ABSTRACT
Objective:To investigate the role of adenosine diphosphate ribosylation factor 6 (Arf6) in the pathogenesis of endometriosis.Methods:Endometrial tissues were sampled from women who were hospitalized in the Affiliated Hospital of Medical School of Ningbo University and Ningbo Women and Children′s Hospital from November 2020 to May 2021 with endometriosis ( n=44, endometriosis group) and without endometriosis ( n=17, control group). The expression of Arf6 protein in the endometrial tissues was detected by western blot. Endometrial epithelial cells from both groups were primary cultured and the distribution of intracellular mitochondria was detected by immunofluorescence. The expression of Arf6 protein was down-regulated by small interference RNA (siRNA), the distribution of mitochondria in cells with decreased Arf6 protein expression was observed, and the expression of mitochondria-related proteins development and differentiation enhancing factor 1 (DDEF1, also called AMAP1), reactive oxygen species 1 (ROS1) and epithelial-mesenchymal transition (EMT)-related proteins E-cadherin, vimentin were detected. Transwell assay was used to detect the changes in the migration ability of the cells. Results:Compared with the control group, ectopic endometrial tissue of endometriosis group showed high expression of Arf6 protein (0.174±0.019 vs 0.423±0.033; t=29.630, P<0.01); and in ectopic endometrial epithelial cells, mitochondria were distributed near the edge of the cell membrane. While Arf6 expression was down-regulated by siRNA, the distribution of mitochondria in ectopic cells returned to natural, close to the control level. In addition, the expression levels of AMAP1 and ROS1 in ectopic cells after Arf6 protein knockdown were significantly decreased. Transwell assay results indicated that knockdown of Arf6 could reduce the migration ability of ectopic epithelial cells [migration cell count: (34.3±7.5) cells]; and immunofluorescence verified low expression of E-cadherin but high expression of vimentin in ectopic epithelial cells, whereas knockdown of Arf6 protein E-cadherin expression increased but vimentin expression decreased. Conclusions:High expression of Arf6 protein in ectopic endometrial epithelial cells leads to the distribution of mitochondria tending to membrane marginalization, while inducing EMT, which are involved in the mechanism of endoheterosis pathogenesis.
ABSTRACT
Objective To investigate the protective effect of ADP-ribosylation factor 6 inhibitor on acute kidney injury induced by sepsis in mice.Methods In February 2018,thirty male BALB/c mice were divided into uninfected group (5 mice),fluconazole group (5 mice),ADP-ribosylation factor 6 inhibitor group (10 mice)(inhibitor group) and saline control group (10 mice)(control group) by random number table method.In fluconazole group,inhibitor group and control group,1 × 105 CFU of Candida albicans was injected via tail vein for modeling.The uninfected group was injected with equal volume of saline.After 3 hours,inhibitor group was injected with 1.032 mg ADP-ribosylation factor 6 inhibitor,and fluconazole group was injected with 51 μg fluconazole.The control group were injected with equal volume of saline as inhibitor group.After 24hours,serum creatinine,urea nitrogen were measured by kit method.The mice were clinically scored for sepsis severity according to signs and symptoms after treatment and histopathological changing of kidney tissue were observed and scored according to the damage area of renal cortical with hematoxylin-eosin staining.Results The clinical scores,serum creatinine,urea nitrogen and pathological scores of uninfected group were 0,(0.98 ± 0.38) μmol/L,(9.77 ± 0.36) mmol/L,(0.88 ± 0.30),respectively.The fluconazole group were (0.80 ± 0.84),(1.09 ± 0.51) μmol/L,(9.64 ± 0.17) mmol/L,(1.22 ± 0.270),respectively.The inhibitor group were (2.80 ± 1.32),(1.43 ± 0.50) μmol/L,(12.05 ± 1.20) mmol/L,(2.04 ± 0.55),respectively).The control group were (5.20 ± 1.87),(2.96 ± 1.55) μmol/L,(13.94 ± 1.94) mmoL/L,(2.67±0.55).The difference was statistically significant between inhibitor group and the control group both (P ＜ 0.05).Conclusions ADP-ribosylation factor 6 inhibitor reduce acute kidney injury induced by sepsis in mice.
ABSTRACT
As glucose is known to induce insulin secretion in pancreatic beta cells, this study investigated the role of a phospholipase D (PLD)-related signaling pathway in insulin secretion caused by high glucose in the pancreatic beta-cell line MIN6N8. It was found that the PLD activity and PLD1 expression were both increased by high glucose (33.3 mM) treatment. The dominant negative PLD1 inhibited glucose-induced Beta2 expression, and glucose-induced insulin secretion was blocked by treatment with 1-butanol or PLD1-siRNA. These results suggest that high glucose increased insulin secretion through a PLD1-related pathway. High glucose induced the binding of Arf6 to PLD1. Pretreatment with brefeldin A (BFA), an Arf inhibitor, decreased the PLD activity as well as the insulin secretion. Furthermore, BFA blocked the glucose-induced mTOR and p70S6K activation, while mTOR inhibition with rapamycin attenuated the glucose induced Beta2 expression and insulin secretion. Thus, when taken together, PLD1 would appear to be an important regulator of glucose-induced insulin secretion through an Arf6/PLD1/mTOR/p70S6K/Beta2 pathway in MIN6N8 cells.