Probing the effect of SLIT3 on high glucose-induced inflammatory response in human glomerular podocytes based on JAK/STAT3 signaling pathway
Objective To investigate whether SLIT3 can modulate the JAK/STAT3 signaling pathway to reg-ulate high glucose-induced inflammatory responses in renal cellular podocytes.Methods Glomerular podocytes MPC-5 were randomly divided into L-Glucose+NC siRNA group,L-Glucose+SLIT3 group,L-Glucose+SLIT3 siRNA group,H-Glucose+NC siRNA group,H-Glucose+SLIT3 group,H-Glucose+SLIT3 siRNA group.Flow cytometry was used to detect the apoptosis rate;ELISA was used to detect the level of inflammatory factors in cells;protein blotting was used to detect the protein expression of JAK2,p-JAK2,STAT3,and p-STAT3 in cells;and RT-PCR was used to detect the relative expression of SLIT3,IL-1β,IL-6,and TNF-α in cells.Results Compared with the Ctrl group,SLIT3-siRNA1(0.67±0.04),SLIT3-siRNA2(0.26±0.03),and SLIT3-siRNA3(0.71±0.08)MPC-5 cells showed significantly lower expression of SLIT3 pairs,and the most significant change was observed in the SLIT3-siR-NA3 group(P＜0.01).Compared with the L-Glucose+NC siRNA group,the apoptosis rate of MPC-5 cells in the L-Glucose+SLIT3 group and the H-Glucose+NC siRNA group(11.84％±1.06％)(11.88％±1.07％),the cellular ex-pression of IL-1β(164.69±6.92)(155.31±8.62),IL-6(162.16±5.56)(105.35±3.44),TNF-α(103.31±6.84)(128.43±4.82)levels and relative mRNA expression in the cells,p-JAK2/JAK2(1.74±0.14)(3.93±0.17)and p-STAT3/STAT3 ratios(1.99±0.24)(4.43±0.16)were significantly increased,and the apoptosis rate(2.73％±0.25％),cellular levels of IL-1 β(78.09±7.47),IL-6(46.48±2.63),TNF-α(42.61±3.10),and the relative ex-pression of mRNAs in the L-Glucose+SLIT3 siRNA group,and cellular p-JAK2/JAK2(0.16±0.04)and p-STAT3/STAT3(0.20±0.03)ratios were significantly reduced(P＜0.01);compared with the H-Glucose+NC siRNA group,the apoptosis rate of MPC-5 cells in the H-Glucose+SLIT3 group(16.24％±1.28％),cellular IL-1 β(214.09±9.28),IL-6(205.44±6.80),TNF-α levels(192.11±4.93)and relative mRNA expression,and p-JAK2/JAK2(5.00±0.39)and p-STAT3/STAT3 ratios(5.71±0.33)in the cells were significantly increased in the H-Glucose+SLIT3 siRNA group apoptosis rate(6.25％±0.43％),cellular levels of IL-1β(118.60±8.05),IL-6(74.37±4.91),TNF-α(98.76±4.81),and relative expression of mRNAs,cellular p-JAK2/JAK2(2.99±0.13)and p STAT3/STAT3(2.05±0.14)ra-tios were significantly reduced(P＜0.01).Conclusion Knockdown of SLIT3 inhibits high glucose-induced inflam-matory response and apoptosis in glomerular podocytes and ameliorates podocyte injury,and its mechanism of action may be related to the inhibition of the activation of the JAK2/STAT3 signaling pathway.
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