Inducible nitric oxide synthase (iNOS) is a class of catalytic enzyme molecules that play a regulatory role in the development of many diseases including bladder fibrosis. To investigate the effects of cytokines interleukin-1 α (IL-1 α), interleukin-1 β (IL-1 β), interferon- γ (IFN- γ) on bladder fibrosis induced by iNOS in human bladder smooth muscle cells （hb-SMCs）. Evaluate the effect of inflammation on the deterioration of neurogenic bladder function.

To remove hb-SMCs for cell resuscitation or subcultivation and continue incubating in 5% CO2 and 37°C incubator. Change medium every 2-3 days. Take the 5th to 7th generation cells and wait for the cells to grow about 80% to 90% at the bottom of the dish or when the cells are growing, replace the old medium with 1% fetal bovine serum in DMEM medium and starve the cells for 24 hours. They were randomly divided into 1 experimental group: 100 ng/ml IFN-γ, 20 ng/ml IL-1α and 20 ng/ml IL-1β mixture + DMEM medium were added to the culture; 2 blank control groups: hb-SMCs were not treated Only DMEM complete medium was added. The expression of NF-κB p65, iNOS, and Collagen III protein in hb-SMCs was detected by cell fluorescence immunofluorescence and Western Blot after 2 days in vitro. Q-PCR was used to detect NF-κB p65, iNOS in hb-SMCs. And Collagen III mRNA expression.

Analysis of cell fluorescence, Western Blot, and Q-PCR assays showed that the expression of NF-κB p65, INOS, and Collagen III protein and mRNA in the hb-SMCs of the experimental group was significantly higher than that of the control group (p<0.05).

The effect of cytokine mixture on the expression of iNOS、NF-κB p65 and Collagen III in hb-SMCs may be one of the mechanisms of urinary inflammation in the development of bladder fibrosis. If the bladder of a patient with neurogenic bladder is chronically infiltrated with cytokines, the bladder function may deteriorate.

Bladder fibrosis is one of the common clinical problems in urology. It can cause changes in bladder function and structure and ultimately lead to damage to the upper urinary tract. There is currently no effective treatment for the treatment or reversal of fibrosis. The study found that bladder fibrosis is related to bladder inflammation, bladder pressure and bladder blood supply. In this study, we designed a mixture of inflammatory cytokines to stimulate hb-SMCs, and to observe the expression changes of NF-κB, iNOS, and Collagen III to explore inflammation. Possible mechanisms of action in bladder fibrosis. The following are preliminary conclusions: The inflammatory cytokines IL-1α, IL-1β and IFN-γ may initiate collagen-III expression through activation of NF-κB, induce iNOS transcription, promote bladder fibrosis, which may be one of the mechanisms of its development. . Patients with neurogenic bladder are prone to urinary tract infection. Long-term infiltration of cytokines may be one of the causes of worsening bladder function and worsening symptoms in NB patients.