Effect Of Bushen Yanling Prescription On Oxidative Stress And Inflammatory Indexes And MiRNA-mRNA Mechanism In Aging Population With Kidney Deficiency

Aging is an inevitable phenomenon when social economy develops to a certain stage.According to data released by the National Bureau of Statistics,by the end of 2018,China's population aged 60 or above had reached 249 million,accounting for 17.9%of the total population,an increase of 0.6 percentage points over 2017.Among them,the population aged 65 or above had reached 167 million,accounting for 11.9%of the total population,an increase of 0.5 percentage points over 2017.Presents the aging population base is big,fast growth,the serious situation of old before rich,premature aging,and aging population in our country's slow disease prevalence into a rising trend,disease burden is increasing year by year,China's 202 million elderly people at least more than 1 million people suffer from a chronic disease,and suffering from a variety of chronic diseases in the elderly has become increasingly serious.Aging is a gradual process,which is necessary for animals in nature from birth to death.It means the degeneration of body functions.Although aging is inevitable,it can be delayed.In the face of the approaching of aging society,it is the most fundamental way to let every citizen learn to keep in good health scientifically,delay aging,prevent diseases and avoid serious diseases.President Xi Jinping has pointed out that "without the health of the whole people,there can be no well-off society in all respects".We are faced with the common problem of delaying aging and living a long and healthy life.As China's traditional medicine,traditional Chinese medicine has accumulated rich experience in more than two thousand years of medical practice,has its own advantages in anti-aging,and has formed a unique method and theoretical system.Therefore,it is particularly important to explore the anti-aging theory and law prescriptions in traditional medicine to actively deal with social aging.ObjectiveTo sort out the understanding of the law and mechanism of aging in traditional Chinese medicine,summarize the rule of anti-aging prescription and the main categories of anti-aging traditional Chinese medicine.To investigate the effects of Bushen Yanling prescription on oxidative stress and inflammatory indexes in aging population with kidney deficiency,and to evaluate its safety;To explore the mechanism of kidney deficiency and senility and the mechanism of Bushen Yanling prescription.Methods1.Selected the representative ancient books of traditional Chinese medicine of each dynasty in the History of Chinese Medicine,and used "aging","endowment","old"and "old" as the key words.The Chinese Medical Book V5.0 software was used to conduct keyword retrieval for the above-mentioned included ancient books,and the retrieval results were exported into text document format.The retrieval results were sorted out,and the traditional Chinese medicine was classified according to the classification method of Chinese Medicine.2.Clinical study of 30 patients with kidney deficiency was conducted by using prospective self-controlled study before and after.The subjects were divided into pre-aging group and aging group.Mild kidney deficiency group,serious kidney deficiency group.Bushen Yanling prescription was used for intervention.After 8 weeks,the indexes of 30 patients before and after treatment were observed.The main therapeutic indexes included TCM syndrome score of senile kidney deficiency,Borg fatigue index scale,SOD,TNF-?,IL-6,testosterone,estradiol,complement C3 and complement C4 levels,and the safety indexes included liver function,renal function,blood routine,adverse events/reactions,etc.SPSS26.0 software was used for statistical analysis.Adverse reaction analysis was performed for those who withdrew from the study due to adverse events.Measurement data will use the mean±standard deviation according to statistical description of(X±S)count data with statistical description,frequency or percentage of paired sample t test made on the test results before and after,using independent sample t-test is compared between group,statistical test adopt bilateral test,P<0.05,think that the difference was statistically significant,The frequency was used for statistical description of the count data,and the chi square test was used for statistical description of the count data.Fisher's exact probability method was used for statistical description.3.Select aging from clinical trials of the subjects in the group,kidney empty is restructuring in 6 cases,6 cases with age in healthy