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AIM To investigate the effect of sinomenine (SN) on the damage of human neuroblastoma SK-N-SH cells induced by 1-methyl-4-4 phenylpyridine (MPP+) and its mechanism for exploring the pathogenesis of Parkinson disease. METHODS SN was used to treat MPP+-induced SK-N-SH cells. The levels of malondialdehyde (MDA) and glutathione (GSH) in cell culture supernatants were measured by ELISA. The apoptosis was analyzed by flow cytometry. The protein expression levels of Bcl-2 and Bax were determined by Western blot. The expression levels of long noncoding RNA ANRIL and microRNA-626 (miR-626) were detected by RT-qPCR. Dual-luciferase reporter assay was used to evaluate the relationship between ANRIL and miR-626. After ANRIL small interfering RNA was transfected into SK-N-SH cells, the effects of ANRIL expression knock-down on MPP+-induced SK-N-SH cell apoptosis, the protein expression levels of Bcl-2 and Bax, and the levels of MDA and GSH in cell culture supernatants were examined. RESULTS After treatment with MPP+, the apoptotic rate, Bax protein level and ANRIL expression in SK-N-SH cells were increased (P <0.05), and the Bcl-2 protein level and miR-626 expression were decreased (P <0.05). The level of MDA in cell culture supernatants was increased (P <0.05), and the level of GSH was decreased (P <0.05). After SN treatment or ANRIL expression knock-down, decreased apoptotic rate, Bax protein level and ANRIL expression (P <0.05), and increased Bcl-2 protein level and miR-626 expression in MPP+-induced SK-N-SH cells were observed (P <0.05). The level of MDA in the cell culture supernatants was decreased (P <0.05), and the level of GSH was increased (P <0.05). CONCLUSION SN attenuates MPP+-induced damage in SK-N-SH cells by regulating ANRIL/miR-626 signaling pathway. 相似文献
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AIM: To investigate the effect of microRNA-323 (miR-323) on the apoptosis of hypoxia-induced rat H9C2 cardiomyocytes and its mechanism. METHODS: The hypoxic injury model was established in the H9C2 cells. Anti-miR-323, pcDNA-FGF9 and si-FGF9 were transfected into the H9C2 cells and cultured under hypoxic condition for 48 h. The expression of miR-323 was detected by qPCR. The protein levels of fibroblast growth factor 9 (FGF9), cleaved caspase-3, c-Jun N-terminal kinase (JNK) and p-JNK were determined by Western blot. The cell viability was measured by MTT assay, and the apoptosis was analyzed by flow cytometry. The method of bioinformatics was applied to predict the target gene of miR-323, and dual-luciferase reporter assay was used for further validation. RESULTS: Hypoxia greatly reduced the viability of H9C2 cells at 12 h, 24 h and 48 h (P <0.05), and remarkably increased apoptotic rate and the protein level of cleaved caspase-3 in a time-dependent manner (P <0.05). The expression of miR-323 and the protein level of p-JNK were up-regulated and the expression of FGF9 was down-regulated in the H9C2 cells exposed to hypoxia (P <0.05). Down-regulation of miR-323 and over-expression of FGF9 obviously increased the viability of the H9C2 cells exposed to hypoxia, and decreased the apoptotic rate and the protein level of cleaved caspase-3 (P <0.05). FGF9 was the target gene of miR-323. Down-regulation of FGF9 reversed the attenuating effect of down-regulation of miR-323 on hypoxia-induced H9C2 cell injury. miR-323 regulated FGF9 and affected p-JNK level. CONCLUSION: miR-323 affects the viability and apoptosis of H9C2 cardiomyocytes under hypoxia by targeting FGF9 and regulating JNK signaling pathway. 相似文献
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AIMTo determine the effect of microRNA-206 (miR-206) on proliferation and migration of human papillary thyroid carcinoma K1 cells and to explore its possible mechanism. METHODSThe expression of miR-206 in the K1 cells was detected by RT-qPCR. The cell viability was detected by CCK-8 assay. The number of viable K1 cells was counted by the method of Trypan blue exclusion. The migration ability of K1 cells was detected by Transwell chamber migration assay. Bioinformatics software was used to predict the target gene of miR-206. The targeting relationship between miR-206 and c-Met was verified by dual-luciferase reporter assay. The protein levels of c-Met, p-Met, AKT, p-AKT, mTOR and p-mTOR were determined by Western blot. RESULTSAfter the K1 cells were transfected with miR-206 mimic transiently, the relative expression of miR-206 in treatment group was significantly higher than that in blank group and negative control group (P <0.01). The results of CCK-8 assay and Trypan blue exclusion assay showed that the proliferation ability of K1 cells in treatment group transfected with miR-206 mimic was significantly inhibited compared with other groups (P <0.01). The results of Transwell assay showed that the number of migratory K1 cells in treatment group was lower than that in blank group and negative control group (P <0.01). Moreover, our results demonstrated that miR-206 directly targeted c-Met and repressed the activation of downstream AKT/mTOR signaling pathway. CONCLUSION miR-206 over-expression inhibits the proliferation and migration abilities of papillary thyroid carcinoma K1 cells, and its mechanism may be related to the inhibition of c-Met/AKT/mTOR signaling pathway. 相似文献
