Abstract
Systemic lupus erythematosus (SLE) is a common but severe autoimmune systemic inflammatory disease. Lupus nephritis (LN) is a serious complication of SLE,affecting up to 70% of SLE patients. Circulating microRNAs (miRNA) are emerging as biomarkers for pathological conditions and play significant roles in intercellular communication. In present research, serum samples from healthy control, early and late stage LN patients were used to analyze the expression profile of miRNAs by microarray. Subsequent study demonstrated that miR-130b-3p in serum of patients with early stage LN were significantly up-regulated when compared with healthy controls. In addition,we have also observed that the expression of a large amount of circulating microRNAs significantly decreased in patients with late stage LN. The further analysis found that the expression of serum miR-130b-3p was positively correlated with 24-hour proteinuria and renal chronicity index in patients with early stage LN.Transfection of renal tubular cellline(HK-2)with miR-130b-3p mimics can promote epithelial-mesenchymal transition (EMT). The opposite effects were observed when transfected with miR-130b-3p inhibitors. MiR-130b-3p negatively regulated ERBB2IP expression by directly targeting the 3′-UTR of ERBB2IP The circulating miR-130b-3p might serve as a biomarker and play an important role in renal damage in early stage LN patients.
Highlights
MicroRNAs are short non-coding RNA molecules that inhibit gene expression through incomplete base pairing with the 3′ -untranslated region (3′ -UTR) of target mRNAs7,8
We attempted to reveal the changes of miRNAs in serum during different stages of CKD caused by Lupus nephritis (LN) utilizing microarray technology, and investigated their correlation with renal damage and the possible molecular mechanism involved in early stage LN
We found that the expressions of seven miRNAs, including miR-130b-3p, miR-1233-3p, miR-18a-3p, miR-628-3p, miR-1260b, miR-1539 and miR-378e were increased in early stage LN patients compared to healthy controls (P < 0 .05) (Fig. 1A,B)
Summary
A total of 96 serum samples, including 60 LN patients with different stages of CKD and 36 healthy controls, were included in this study. Blood samples from 4 early stage LN patients(CKD 1–3 stage), 4 late stage LNpatients(CKD 4–5 stage) and 4 healthy controls were used to analyze the expression profile of circulating miRNA. RNA was reverse transcribed to cDNA using miScript Reverse Transcription kit (Qiagen,USA) according to the manufacturer’s instructions. Another 52 LN patients (including 40 patients with early stage LN and 12 patients with early stage LN), and 32 age- and sex-matched healthy control were recruited for the validation group. Seventy-two hours after transfection with miR-130b-3p mimics or inhibitors, the cells were lysed with RIPA buffer (Thermo Fisher Scientific,USA) respectively, and Western blotting was performed using standard procedures. All statistical analyses were performed and graphs were generated using GraphPad Prism 5.0 (GraphPad Software Inc, California)
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