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OBJECTIVE To study the mechanism of Qingre huashi decoction in the treatment of gastric cancer by intervening in miRNA-155 and inhibiting Wnt/β-catenin signaling pathway. METHODS Thirty nude mice were randomly divided into model group， control group （0.004 g/kg cisplatin+0.02 g/kg fluorouracil）， overexpression group， Qingre huashi prescription low-dose， medium-dose and high-dose groups （2.71， 5.43， 10.86 g/kg）， with 5 mice in each group. The overexpression group was inoculated with miRNA-155 AGS cell line， and the other groups were inoculated with AGS cells to induce tumor-bearing gastric cancer model. The control group was given relevant medicine intraperitoneally， and other groups were given relevant medicine or normal saline intragastrically， once a day， for 3 consecutive weeks. The weight of tumor tissue in nude mice was determined； the pathological morphology of tumor tissue was observed； the miRNA-155 expression， mRNA and protein expressions of Wnt7， β-catenin and T- cell factor-4（TCF-4） in tumor tissue were detected. RESULTS Compared with the model group， the tumor weights of nude mice in the control group， the overexpression group and Qingre huashi decoction high-dose group were significantly reduced （P＜0.05）； mRNA and protein expressions of Wnt7， β -catenin and TCF-4 were significantly decreased （P＜0.05）， while miRNA-155 expression was increased significantly （P＜0.05）. Tumor cells exhibited varying degrees of loose arrangement， shallow nuclear staining， and necrotic foci. CONCLUSIONS Qingre huashi decoction can inhibit the protein and mRNA expressions of Wnt7， β-catenin and TCF-4 in Wnt/β-catenin signaling pathway by up-regulating miRNA-155， thus inhibiting the tumor growth of tumor-bearing nude mice.
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Objective:To investigate the changes of serum micro ribonucleic acid-155 (miR-155) and suppressor of cytokine signaling-1 (SOCS-1) levels in patients with IgA nephropathy and their relationship with renal interstitial fibrosis.Methods:A total of 365 patients with primary IgA nephropathy admitted to Jining First People′s Hospital from January 2009 to June 2018 were selected as the research objects. According to the degree of renal interstitial fibrosis, the patients were divided into T0 group (139 cases), T1 group (124 cases) and T2 group (102 cases). In addition, 361 healthy subjects who had physical examination in our hospital in the same period were selected as the healthy control group. Real-time quantitative polymerase chain reaction (qRT-PCR) was used to detect the expression level of miR-155 in serum of all the subjects; the levels of serum SOCS-1, transforming growth factor-β1 (TGF-β1) and monocyte chemoattractant protein-1 (MCP-1) were detected by enzyme-linked immunosorbent assay (ELISA). Logistic regression model was used to analyze the influencing factors of renal interstitial fibrosis in patients with IgA nephropathy; Pearson method was used to analyze the correlation between serum miR-155, SOCS-1 levels and influencing factors of renal interstitial fibrosis, TGF-β1 and MCP-1; receiver operating characteristic (ROC) curve was used to analyze the predictive value of serum miR-155 and SOCS-1 levels in patients with IgA nephropathy.Results:The levels of systolic blood pressure (SBP), diastolic blood pressure (DBP), 24 h urine protein, serum creatinine, serum uric acid, miR-155, TGF-β1, MCP-1, urinary retinol binding protein (RBP) and acetyl β D-glucosaminidase (NAG) in healthy control group, T0 group, T1 group and T2 group were significantly increased in turn, while the levels of hemoglobin, estimated glomerular filtration rate (eGFR), urinary osmotic pressure and serum SOCS-1 were significantly decreased in turn (all P<0.05). High SBP, high DBP, low hemoglobin, high serum creatinine, high uric acid, high 24-hour urine protein and low eGFR level were independent risk factors of renal interstitial fibrosis in patients with IgA nephropathy (all P<0.05). The serum miR-155 level was positively correlated with TGF-β1, MCP-1, SBP, DBP, serum creatinine, serum uric acid levels and 24 h urine protein, but negatively correlated with SOCS-1, hemoglobin and eGFR levels (all P<0.05). The serum SOCS-1 level was negatively correlated with TGF-β1, MCP-1, SBP, DBP, serum creatinine, uric acid levels and 24 h urine protein, but positively correlated with hemoglobin and eGFR levels (all P<0.05). The area under the curve of predicting the occurrence of renal interstitial fibrosis in patients with IgA nephropathy by serum miR-155 and SOCS-1 combined detection was 0.882, which was significantly larger than that by serum miR-155 and SOCS-1 alone ( P<0.05). Conclusions:The expression of miR-155 is up-regulated and SOCS-1 is down-regulated in IgA nephropathy patients, they may be used as predictors to evaluate the occurrence of renal interstitial fibrosis.
