Lentivirus-mediated overexpressed NR4A1 monocytes in the treatment of lupus nephritis in lupus mice

Abstract:

Objective: To explore whether overexpression of NR4A1 can alleviate lupus nephritis (LN). Methods: A mouse model of LN was constructed by intraperitoneal injection of 0.5 mL pristane, divided into saline control group (SC), pristane control group (PC), vector control group (VC) and overexpressed NR4A1 group. At the end of 20^thweek, the monocytes overexpressed NR4A1 were injected into the tail vein. The diameter of ankles was measured at the end of 28^thweek. Renal pathological sections were stained with HE staining and semi-quantitative scores were used to assess renal pathological damage. Serum TNF-α and anti-ds-DNA concentrations were measured by ELISA. The expression of Bcl-2 and NR4A1 in the spleen was detected by real-time PCR. Results: The results showed that compared with the SC group, LN-related activity indicators in the PC group were significantly increased, but the ankle joint diameter[4.69 (4.26, 5.13) mm, P=0.000], the anti-ds-DNA antibody concentration [(173.11±25.49) ng/mL, P=0.000], renal pathology score[2.78 (2.68, 2.81), P=0.000], and the expression of NR4A1 (0.40±0.09, P=0.008) was significantly reduced. Compared with the PC and VC groups, the NR4A1 group LN activity indicators were significantly improved, the ankle joint diameter[3.80 (3.44, 4.20) mm, P_PC=0.002, P_VC=0.033], the concentration of anti-ds-DNA antibody [(51.92±10.91) ng/mL, P_PC=0.008, P_VC=0.025], renal pathology score[1.39 (1.30, 1.49), P_PC=0.002, P_VC=0.018].ELISA results showed that the concentration of TNF-α in the NR4A1 group [(542.86±108.28) pg/mL]was significantly lower than that of the PC group [(1 779.00±358.40) pg/mL, P=0.008]and the VC group [(1 759.71±280.27) pg/mL, P=0.010].Real-time PCR results showed that the expression of Bcl-2 gene in the NR4A1 group (1.92±0.58) was significantly higher than that of the PC group (0.48±0.20, P=0.000). Conclusion: Overexpression of NR4A1 alleviates the symptoms of LN and reduces LN activity indicators in LN mice, which may involve the interaction among NR4A1, TNF-α and Bcl-2.