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AIM To investigate the protective effect of resveratrol (Res) on cortical neurons in rat bacterial meningitis (BM) model. METHODS Group B hemolytic Streptococcus was injected via the posterior cistern to establish a BM model. Resveratrol was administered intranasally and microRNA-223-3p (miR-223-3p) antagomir was administered by intracerebroventricular injection. HE staining was used to observe the pathological changes of the brain tissue. Loeffler scoring method was used to evaluate the neurobehavioral functions. TUNEL staining was used to detect neuronal apoptosis. The expression of interleukin-1β (IL-1β), IL-18, glial fibrillary acidic protein (GFAP) and ionized calcium-binding adaptor molecule 1 (Iba1) was detected by immunofluorescence staining. The protein levels of nucleotide-binding oligomerization domain-like receptor protein 3 （NLRP3）, cleaved caspase-1, IL-1β and IL-18 were determined by Western blot. The expression level of miR-223-3p was detected by RT-qPCR. Online software TargetScan was used to search for the complementary nucleotide sequences between miR-223-3p and NLRP3 mRNA. RESULTS Compared with sham group, the thickness of meninges in BM model was increased, the neurological score was decreased (P <0.05), and the number of TUNEL positive neurons was increased significantly (P <0.05). Astrocytes and microglia were activated, the fluorescence intensity of IL-1β and IL-18 was increased (P <0.05), and the expression levels of NLRP3, cleaved caspase-1, IL-1β, IL-18 and miR-223-3p were increased (P <0.05). Compared with BM group, after treatment with resveratrol, the neurological score was increased (P <0.05), the number of TUNEL positive neurons was decreased significantly (P <0.05), and the inflammatory response of astrocytes and microglia was suppressed. The fluorescence intensity of IL-1β and IL-18 was decreased (P <0.05), the protein levels of NLRP3, cleaved caspase-1, IL-1β and IL-18 were decreased (P <0.05), and the expression level of miR-223-3p was increased (P <0.05). A nucleotide sequence in the 3'-UTR of NLRP3 mRNA might be targeted by miR-223-3p. In the brain of rat BM model, compared with antagomir control group, the expression of NLRP3 was increased in miR-223-3p antagomir group with resveratrol treatment (P <0.05). CONCLUSION Resveratrol may reduce the inflammatory death of cortical neurons in BM model of infant rats through miR-223-3p/NLRP3 pathway, thus playing a protective role for the neurons. 相似文献
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AIM To study the effects of extracts of Herba Taxilli (Sangjisheng, SJS) on the viability and apoptosis of osteoarthritic chondrocytes and the underlying mechanism. METHODS Human primary osteoarticular chondrocytes (RPOC) were divided into control group, interleukin-1β (IL-1β) group, IL-1β+low-dose extracts of SJS (SJS-L) group, IL-1β+medium-dose extracts of SJS (SJS-M) group, IL-1β+high-dose extracts of SJS (SJS-H) group, IL-1β+anti-miR-NC group, IL-1β+anti-miR-375 group, IL-1β+SJS-H+miR-NC group, IL-1β+SJS-H+miR-375 group. The cell viability was measured by MTT assay, apoptosis was analyzed by flow cytometry, miR-375 expression was detected by qPCR, and the protein levels of cyclin D1, P21, Bcl-2, Bax and caspase-3 were determined by Western blot. RESULTS Compared with control group, the viability of RPOC at 24 h, 48 h and 72 h and the protein expression levels of cyclin D1 and Bcl-2 were significantly decreased (P <0.05), the protein levels of P21, Bax and caspase-3, the apoptotic rate and the expression level of miR-375 were remarkably increased in IL-1β group(P <0.05). Compared with IL-1β group, the cell viability at 24 h, 48 h and 72 h and the protein expression of cyclin D1 in the RPOC were greatly increased (P <0.05), while the expression of P21 was significantly decreased in IL-1β+SJS-M group and IL-1β+SJS-H group(P <0.05).The apoptotic rate, Bax, caspase-3 protein and miR-375 expression were obviously decreased (P <0.05), and Bcl-2 protein level was significantly increased in IL-1β+SJS-H group compared with IL-1β group(P <0.05). Compared with IL-1β+anti-miR-NC group, the expression of miR-375, the protein levels of P21, Bax, caspase-3 and the apoptotic rate in the RPOC of IL-1β+anti-miR-375 group were markedly decreased (P <0.05), while the cell viability at 24 h, 48 h and 72 h and the protein levels of cyclin D1 and Bcl-2 were significantly increased (P <0.05). Over-expression of miR-375 reversed the effects of extracts of SJS on the viability and apoptosis of RPOC with IL-1β stimulation. CONCLUSION The extracts of Herba Taxilli promotes the viability and inhibits apoptosis of RPOC treated with IL-1β, which is related to the regulation of miR-375 expression. 相似文献
