| Objective: Pelvic organ prolapse(POP)is a kind of disease caused by the defect or injury of pelvic floor supporting structure.The main pathological reason is the weakness of connective tissue in the supporting structure.The principal manifestation of the disease is pelvic organs prolapse from the pelvic cavity.It rarely affects the quality of women’s life.POP occurs usually in older women,it has a significant impact on the quality of life in those women although rarely causing death.Its pathogenesis has not been clearly explained.The supporting function of the pelvic floor system is mainly due to collagen in connective tissue.If collagen content is reduced and structure changes,POP can occur.Increasing of apoptosis of fibroblast can lead to reduced of collagen content in connective tissue.Transforming growth factor-β1(TGF-β1)is an important fibrosis promoter,which is mainly expressed in fibroblasts and can promote the secretion of collagen.TGF-β1 works through the TGF-β1/Smad signaling pathway.Smad is a downstream factor of TGF-β1,which precisely regulates cell homeostasis and transcriptional responses through phosphorylation of molecules in specific regions.Casein kinase-2 interacting protein-1(CKIP-1),is also known as PLEKHO1(Pleckstrin homology protein family O subfamily member1).CKIP-1 can inhibit the activity of TGF-β1 and participate in the regulation of apoptosis.In POP,it remains to be clarified that whether CKIP-1 can affect collagen secretion of fibroblasts through TGF-β1 /Smad3 signaling pathway or can regulate apoptosis of fibroblasts.We predict that mi R-98-5p and CKIP-1 have binding sites through bioinformatics software http://www.targetscan.org/ and the literature.We also predict that mi R-98-5p and Lnc RNA NEAT1 have binding sites through bioinformatics database https://starbase.sysu.edu.cn and the literature.Lnc RNAs often competitively bind with micro RNAs to regulate downstream proteins.It remains to be further clarified that whether Lnc RNA NEAT1 regulate the expression of CKIP-1 through binding with mi R-98-5p competitively to influence secretion of collagen and apoptosis of fibroblasts.Methods: 1.30 cases of vaginal wall tissue of POP group and control group were collected respectively.HE staining and Masson staining were used to detect histopathological change and collagen deposition of the vaginal wall of the two groups.TUNEL was used to detect apoptosis between the two groups.Western blot was used to detect the expressions of apoptosis markers Bax and Bcl-2.Immunohistochemistry and Western blot were used to detect the expressions of CKIP-1,TGF-β1,Smad3,p-Smad3,COL-I and COL-III.Cell transfection technology was used to overexpress and knockdown of CKIP-1,the expressions of TGF-β1,Smad3,p-Smad3,COL-I,COL-III and apoptosis markers Bax and Bcl-2 were detected by Western blot.2.RT-PCR was used to detect the expression of mi R-98-5p in vaginal wall tissue of POP group and control group,and to analyze the correlation between mi R-98-5p and CKIP-1.Cell transfection technology was used to overexpress and knockdown of mi R-98-5p,the expressions of CKIP-1,TGF-β1,Smad3,p-Smad3,COL-I,COL-III,Bax and Bcl-2 were detected by Western blot.Vaginal wall fibroblasts were transfected with mi R-98-5p mimics and Ov-CKIP-1,the expressions of CKIP-1,TGF-β1,Smad3,p-Smad3,COL-I,COL-III,Bax and Bcl-2 were detected by Western blot.3.RT-PCR was used to detect the expression of Lnc RNA NEAT1 in vaginal wall tissue of POP group and control group,and to analyze the correlation between Lnc RNA NEAT1 and mi R-98-5p.Cell transfection technology was used to overexpress and knockdown of Lnc RNA NEAT1,the expressions of CKIP-1,TGF-β1,Smad3,p-Smad3,COL-I,COL-III,Bax and Bcl-2 were detected by Western blot.Vaginal wall fibroblasts were transfected with NEAT1 inhibitor and mi R-98-5p inhibitor,the expressions of CKIP-1,TGF-β1,Smad3,p-Smad3,COL-I,COL-III,Bax and Bcl-2 were detected by Western blot.Results: 1.The collagen fibers in the POP group were broken,discontinuous,disordered and loose,while the collagen fibers in the control group were tight and orderly.Collagen fiber content was less in the POP group.The levels of apoptosis in the vaginal wall of the POP group are increased.The expressions of apoptosis markers Bax was higher and Bcl-2 was lower in the POP group.The expression of CKIP-1 in vaginal wall of the POP group was higher than that in the control group,while the expressions of TGF-β1,Smad3,p-Smad3,COL-I and COL-III were lower than that in the control group.After overexpression of CKIP-1,the expressions of TGF-β1,Smad3,p-Smad3,COL-I,COL-III and Bcl-2 decreased,while the expression of Bax increased.After knockdown of CKIP-1,the expressions of TGF-β1,Smad3,p-Smad3,COL-I,COL-III and Bcl-2increased,while the expression of Bax decreased.2.The expression of mi R-98-5p in vaginal wall of the POP group was lower,mi R-98-5p was negatively correlated with CKIP-1 expression.After overexpression of mi R-98-5p,the expressions of TGF-β1,Smad3,p-Smad3,COL-I,COL-III and Bcl-2increased,while the expressions of CKIP-1 and Bax decreased.After knockdown of mi R-98-5p,the expressions of TGF-β1,Smad3,p-Smad3,COL-I,COL-III and Bcl-2decreased,while the expressions of CKIP-1 and Bax increased.After fibroblasts were transfected with mi R-98-5p mimics and ov-CKIP-1,the expressions of TGF-β1,Smad3,p-Smad3,COL-I,COL-III and Bcl-2 decreased,while the expressions of CKIP-1 and Bax increased.3.The expression of Lnc RNA NEAT1 in vaginal wall of the POP group was higher,Lnc RNA NEAT1 was negatively correlated with mi R-98-5p.After overexpression of NEAT1,the expressions of TGF-β1,Smad3,p-Smad3,COL-I,COL-III and Bcl-2decreased,while the expressions of CKIP-1 and Bax increased.After knockdown of NEAT1,the expressions of TGF-β1,Smad3,p-Smad3,COL-I,COL-III and Bcl-2increased,while the expressions of CKIP-1 and Bax decreased.After fibroblasts were transfected with NEAT1 inhibitor and mi R-98-5p inhibitor,the expressions of TGF-β1,Smad3,p-Smad3,COL-I,COL-III and Bcl-2 decreased,while the expressions of CKIP-1and Bax increased.Conclusions: 1.The expressions of COL-I and COL-III in the vaginal wall of pelvic organ prolapse were decreased.The levels of apoptosis in the vaginal wall of the POP group are increased.CKIP-1 can decrease the expressions of COL-I and COL-III through TGF-β1/Smad3 signaling pathway,and can promote apoptosis of fibroblasts.2.The expression of mi R-98-5p in the vaginal wall of pelvic organ prolapse was decreased.The low expression of mi R-98-5p can increase the expression of CKIP-1,then CKIP-1 can decrease the expressions of COL-I and COL-III through TGF-β1/Smad3 signaling pathway,and can promote apoptosis of fibroblasts.3.The expression of Lnc RNA NEAT1 was increased in the vaginal wall of pelvic organ prolapse,NEAT1 can up-regulate the expression of CKIP-1 by inhibiting mi R-98-5p,and then CKIP-1 can decrease the expressions of COL-I and COL-III through TGF-β1/Smad3 signaling pathway,and can promote apoptosis of fibroblasts. |
PDF Full Text Request