| BackgroundMicroRNA is a newly discovered non-coding RNA that can regulate the multi-protein genomics in human body.It has a highly conservative endogenous.After successful complementary and paired combination with the corresponding target gene at its site,it participates in the regulation of various physiological and pathological processes in the body by regulating the expression of target genes after transcription,and regulates the activation,proliferation,differentiation,apoptosis of cells,as well as the growth and development of organisms,playing a very important regulatory role in the immune response of the body,and the occurrence,development and outcome of diseases.At present,miRNA has become one of the hot spots in life science research.Studies have confirmed that serum miRNA expression in diabetic patients is dysfunctional.The dysfunctional miRNA acts on glucose metabolism process of the body through insulin sensitivity and resistance,islet β cell development,insulin production and secretion,and insulin signal transduction pathway,leading to the occurrence and development of insulin resistance and diabetes.At present,the standard treatment for patients with diabetes and end-stage renal disease is the combined pancreas and kidney transplantation,which can reduce the mortality related to cardiovascular diseases in patients.On this basis,it is of great significance to alleviate osteoporosis in middle-aged and elderly patients with chronic renal failure and delay the development of chronic complications of diabetes,and the quality of life of patients can be significantly improved.Diabetic nephropathy patients can return to normal blood glucose levels early after combined pancreas-kidney transplantation,in which miRNA plays an indispensable role.ObjectiveThe miRNA microarray was applied to screen and analyze the specific miRNA expression profiles related to the normalization of blood glucose before and after simultaneous pancreas and kidney transplantation in patients with diabetic nephropathy,to find the biomarkers of miRNAs that can predict the recovery of blood glucose after surgery and their regulated target genes,which can help to understand the physiological mechanisms of blood glucose regulation after simultaneous pancreas and kidney transplantation and provide new ideas for clinical diagnosis of patients with abnormal blood glucose after simultaneous pancreas and kidney transplantation.MethodsPatients with diabetic nephropathy who underwent combined pancreatic and renal transplantation in our hospital from January 2021 to December 2022 were selected as study subjects.The blood collection time was specified as 1 day before surgery and 3 months after surgery,and a total of 2 blood specimens were collected from each patient who met the criteria.Four patients were randomly selected from the patient who met the enrollment criteria using the digital table method,and the miRNA microarray technology was applied to detect the expression levels of serum miRNAs.q RT-PCR was used to verify the expression levels of the screened miRNAs,and target gene prediction and signaling pathway enrichment analysis were performed for the differential miRNAs,and subsequently the miRNA-regulated target genes associated with postoperative blood glucose recovery were screened.The target genes related to postoperative glucose recovery were screened,and the expression of target genes in serum was verified using q RT-PCR with Westernblot technique,and finally the correlation analysis between miRNAs and target genes was performed.ResultsThe expression of miR-320 d,miR-4701-3p,miR-937-5p,and miR-940 was significantly upregulated,and miR-1229-5p,miR-125a-3p,miR-3614-5p,miR-3656,miR-4459,miR-6090,in serum of patients with diabetic nephropathy after combined pancreatic and renal transplantation.miR-6769b-5p,miR-6800-5p,miR-6821-5p and miR-6893-5p expression were significantly down-regulated with statistically significant differences(P<0.05).By target gene prediction and enrichment analysis,four miRNAs(miR-125a-3p,miR-320 d,miR-940 and miR-1229-5p)were screened for possible involvement in the regulatory mechanism of glycemic recovery after combined pancreatic-renal transplantation in patients with diabetic nephropathy,and miR-320 d,miR-940 were found to be elevated in pancreatic tissues,miR We found that miR-320 d and miR-940 expression was elevated in pancreatic tissue,miR-1229-3p and miR-1229-5p expression was decreased,and through literature review,we found that the differential miRNA-regulated target genes IGFBP1 and THBS1 were involved in glycemic recovery.q RT-PCR and Westernblot detected their expression and found that IGFBP1 was elevated in diabetic nephropathy patients after combined pancreas-renal transplantation,while THBS1 was decreased.were statistically significant(P<0.05).Correlation analysis revealed that miR-940 was correlated with IGFBP1,which may be the target gene of miR-940,and miR-125a-3p was correlated with THBS1,which may be the target gene of miR-125a-3p.ConclusionPatients with diabetic nephropathy have specific miRNA expression spectrum before and postoperative serum.These differently expression miRNAs may be used as a biomagitory recovery of blood sugar recovery after the combined transplantation of diabetic nephropathy.It may be used as a regulatory factor to participate in the development of diabetic nephropathy. |
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