Abstract:Objective: To investigate the effects of microRNA-93-5p (miR-93-5p) on renal mesangial cell (RMC) damage in lupus nephritis (LN) mice. Methods: QRT-PCR was used to detect the expression of miR-93-5p and ROCK2 mRNA in the serum of 33 LN patients (LN group) treated in our hospital from July 2023 to May 2024 and 33 pairs of healthy individuals (control group) undergoing physical examination. RMCs (identified by morphology) were isolated and purified from LN mice and assigned into Control group, NC-mimics group (transfected with NC-mimics), miR-93-5p-mimics group (transfected with miR-93-5p-mimics), miR-93-5p-mimics+OE-NC group (transfected with miR-93-5p-mimics+OE-N), and miR-93-5p-mimics+OE-ROCK2 group (transfected with miR-93-5p-mimics+OE-ROCK2). The expression of miR-93-5p and ROCK2 mRNA was detected in the cell lysates of each group. MTT and flow cytometry were used to detect the effects of overexpression of miR-93-5p on RMC proliferation and apoptosis, respectively. ELISA kit was used to detect the expression of TNF-α, IL-6, and IL-1β in in the supernatant of RMC cells. In addition, Western blot was used to detect the expression of PCNA, Bcl-2, Cleaved Caspase-3, and ROCK proteins in the lysate of RMC cells. Results: Compared with control group, LN group showed a decrease in miR-93-5p (t=15.977,P<0.001) and an increase in ROCK2 (t=11.292,P<0.001). Compared with the Control group and the NC-mimics group, the values of A570, the expressions of miR-93-5p, PCNA and Bcl-2 in the miR-93-5p-mimics group were increased. The expressions of apoptosis rate, ROCK2 mRNA and protein, TNF-α, IL-6, IL-1β, and Cleaved Caspase-3 decreased (P<0.05). Compared with miR-93-5p-mimics group and miR-93-5p-mimics+OE-NC group, the miR-93-5p-mimics+OE-ROCK2 group showed a decrease in A570 value, miR-93-5p, PCNA, and Bcl-2, and an increase in apoptosis rate, ROCK2 mRNA and protein, TNF-α, IL-6, IL-1β, and Cleaved Caspase-3 expression (P<0.05). Conclusion: miR-93-5p inhibits RMC damage in LN mice by targeting ROCK2.
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