subjects recruited the selection of subjects were 6 cases,the aging kidney were 6 cases,on the high flux in sequence,filtering through the analysis of the differences in expression of micrornas associated with kidney aging,the difference of mRNA expression patterns,and bioinformatics analysis,To explore the mechanism of senility of kidney deficiency.After taking the medicine for 4 weeks,the subjects in the clinical trial were subjected to high-throughput sequencing before and after taking the medicine,and the target genes of the effect of the prescription were screened through differential expression analysis.qt-PCR verification was conducted according to the clinical results in the second part to study the mechanism of the Bushen Yanling prescription.Results1.Summarized 59 signs or manifestations related to aging,and found that according to the theory of traditional Chinese medicine,the above-mentioned symptoms of aging were mainly related to kidney deficiency;To summarize the understanding of the mechanism of aging in traditional Chinese medicine,it is mainly Yin deficiency induced senility,Yang deficiency induced senility,kidney deficiency induced senility,Qi and blood deficiency induced senility,and evil Qi induced senility.To summarize the function or main indications of TCM anti-aging prescriptions,it is found that they are related to kidney deficiency,and the prescriptions of anti-aging prescriptions are still mainly kidney-tonifying drugs,qi tonifying drugs,blood tonifying drugs and tranquilizing drugs,supplemented by dispelling evil spirits.The anti-aging traditional Chinese medicines mainly fall into the category of tonifying deficiency and dispelling evil spirits,among which the traditional Chinese medicines for tonifying deficiency can be further divided into the category of tonifying qi,tonifying blood,tonifying Yin,tonifying Yang and tranquilizing the mind.Traditional Chinese medicine for removing pathogenic factors can be subdivided into heat-clearing,blood-promoting,phlegm-resolving,rheumatoid and water-enhancing medicines.2.In terms of TCM syndrome scores,after the comparison before and after treatment,the TCM syndrome scores of the subjects with kidney deficiency and aging decreased significantly after treatment,with statistical significance(P<0.001).After the treatment,the senility symptoms of the subjects with kidney deficiency were significantly improved,and the total effective rate of the treatment syndrome was 97%.According to age stratification,the syndrome scores of the pre-aging group were lower before and after treatment,suggesting that the symptoms of kidney deficiency in the pre-aging group were generally mild,but there was no statistical difference in the syndrome scores before treatment between the two groups(P>0.05).There was no statistical difference in syndrome scores after treatment(P>0.05),suggesting that Bushen Yanling prescription was consistent in improving symptoms of kidney deficiency and aging in patients of different ages,and there was no difference in efficacy.In terms of physical improvement,Borg fatigue index scores of subjects were significantly decreased after treatment after comparison,with statistical significance(P<0.001).The fatigue of the subjects was improved after treatment.In the aspect of antioxidant function,SOD level was significantly increased after treatment(P<0.05);In terms of reducing inflammatory response,the level of IL-6 decreased after treatment,but the difference was not statistically significant(P>0.05).The level of TNF-? decreased significantly after treatment(P<0.05).As for other anti-aging indexes,the level of sex hormone secretion after treatment had a tendency to increase,but there was no statistical significance(P>0.05).After treatment,the levels of complement C3 and complement C4 were basically the same as before treatment(P>0.05).According to the age stratification,there was no difference between the groups before the treatment of each item,and there were differences between the groups after the treatment of SOD(P<0.05).The improvement of SOD was more obvious in the pre-aging group,suggesting that the application of Bushen Yanling prescription in the pre-aging period(40-50 years old)had a more significant effect of anti-oxidative stress.There was no difference in the other items between the groups after treatment(P>0.05).In terms of the results of the intervention of inflammatory factors,sex hormones and complement,the efficacy of different ages was consistent.IL-6 and TNF-? were found to be different before treatment by symptom stratification.IL-6 was higher in mild renal deficiency group and