Study On Thermal Difference Yin Fu Pian And Yang Fu Pian Based Brown Adipose Tissue

At present,studies have found that brown adipose tissue(BAT)has become a new target for the treatment of metabolic diseases by promoting energy consumption and improving body metabolism.Brown adipose tissue is small in volume,but it has strong heat production capacity,which mainly depends on the specific protein in the tissue-UCP1.Proton leakage does not produce heat.This heat production mode is the main heat production mode of mammals.BAT is regulated by the hypothalamus of the central nervous system.It mainly regulates the activity of adenylate activated protein kinase(AMPK)in the hypothalamus,thus regulating the expression of corresponding nuclei or neuropeptides in the hypothalamus to control the central nervous system.Therefore,AMPK in the hypothalamus plays an important role in the regulation of energy metabolism.Radix Aconite,as the “essential medicine for warming yang”,can tonify the Yang Qi of all viscera.traditional Chinese medicine believes that Yang Qi is the part of the whole body with warm and exciting characteristics.therefore,from the perspective of energy metabolism,it can be interpreted as Radix Aconite can promote the energy metabolism of organisms and increase the heat production of organisms.A large number of studies show that warming drugs can act on brown adipose tissue,promote the expression of related thermogenic proteins,enhance energy metabolism and improve Yang deficiency symptoms.According to previous literature review,the current research report does not explore the thermal mechanism of Radix Aconite from the aspect of brown adipose tissue function.Yin Fu Pian and Yang Fu Pian,two unique processed products of Jianchang Bang,are also rarely studied at present.When the body has abnormal energy regulation,AMPK activity in hypothalamus will be regulated.BAT starts to work after receiving the signal from the center.On the one hand,BAT regulates the heat production of--UCP1,a specific protein in the cell,on the other hand,it controls lipid metabolism and maintains the energy balance of the body.Therefore,in this study,the effects of Yin Fu Pian and Yang Fu Pian on Yang-deficiency mice induced by rhubarb were observed from three aspects of hypothalamic AMPK protein-brown adipose tissue-lipid metabolism related signal pathway.among them,the molecular mechanism of Yin Fu Pian and Yang Fu Pian regulating brown adipose tissue and the mechanism of action affecting body metabolism and heat production were mainly explored from the perspective of Yin Fu Pian and Yang Fu Pian affecting brown adipose tissue function.On the other hand,it is compared with Shengfu Pian to explore whether the thermal properties of Radix Aconite processed into Yin Fu Pian and Yang Fu Pian have changed,and to observe whether the changes in comparative thermal properties are consistent with the difference between the different functions of Yin Fu Pian and Yang Fu Pian for men and women.The research on Jianchang Bang Yin Fu Pian and Yang Fu Pian is mainly improved from the above two aspects,so as to inherit and protect the traditional processing techniques of Radix Aconite,and provide a more complete theoretical basis for the application of Yin Fu Pian and Yang Fu Pian.The main research results are as follows:1 Study on Yin Fu Pian and Yang Fu Pian Regulating AMPK Expression in HypothalamusYang deficiency model was established by administration of rhubarb suspension in mice.After the model was successfully established,the liquid medicine was administered for 2 weeks.The expression of AMPK and corresponding genes in hypothalamus of mice were analyzed by Yin Fu Pian and Yang Fu Pian.The results showed that the effect on AMPK protein in hypothalamus: Compared with the normal group of the same sex,the expression of AMPK protein in female model group increased significantly.After administration of Yang Fu Pian,the protein level was inhibited and tended to return to normal state.The expression of AMPK protein in male groups was not significantly different.Effect on AMPK gene in hypothalamus:Compared with the same sex,the expression level of AMPK gene in model group andnormal group has statistical significance(P < 0.01).In female mice,the AMPK gene expression level in the Sheng Fu Pian group and the Yang Fu Pian group was significantly lower than that in the model group(P < 0.01).The administration group of male mice has significant