The regulation mechanism of phloroglucinol in urinary smooth muscle spasm / 中国药理学通报

Aim To study the regulation and mechanism of phloroglucinol in bladder smooth muscle spasm. Methods In vitro the experiment used bladder muscle strip to verify the relieving effect of phloro-glucinol on bladder spasm by different drugs. At the same time,RT-qPCR and Western blot were used to detect the expression levels of genes involved in the calcium signaling pathway caused by the antispasmodic effect of phloroglucinol. Results Phloroglucinol could relieve bladder spasm, and the antispasmodic effect was enhanced with the increase of concentration, and the expression of calponin 1 and MYLK3 in tissue cells increased. The results of RT-qPCR showed that the expression of Gprc5b G,Ppp2r5a, Chptl, Prkar2b ,Abcd2 and Rasdl genes in mouse bladder tissue significantly decreased, which was consistent with the sequencing results of RNA-seq.Conclusions Phloroglucinol can relieve bladder smooth muscle spasm, and its mechanism is related to calcium signaling pathway. Meanwhile, phloroglucinol also inhibits the expression of Rasdl gene, suggesting that it may be related to cell cycle , protein phosphorylation, choline metabolism, ATP synthesis and tumor-related pathways.

Inhibition of LIM kinase reduces contraction and proliferation in bladder smooth muscle

Overactive bladder (OAB) is the most bothersome symptom in lower urinary tract symptoms (LUTS). Current pharmacologic treatment aims to inhibit detrusor contraction; however, shows unsatisfied efficacy and high discontinuation rate. LIM kinases (LIMKs) promote smooth muscle contraction in the prostate; however, their function in the bladder smooth muscle remains unclear. Here, we studied effects of the LIMK inhibitors on bladder smooth muscle contraction and proliferation both

Effect of acetylation modification on microenvironment adaptability of mesenchymal stem cells / 解放军医学杂志

Objective To study the changes of acetylation level of smooth muscle marker gene after the differentiation of bone marrow mesenchymal stem cells (BMSCs) in smooth muscle microenvironment, and explore the role and mechanism of histone acetylation modification in the differentiation of stem cells. Methods BMSCs and bladder smooth muscle cells (BSMCs) were cultured in vitro. The third generation BMSCs of the same batch were selected, and BMSCs co-cultured with BSMCs for 3 days were set as the experimental group and the BMSCs without co-culture as the control group. RT-PCR was performed to compare the expression abundance between the two groups of α-smooth muscle actin (α-SMA), calponin and smooth muscle myosin heavy chain (SM-MHC) in BMSCs. Ultrasound of power 80%, 20 times, 0.5 s and 8 cycles were used to break the BMSCs DNA of both experimental and control group. Antibody H3K9 was used to bind to the specific acetylation sites. The acetylation site genes of histone in BMSCs were precipitated by chromatin immunoprecipitation (ChIP). The genes obtained was amplified by adaptor PCR. The expression levels of the three kinds of target genes (α-SMA, calponin, SM-MHC) were detected by Real-time PCR. Results RT-PCR showed that the expression levels of mRNA of smooth muscle marker genes (α-SMA, calponin, SM-MHC) in BMSCs were significantly higher in experimental group than in control group (0.176±0.003 vs. 0.070±0.002; 0.079±0.002 vs. 0.051±0.003 and 0.091±0.004 vs. 0.034±0.001, respectively) with significant differences. Test results of spectrophotometer showed that the amount of DNA obtained by precipitation of H3K9 acetylated antibody was higher than that of IgG antibody, and was higher in experimental group than in control group (P<0.05). Real-time PCR used to analyze the acetylation level of H3K9 before and after the differentiation of BMSCs showed that the mRNA transcription levels of smooth muscle marker genes (α-SMA, calponin, SM-MHC) were significantly higher in experimental group than in the control group (9.26±5.03 vs. 1.01±0.05, 2.33±0.65 vs. 0.99±0.05, 2.63±0.37 vs. 1.00±0.03, respectively) after BMSCs differentiation with statistically significant differences. Conclusion The increase of acetylation level of H3K9 specific site of BSMCs in smooth muscle microenvironment promotes the differentiation of BMSCs into BSMCs.

Application of fluorescence recovery after photobleaching in the detection of gap junctional intercellular communication in bladder smooth muscle / 第三军医大学学报

Objective To explore the functional changes of gap junctional intercellular communication(GJIC) in bladder smooth muscle. Methods The functions of GJIC in bladder smooth muscle were detected by fluorescence recovery after photobleaching(FRAP). The mean fluorescence recovery rates of the bladder smooth muscle cells in the experimental group and the control group were compared. Results The mean fluorescence recovery rate of the experimental group was significantly higher than that in the control group( P

Biocompatibility of vessel extracellular matrix with bladder smooth muscle cells of rabbits / 中华泌尿外科杂志

Objective To evaluate the biocompatibility of vessel extracellular matrix (VECM) with bladder smooth muscle cells of rabbits,and to discuss the feasibility of vessel extracellular matrix as a matrix for urinary tract reconstruction.Methods Primary cuhured bladder smooth muscle cells (RBSMCs) iso- lated from New Zealand rabbits were implanted on VECM (1?10~6 cells/ml).The effect of VECM on meta- bolic activity,attachment,proliferation of RBSMCs were monitored in vitro by inverted light microscopy and scanning electron microscopy.The extracts of VECM and emulsion were prepared as experimental group and positive controls separately.The culture medium was used as negative control,and simple culture medium without cells was used as blank control.The cell viability was monitored by MTT method after 1-,3-,5-d see- ding.The in vivo tissue response to VECM was investigated by implanting into the subcutaneous sites of the rabbits.Results VECM exhibited nontoxic and bioactive effect on RBSMCs.RBSMCs could be attached to and proliferated on VECM and remained their morphologies.The cell proliferation rates of experimental group were 95.61%、98.34%、102.91%,respectively,after 1,3,5 d;those of negative control group were 100.00% ,respectively;and those of positive control group were 35.14%、38.95%、32.66%,respectively. There was significant difference in the rate between experimental group and positive control (P＜0.01),and no significant difference in the rate between experimental group and negative control (P＞0.05).In vivo, VECM demonstrated favorable tissue compatibility without tissue necrosis and fibrosis.Conclusions VECM exhibits nontoxic and bioactive effects on primary cultured bladder smooth muscle cells.It is a suit- able material for urinary tract reconstruction.