lower in renal deficiency group than in recombination group.TNF-? was lower in mild renal deficiency group,and higher in renal deficiency group than in recombination group.The difference disappeared after treatment.There was no difference between the other items after treatment(P>0.05),and the efficacy of SOD,inflammatory factors,sex hormones and complement was consistent.During the study period,no serious adverse events occurred in the subjects,and AST decreased significantly after treatment(P<0.05).CR increased after treatment,with statistical difference compared with before treatment(P>0.05),but before and after treatment,blood routine,liver function,renal function of subjects before and after treatment did not show significant abnormalities,the results showed that Bushen Yanling prescription is safe and reliable.3.Comparing the differences between aging and non-aging,there were 272 different mRNAs with a difference ratio of FC>1.5 and P<0.05,among which 137 mRNA were up-regulated and 135 mRNA were down-regulated.There were 51 differential miRNAs,including 24 up-regulated miRNAs and 27 down-regulated miRNAs.The difference between kidney deficiency and non-kidney deficiency was 174 mRNA with FC>1.5 and P<0.05,among which 100 mRNA were up-regulated and 74 mRNA were down-regulated.There were 149 differential miRNAs,of which 105 were up-regulated and 44 were down-regulated.By intersecting the different mRNAs of the two groups,8 mRNAs related to age and kidney deficiency syndrome could be obtained,which were H3C6,ARL17B,FOXD2,DNAH14,ighv3-43,SLC12A1,TBC1D3I,and AC105001.2.By intersecting the different miRNAs of the two groups,8 miRNAs related to both age and kidney deficiency syndrome could be obtained.These were miR-11987-z,miR-13-y,bantam-y,miR-317-z,miR-137-y,miR-276-y,miR-311-z,and hsa-miR-1246.Pair comparison before and after treatment showed 221 different mRNAs,including 71 up-regulated mRNAs and 150 down-regulated mRNAs.There were 152 differential miRNAs,among which 46 were up-regulated and 106 were down-regulated.25 potential age-related mRNA targets of Bushen Yanling prescription:COL3A1,COL1A2,POSTN,ELN,CDH11,ABI3BP,PENK,COL12A1,BGN,PRRX1,ITGA11,CLDN10,COL1A1,GJA1,DCN,IL6,COL5A1,SPDYC,PAPPA,COL6A3,IGFBP4,FOXD2,ADAM12,GCNT7,CHGA,8 potential miRNA targets were bantam-Y,miR-13-y,miR-137-y,miR-11987-z,miR-317-z,miR-276-y,novel-m0145-5p,and hsa-miR-1246.9 mRNA targets of SLX1B,ARG2,PROS1,KCND3,FOXD2,FAXDC2,VSIG2,MFSD6L,TEDDM1,which were potentially related to kidney deficiency syndrome were obtained.19 potential miRNA targets were bantam-y,miR-13-y,miR-137-y,miR-11987-z,miR-317-z,miR-276-y,novic-m0212-3p,hsa-miR-1197,novic-m0004-5p,novel-m0005-5p,novel-m0009-3p,miR-584-x,miR-144-y,hsa-miR-375-3p,miR-313-y,hsa-miR-183-3p,novel-m0175-3p,miR-3355-z,hsa-miR-1246.1 potential mRNA target related to both age and kidney deficiency syndrome:FOXD2;6 miRNA targets:bantam-y,miR-137-y,miR-11987-z,miR-317-z,miR-276-y,and hsa-miR-1246.The regulation of the Bushen Yanling prescription on the target gene is that hsa-miR-371b-5p is up-regulated,IL-6 is down-regulated,GFAP is down-regulated,and p21 is down-regulated.COL6A3 is down-regulated,PI3K is down-regulated,AKT is down-regulated,FOXO is up-regulated,SOD2 is up-regulated.Conclusion1.Kidney deficiency is the "internal cause",exogenous pathogenic factors is the"external cause",and the aging process is accompanied by the decline of qi and blood.Kidney deficiency is the main factor of senility and the common witness of senility.The ancient antisenility prescription consists of kidney tonifying medicine,qi tonifying medicine,blood tonifying medicine and tranquilizing medicine,supplemented with dispelling evil spirits.Anti-aging traditional Chinese medicines include Qi tonifying,blood tonifying,Yin tonifying,Yang tonifying,calming the mind,clearing heat,promoting blood circulation,resolving phlegm,expelling rheumatism and improving water.2.Bushen Yanling prescription can improve the symptoms related to kidney deficiency and aging,improve the level of SOD in people with kidney deficiency and aging,reduce the level of TNF-?,improve the anti-oxidative stress function of human body,and reduce the level of inflammatory response,which is safe and effective.3.Bushen Yanling prescription may play an anti-oxidative stress role by down-regulating COL6A3,down-regulating PI3K,down-regulating AKT,up-regulating FOXO,up-regulating SOD2,improving SOD level;By up-regulating hsa-miR-371b-5p,regulating the down-regulation of IL-6,regulating the down-regulation of GFAP,and regulating the down-regulation of p21,the level of inflammatory response can be reduced.