difference compared with the same sex model group(P <0.01),of which Yang Fu Pian has the best inhibitory effect,followed by Sheng Fu Pian group.Effect on p-AMPK protein in hypothalamus of mice: In female mice,the expression of p-AMPK protein in model group is the highest,which is significantly different from that in normal group and Yang Fu Pian group(P < 0.05).For male mice,the expression result of p-AMPK protein is the same as that of female mice.Yang Fu Pian has the best inhibitory effect(P < 0.01),followed by Sheng Fu Pian group.2 Study on the Regulation of Brown Adipose Tissue Function by Yin Fu Pian and Yang Fu PianBy observing the expression of brown adipose tissue(BAT),cells,proteins and corresponding genes in rheum officinale-induced Yang deficiency model mice,this paper discusses how Yin Fu Pian and Yang Fu Pian regulate the function of brown adipose tissue.Yang deficiency model was established by administration of rhubarb suspension in mice.After the model was successfully established,brown adipose tissue was collected and hematoxylin-eosin(HE)staining was performed to observe BAT cell changes after 2 weeks of administration of liquid medicine.The expression of uncoupling protein 1(UCP1)and its m RNA were detected by Western blot and real-time fluorescence quantitative polymerase chain reaction(Real-time PCR).The results showed that The proportion of BAT:compared with the normal group of the same sex,the proportion of BAT in the model group decreased significantly(P < 0.01).Compared with the model group,the proportion of BAT in female mice Sheng Fu Pian and Yin Fu Pian increased significantly(P < 0.01),while there was no significant difference between each administration group and model group in male mice.The number of cells in BAT tissue: compared with the normal group of the same sex,the number of cells in female and male groups of the model significantly reduced(P < 0.05,P < 0.01);In female mice,the number of cells inYang Fu Pian group and Sheng Fu Pian group increased significantly compared with model group(P < 0.05),of which the Yang Fu Pian group performed better;For male mice,compared with the model group,the number of cells in Sheng Fu Pian group,Yin Fu Pian group and Yang Fu Pian group increased significantly(P < 0.01),of which Sheng Fu Pian group had the best effect.The expression of specific protein UCP1 in BAT: compared with the same sex,the expression level of UCP1 in model group and normal group is statistically significant(P < 0.05,P < 0.01).in female mice,the expression level of UCP1 in Yang Fu Pian group is significantly higher than that in model group(P < 0.05).each drug administration group of male mice is significantly different from that of model group of the same sex(P < 0.05),of which Yang Fu Pian is the most significant.The relative expression of UCP1 m RNA in BAT: compared with the normal group,the relative expression of UCP1 m RNA in the model group decreased significantly(P < 0.05,P < 0.01).in female mice,compared with the model group,the relative expression of UCP1 m RNA in Yang Fu Pian group,Sheng Fu Pian group and Yin Fu Pian group increased significantly(P < 0.05,P < 0.01).Compared with Yang Fu Pian group,the relative expression of UCP1 m RNA in Sheng Fu Pian and Yin Fu Pian was also significantly different(P < 0.05).In male mice,compared with the model group,The relative expression of UCP1 m RNA in Yang Fu Pian group was significantly different(P < 0.01),but there was no significant difference between Sheng Fu Pian and Yin Fu Pian.in addition,the relative expression of UCP1 m RNA in Sheng Fu Pian group and Yin Fu Pian group also has significant difference compared with Yang Fu Pian group(P < 0.05).3 Study on Regulation of Lipid Metabolism Pathway by Yin Fu Pian and Yang Fu PianPKA pathway is one of the important ways to regulate lipid metabolism.It can promote fat hydrolysis to glycerol and free fatty acids.The cyclic adenosine monophosphate(c AMP)and protease of PAK pathway were detected and analyzed by corresponding methods.The results show that the effect on c AMP in BAT of mice:For male mice,the c AMP content of model group is lower than that of normal group,while that of Yin Fu Pian and Yang Fu Pian is higher than that of model group,but thereis no significant difference.For female mice,the model group is significantly lower than that of the normal group,and the c AMP content in each group can be increased,among which the effect of crude