5 Serum oxidative antioxidant system:Compared with the blank group,there was no significant change in SOD in each drug-administered group.There was no significant difference in GSH-Px,XOD and H2O2 in the model group.GSH-Px,XOD and H2O2 in the HYDyangqicai were increased.The GSH-Px and H2O2in the HYDYangqicai-GC increased,and the PTU caused the MDA of the model group to increase.Except that the MDA of the HYDHaihaozi-GC returned to normal,the MDA content of the remaining drug-administered groups was no different from the model group.2Effects of high-dose addition and subtraction of different species of HZ and GC in HYD on the mechanism of toxicity in goiter rats.2.1Effects on liver CYP450 enzyme:Compared with the blank group,PTU significantly induced the expression of CYP2E1 mRNA in the model group,and each administration group corrected the overexpression of CYP2E1 mRNA.There was no significant effect of CYP3A1 mRNA and CYP2E1 protein expression in each drug-administered group.2.2 Effects on liver Nrf2/HO-1 pathway:Compared with the blank group,PTU significantly inhibited the expression of Nrf2mRNA in the model group.The expression of Nrf2 mRNA in the HYDHaihaozi-GC and HYDHaihaozi returned to normal,and the remaining groups were not different from the model group.PTU had no effect on the expression of HO-1 mRNA.The expression of HO-1 mRNA was significantly induced in HYDHaihaozi-GC and HYDHaihaozi,and there was no significant difference in the remaining.2.3The effect on the expression of PGC-1? mRNA and protein in the kidney:There was no significant difference in the expression of PGC-1? mRNA in all groups,among which HYDHaihaozi-GC had a tendency to induce the expression of PGC-1 a mRNA in kidney,while the expression of PGC-1? mRNA in the remaining groups had a tendency to decrease;The expression of PGC-1? protein had no significant effect in all groups.Conclusion:1 The different species of HZ and GC using the high dose limit of 5 times in the Chinese Pharmacopoeia when they were on the addition and subtraction of HYD,the liver function of HYDYangqicai-GC,HYD-HZ,HYDYangqicai and HYD-HZ-GC was affected,and the protein synthesis function of the liver was promoted;PTU induced the production of MDA,and the deregulated oxidative and antioxidant function of HYDYangqicai-GC and HYDYangqicai causing the Nrf2/HO-1 Pathway activation disorder,while HYDHaihaozi-GC and HYDHaihaozi increased the antioxidant capacity by promoting the expression of Nrf2 and HO-1 mRNA,which can protect the liver tissue from peroxide damage to some extent.When applied in HYD,Haihaozi and Yangqicai showed different trends in the liver,but there was no significant difference between them.2 The different species of HZ and GC using the high dose limit of 5 times in the Chinese Pharmacopoeia when they were on the addition and subtraction of HYD,PTU induced the increase of CYP2E1 mRNA in the model group,the overexpression of CYP2E1 mRNA was corrected in each administration group,but there were no significant differences between them.Each administration group had no significant effect on the expression of CYP3A1 mRNA and CYP2E1 protein,The compatibility of different species of HZ with Glycyrrhiza uralensis may not produce toxicity enhancement or attenuation by regulating the expression of CYP3A1 gene.In the later stage,the enzymatic activity and gene and protein expression of CYP2E1 and CYP3A1 should be further discussed..3The different species of HZ and GC using the high dose limit of 5 times in the Chinese Pharmacopoeia when they were on the addition and subtraction of HYD,the renal function of HYDYangqicai-GC,HYDYangqicai and HYD-HZ-GCwas affected,and no obvious abnormalities were found in the remaining groups,the effects of using Yangqicai and Haihaozi in HYD on kidney tissue were different.The expression of PGC-la mRNA and protein in the kidneys showed no difference in all groups.It is speculated that the effects of different species of HZ and GC on kidney tissue when they were on the addition and subtraction of HYD may not be regulated by regulating the expression of PGC-1alpha gene synthase to affect mitochondrial function.Each drug group had no significant effect on cardiac tissue and blood system.There was no significant difference between different species of HZ. |
PDF Full Text Request