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Objective To investigate microRNAs differential expression and polarization of human macrophages in Toxoplasma gondii infection. Methods The microRNAs differential expression of human macrophages in T. gondii infection was analyzed by microarray, and further validated by qRT-PCR. pEGFP-miR-155 was transfected into THP-1 cells by Lipofectamine M2000 and the transfection ratio was detected by flow cytometry. Flow cytometry was used to detect CD86 molecular on the macrophages. qRT-PCR was used to detect iNOS and IL12 mRNA expression. NO and IL12 expression were then evaluated by using ELISA. Results The miR-155 up-regulated more than 4-fold in T. gondii infected macrophages and enhanced as well as post-infection prolong. pEGFP-miR-155 transfection ratio was 82.6%.compared to cells cultured with T. gondii, pEGFP-miR-155 and miR-155-inhibitor, T. gondii and pEGFP-miR-155 inducement enhanced expression of CD86. Additionally, iNOS and IL12 mRNA were enhanced by qRT-PCR (P<0.05). NO and IL12 expression were increased by ELISA. Conclusion T. gondii infection up-regulates the host miR-155 expression to modulate macrophages polarization to M1.
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Aim To explore the effect of Trillium tschonoskii Maxim (TTM) on the expression of miR-NA-155-3p in rats with brain aging induced by D-gal. Methods Fifty SD rats were divided into five groups randomly. The rats were administered with 0. 9% normal saline (NS) subeutaneously every day in control group, 200 mg • kg • d-1 of D-galactose (D-gal) daily inD-galmodelgroup,and50,100and200mg • kg • d-1 of TTM by gavage 2 hours before D-gal injection everyday in TTM treatment groups for 6 weeks. After 5 weeks, Morris water maze was used to test the ability of spatial learning and memory every day. At the 6th week, rats were sacrificed arid hippocampi were tested by Nissl staining and 8-OHdG immunohistochemical (8-OHdC) staining. The expression of miR-155-3p was determined by Real-time PCR, and the levels of Rheb. mTOR, p-inTOR and p70S6K were detected by Western blot. Results The length of escape latency time and path distance in five groups showed a trend of shortening gradually in the orientation navigation experiments. The average escape latency and the distance in D-gal group were longer than those in control group and TTM group (P <0. 01 and 0. 05) , and the number of crossing platform limes less too (P < 0. 05). The arrangement of neurons was irregular and the intercellular space widened in D-gal group compared with those in TTM group by HE staining. There were more Nissl particles in neurons of the hippocampal CA1 area in con-trol group than that in D-gal group, and TTM treatment could increase the number of Nissl bodies-induced by D-gal. Compared with control group, the fluorescence density of 8-OHdG in D-gal group significantly in-creased (P <0. 01) , while that in TTM group was lower than that in D-gal group (P < 0. 05). The expression of miR-155-3p in the hippocampi in D-gal group was significantly higher than that in normal group (P < 0. 05) , while TTM treatment could alleviate D-gal-in-duced increase of miR-155-3p (P < 0. 05) , followed by an increase of the levels of Rheb and p70S6K, and decrease of mTOR. Conclusions The expression of miR-155-3p increased in the hippocampi of aging rats induced by D-gal. TTM could execute the anti-aging process of brain and down-regulate the level of iniR-155-3p through Rheb/mTOR/p70S6K signaling pathway.
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Objective To investigate microRNAs differential expression and polarization of human macrophages in Toxoplasma gondii infection. Methods The microRNAs differential expression of human macrophages in T. gondii infection was analyzed by microarray, and further validated by qRT-PCR. pEGFP-miR-155 was transfected into THP-1 cells by Lipofectamine M2000 and the transfection ratio was detected by flow cytometry. Flow cytometry was used to detect CD86 molecular on the macrophages. qRT-PCR was used to detect iNOS and IL12 mRNA expression. NO and IL12 expression were then evaluated by using ELISA. Results The miR-155 up-regulated more than 4-fold in T. gondii infected macrophages and enhanced as well as post-infection prolong. pEGFP-miR-155 transfection ratio was 82.6%.compared to cells cultured with T. gondii, pEGFP-miR-155 and miR-155-inhibitor, T. gondii and pEGFP-miR-155 inducement enhanced expression of CD86. Additionally, iNOS and IL12 mRNA were enhanced by qRT-PCR (P<0.05). NO and IL12 expression were increased by ELISA. Conclusion T. gondii infection up-regulates the host miR-155 expression to modulate macrophages polarization to M1.