aconite root is the best,followed by Yang Fu Pian.Effect on expression of PKA protein in BAT of mice: For female mice,there is no statistical significance between the groups,but the expression of PKA protein in the administration group has a certain upward trend,among which the Yang Fu Pian group has a better upward trend and is closer to the normal group.For male mice,compared with the model group,there are significant differences between the normal group and the Sheng Fu Pian group.The Yang Fu Pian and the Yin Fu Piant group have an upward trend,and the upward trend of the Yang Fu Pian group is closer to the Sheng Fu Pian group.Effect on PKA gene expression in BAT of mice: For female mice,compared with model group,there is a significant difference between normal group and Yang Fu Pian group,and the gene expression in Sheng Fu Pian group and Yin Fu Piant group increases.For male mice,compared with the model group,there are significant differences between the normal group and the administration group,and the expression level of Yang Fu Pian group is the highest.Effect on HSL protein expression in BAT of mice: For female mice,compared with the model group,there are significant differences between the normal group and the Yang Fu Pian group.The Sheng Fu Pian group and Yin Fu Pian group have an upward trend,and the Yin Fu Pian group has a higher upward trend than the Sheng Fu Pian group.For male mice,there may be large differences within the group,and there is no obvious difference between the male groups,but the expression of HSL protein in the administration group has a certain rise,with the rising trend of Sheng Fu Pian group and Yang Fu Pian group being better.Effect on HSL gene expression in BAT of mice: For female mice,compared with the model group,there are significant differences between the normal group and the administration group,and the gene expression in Yang Fu Pian group is the highest,which is consistent with HSL protein expression.For male mice,HSL protein does not show much difference,but gene expression has certain difference.Compared with the model group,there is a significant difference between the normal group and theadministration group.Yin Fu Pian group has the highest expression level,but the analysis of comprehensive protein expression results shows that Yang Fu Pian group can activate HSL protein more effectively.This study also used kits to detect triglyceride,low density lipoprotein and high density lipoprotein respectively.the analysis shows that: the determination of triglyceride: compared with model group,the triglyceride in Sheng Fu Pian group,Yin Fu Pian group and Tang Fu Pian group also decreased(P < 0.05);For males,Yang Fu Pian has the most obvious effect.For females,Yin Fu Pian has the best reducing effect.Determination of high density lipoprotein: the value of high density lipoprotein in mouse model group was significantly lower than that in normal group regardless of male and female(P < 0.05);Compared with the model group,the high density lipoprotein of Sheng Fu Pian group,Yin Fu Pian group and Yang Fu Pian group increased significantly(P < 0.05).There was no significant difference among the three groups.No matter male or female,the effect of radix aconiti lateralis preparata is the best,and the increase is the most obvious.Determination of low density lipoprotein: for females,compared with the model blank group,the low density lipoprotein of Yang Fu Pian group is significantly lower(P < 0.05);For males,there is no significant difference compared with normal group and model blank group.Conclusion: Yang deficiency mice have stress reaction,promote the expression of AMPK in hypothalamus,affect the thermogenic function of brown adipose tissue,and significantly reduce the lipid metabolism function of triglyceride.After administration,Yin Fu Pian and Yang Fu Pian exert thermogenic effect through hypothalamic AMPK protein-brown adipose tissue-lipid metabolism related signaling pathway.Their thermogenicity is still different,but the difference between male and female’s different functions has not been clearly reflected in this study.In general,both male and female,Yang Fu Pian and Yin Fu Pian have the effect of supplementing fire and yang,and Yang Fu Pian has better effect than Yin Fu Pian.therefore,it is suggested that Yang Fu Pian be used for patients with severe Yang deficiency and Yin Fu Pian for patients with mild yang deficiency or weak constitution in clinic.