Observation Of Therapeutic Effect And Transcriptome Analysis Of Congrongjing Decotion In The Treatment Of Parkinson's Disease

Objective:To observe the safety and therapeutic effect of medicine added Congrongjing Decotion(CRJD)through a randomized double-blind placebo controlled trial in the treatment of Parkinson's disease with the deficiency of kidney essence.Blood samples were randomly selected from patients before and after the trial to extract the mRNA.The RNA-seq technique was used to detect the mRNA.RT-PCR was used to verify the screening genes,so as to explore the mechanism of the effect of CRJD added medicine in the treatment of PD patients.Methods:86 patients who met the criterion were randomly divided into control and treatment groups,with 43 patients in each group.The treatment group was treated with CRJD and medicine,and the control group was treated with placebo and medicine.Two courses of treatment were 12 weeks.The total scores of the Unified Parkinson Disease Rating Scale(UPDRS)and its sub-items ?,?,?,? integrals,Parkinson Disease Question-39(PDQ-39 scores),Traditional Chinese Medicine(TCM)syndrome scores,medicine dosages,detect hematuria,liver and kidney function were recorded in before and after treatment to observe and compare the safety and therapeutic effect of CRJD.Blood samples of two groups of each 6 cases were randomly selected from patients before and after the trial to extract the mRNA.3 samples from each of the two groups which completed the clinical trials and RNA quality inspection qualified were used to construct a cDNA library for RNA-sequencing(RNA-seq).Six pairs of blood transcripts were obtained by RNA-seq technique.And the differentially expressed genes were screened out before and after the experiment.The functional cluster of differentially expressed genes was analyzed by using GO(Gene Ontology)method in combination with the clinical trial results.KEGG(Kyoto Encyclopedia of Genes and Genomes)method was used to analyze the signal pathway related to differentially expressed genes.And select the important differentially expressed genes acting on PD by adding and subtracting CRJD.Results:?Finally,72 cases were completed according to the protocol.37 cases in control group and 35 cases in treatment group.?Before and after the treatment,there was a significant difference between the scores of the UPDRS total score and sub-II(activities of daily living),the PDQ-39 scores,TCM syndrome scores,doses of LED(p<0.05).Comparison between groups after treatment,UPDRS sub-II,PDQ-39 score,symptom scores,medicine doses were statistically significant(P<0.05).?During the treatment period,there were 2 cases of adverse reactions which eased to disappeal after increasing the dosage of Salvia Miltiorrhiza and Radix Paeoniae or withdrawing drug.Compared with pre-treatment,the changes of safty indexes were not found to be significantly(P>0.05)of all patients.? GO analysis of differential gene:differentially expressed genes before and after trial in treatment groups(TH vs TQ)were divided into 35 functional clusters.Differentially expressed genes after trial in treatment and control groups(TH vs CH)were divided into 47 functional clusters.As a result,it was found that the differential genes are mainly concentrated in the process of cell regulation and metabolism in the biological process;the cell membrane and synapse of the cellular components.?Through the KEGG,it was found that PD-related differentially expressed genes are likely to be enriched in the following 4 signaling pathways:Ribosomal receptor interaction signaling pathway;Endocytosis signaling pathway;FOXO signaling pathway;Apoptosis-related signaling pathway.?According to the quantitative data of differential gene,there were 76 differentially expressed genes screened out in the treatment group before and after trial(TH VS TQ);Before and afeter the trial in control group(CH VS CQ),331 genes were screened differentially;Before and after the trial in two groups(TH VS TQ vs.CH VS CQ),a total of 13 genes were mutated together.?According to the results of RT-PCR verification of differential gene expression and literature analysis,NFE2L2,BCL2A1,and GCHl genes are associated with the CRJD for PD.Conclusion:? The safety and therapeutic effect ofmedicine added CRJD in the treatment of PD with the deficiency of kidney essence is well.It can reduce the dosage of medicine,improve the clinical symptoms of patients and improve their quality of life.? The results of functional analysis and the transcripts in the treatment of PD patients treated by CRJD were obtained by RNA-seq technique.? The related loci of CRJD acting on PD patients were screened out.