Volume 43 Issue 8
Jul.  2022
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Tongxin YANG, Guang WANG, Rui XU, Siwei SU, Kewei FANG, Jianhe LIU, Jiongming LI. Effect of Cathelicidin LL-37 on Transmembrane Barrier Function of Urothelial Cells[J]. Journal of Kunming Medical University, 2022, 43(8): 34-40. doi: 10.12259/j.issn.2095-610X.S20220806
Citation: Tongxin YANG, Guang WANG, Rui XU, Siwei SU, Kewei FANG, Jianhe LIU, Jiongming LI. Effect of Cathelicidin LL-37 on Transmembrane Barrier Function of Urothelial Cells[J]. Journal of Kunming Medical University, 2022, 43(8): 34-40. doi: 10.12259/j.issn.2095-610X.S20220806

Effect of Cathelicidin LL-37 on Transmembrane Barrier Function of Urothelial Cells

doi: 10.12259/j.issn.2095-610X.S20220806
  • Received Date: 2022-04-20
    Available Online: 2022-07-23
  • Publish Date: 2022-07-28
  •   Objective  To explore the specific effect of LL-37 on the destruction of the transmembrane barrier function of bladder urothelial cells and develop a cellular experimental model suitable for studying interstitial cystitis (IC) in vitro.   Methods   Human urothelial immortalized cells SV-HUC-1 were treated with LL-37 at different concentrations. The transepithelial electrical resistance (TEER) was measured by a transepithelial resistor, and the cell proliferative activity was detected by CCK-8 kit. Cell cycle and calcium ion concentration were detected by flow cytometry. The expression levels of heparan sulfate, a key component of glycosaminoglycan (GAGs), were detected by RT-qPCR and WB.   Results   In vitro measurement showed that 100 and 200 μg/mL of LL-37 significantly inhibited the TEER and destroyed the transmembrane barrier function of SV-HUC-1. The CCK-8 test and PI staining flow cytometry results showed that LL-37 above 100 μg/mL could significantly reduce the proliferation activity of SV-HUC-1 and increase the proportion of SV-HUC-1 in the G0/G1 phase (P < 0.05), suggesting that cell proliferation was significantly inhibited. Further flow cytometry analysis showed that the calcium ion concentration in SV-HUC-1treated with LL-37 was significantly increased (P < 0.05), and the increase was related to LL-37 concentration. RT-qPCR and WB assays confirmed significant up-regulation of heparan sulfate, a key GAGs component, in the LL-37-treated SV-HUC-1 (P < 0.05).   Conclusion   The cathelicidin LL-37 can inhibit the expression of GAGs and destroy the cell proliferation and transmembrane barrier function of SV-HUC-1.
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