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Objective To investigate the clinical application of serum miRNA-126,miRNA-155 detection in evaluation of plaque property in the carotid atherosclerotic(CAS)desease.Methods A total of 75 patients with the CAS from May 2015 to May 2017 in the Xianyang Central Hospital and Shiquan Country Chinese Traditional Medicine was chosen,consisted of 35 cases of vulvernable plaque group and 40 cases of stable plaque group.Meanwhile,39 cases of healthy physical examines at the same time were regarded as the control group.The expression levels of serum miRNA-126,miRNA-155 in the groups were detected using the real-time reverse transcription-polymerase chain reaction technique.The largest carotid artery plaque thickness(MAPT)and intima-media thickness(IMT)in the groups were measured using the cervical enhancement CT.Re-sults The results of MAPT and IMT were(3.27±1.01 mm,1.93±0.51 mm)in the vulvernable plaque group and(2.50 ±0.79 mm,1.60±0.26 mm)in the stable plaque group.The carotid artery largest plaque thickness and intima-media thick-ness was higher in the vulvernable plaque group than in the stable plaque group(t=9.76,7.86,P<0.01),and there were significant differenes between the two groups.The expression levels of serum miRNA-126and miRNA-155 were(0.22 ± 0.06,0.87±0.18)in the vulvernable plaque group,(0.50±0.12,0.47±0.10)in the stable plaque group and(0.90±0.19, 0.19±0.05)in the control group.MiRNA-155 expression levels significantly increased in stable plaque group and vulvern-able plaque group compared with in the control group,which increased in the vulvernable plaque group compared with in the stable plaque group,and miRNA-126 expression levels markedly decreased,the differences were statistically significant(F=119.3,102.9,P<0.01).In the vulvernable plaque group,miRNA-126 expression negatively correlated with miRNA-155(r=0.912,P<0.01).miRNA-126 expression levels were inversely associated with the carotid artery largest plaque thickness and the intima-media thickness(r=-0.913,-0.893,P<0.01).While miRNA-155 expression levels were positively corre-lated with them(r=0.899,0.907,P<0.01).Conclusion Serum miRNA-155,miRNA-126 detection can be applied to pre-diction of CAS plaques rupture,and may become a useful warning marker of ischemic stroke events.
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PURPOSE: Insufficient sensitivity and specificity prevent the use of most existing biomarkers for early detection of breast cancer. Recently, it was reported that serum microRNAs (miRNAs) may be potential biomarkers in many cancer diseases. In this study, we investigated whether serum levels of 5 miRNAs including miR-21, miR-125b, miR-145, miR-155, and miR-365 could discriminate breast cancer patients and healthy controls. METHODS: Serum levels of miRNAs were measured by using quantitative real-time polymerase chain reaction in 99 breast cancer patients and 21 healthy controls. The abundance change of serum miRNAs were also evaluated following surgical resection in 20 breast cancer patients. Receiver operating characteristic (ROC) curve analysis was performed to assess the sensitivity and specificity of miRNAs as diagnostic biomarkers. RESULTS: Serum levels of miR-21 and miR-155 was significantly higher, while miR-365 was significantly lower in breast cancer as compared with healthy controls. The serum levels of miR-21 and miR-155 significantly decreased following surgical resection. Additionally, the serum level of miR-155 at stages I and II was significantly higher compared to stage III. The serum miR-145 level was remarkably higher in progesterone receptor (PR)-positive patients than PR-negative. The positivity of miR-21, miR-155, and miR-365 was high compared to CA 153 and CEA in breast cancer. ROC curve analyses of a combination of miR-21, miR-155, and miR-365 yielded much higher area under curve and enhanced sensitivity and specificity in comparison to each miRNA alone. CONCLUSION: The combination of serum miR-21/miR-155/miR-365 may potentially serve as a sensitive and specific biomarker that enables differentiation of breast cancer from healthy controls.