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AIM: To investigate the effect of microRNA-214-5p (miR-214-5p) on myocardial injury and immune response in rats with ischemia-reperfusion (I/R) by targeting p21-activated protein kinase 4 (PAK4). METHODS: The rats were divided into sham group, I/R group, Ad-Scramble group, and Ad-miR-214 group (n =9). Adenovirus was injected into 6 different sites on the anterior wall of the left ventricle of the rats. Four days later, the I/R model was constructed by suturing the left anterior descending coronary artery. The expression level of miR-214 was detected by RT-qPCR. Myocardial injury was observed by HE staining. The levels of heart damage markers (CK-MB, Mb, and cTnI) and inflammatory factors (IL-6, IL-1β and TNF-α) were measured by ELISA. The rate of cardiomyocyte apoptosis was analyzed by flow cytometry. The content of MDA and the activity of SOD were detected by commercially available kits. Target genes were predicted by genetic software and verified by dual-luciferase reporter assay. The protein levels of caspase-3, caspase-9, Bcl-2, Bax, PAK4, p-Akt and p-mTOR were determined by Western blot. RESULTS: miR-214 was down-regulated in the cardiomyocytes of I/R rats (P <0.01). Over-expression of miR-214-5p attenuated myocardial injury in the I/R rats, down-regulated the expression of CK-MB, Mb and cTnI, decreased the apoptotic rate of cardiomyocytes, up-regulated the expression of Bcl-2, down-regulated Bax, caspase-3 and caspase-9 expression, increased SOD activity, and decreased the content of MDA, IL-6, IL-1β and TNF-α (P <0.01). The binding sites of miR-214-5p and PAK4 were pre-sent in the 3’-UTR, and over-expression of miR-214-5p up-regulated the protein levels of PAK4, p-Akt and p-mTOR (P <0.01). CONCLUSION: miR-214-5p over-expression attenuates myocardial injury in I/R rats by targeting PAK4, inhibits cardiomyocyte apoptosis, oxidative stress and inflammation, and activates the PI3K/Akt/mTOR pathway. 相似文献
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AIM To investigate the mechanism of long noncoding RNA （lncRNA） FEZF1-AS1 regulating microRNA-363-3p （miR-363-3p） on the viability and apoptosis of lipopolysaocharide （LPS）-induced vascular endothelial cells. METHODS Human umbilical vein endothelial cells （HUVECs） were cultured in vitro . pcDNA-NC, pcDNA-FEZF1-AS1, anti-miR-NC, anti-miR-363-3p, miR-NC and miR-363-3p mimics were transfected into the HUVECs and LPS stimulation was applied for 24 h. RT-qPCR was used to detect the expression of FEZF1-AS1 and miR-363-3p. The cell viability was measured by MTT assay. The apoptotic rate was analyzed by flow cytometry. The dual-luciferase reporter experiment was used to verify the targeted regulation of FEZF1-AS1 and miR-363-3p. Western blot was used to determined the expression of cyclin D1, Ki67 and cleaved caspase-3. RESULTS Compared with control group, the expression level of FEZF1-AS1 in LPS group was significantly reduced （P <0.05）, and the expression level of miR-363-3p was significantly increased （P <0.05）. Compared with pcDNA-NC+LPS group, the cell viability in pcDNA-FEZF1-AS1+LPS group was significantly increased （P <0.05）, the apoptotic rate was significantly reduced （P <0.05）, the protein levels of cyclin D1 and Ki67 were significantly increased （P <0.05）, and the protein level of cleaved caspase-3 was significantly reduced （P <0.05）. Compared with anti-miR-NC+LPS group, the cell viability in anti-miR-363-3p+LPS group was significantly increased （P <0.05）, the apoptotic rate was significantly reduced （P <0.05）, the protein levels of cyclin D1 and Ki67 were significantly increased （P <0.05）, and the protein level of cleaved caspase-3 was significantly reduced （P <0.05）. Dual-luciferase reporter experiment confirmed that FEZF1-AS1 targeted miR-363-3p. Compared with miR-NC+pcDNA-FEZF1-AS1+LPS group, the cell viability in miR-363-3p+pcDNA-FEZF1-AS1+LPS group was significantly reduced （P <0.05）, the apoptotic rate was significantly increased （P <0.05）, the protein levels of cyclin D1 and Ki67 were significantly reduced （P <0.05）, and the protein level of cleaved caspase-3 was significantly increased （P <0.05）. CONCLUSION Over-expression of FEZF1-AS1 promotes the viability and inhibits apoptosis of LPS induced vascular endothelial cells by inhibiting the expression of miR-363-3p. 相似文献
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AIM To investigate the effect of NOD-like receptor family caspase recruitment domain containing 3 (NLRC3 ) expression knock-down on the viability and apoptosis of normal human bronchial epithelial BEAS-2B cells and its mechanism. METHODS The small interfering RNA (siRNA) fragments of NLRC3 gene were transfected into BEAS-2B cells using Lipofectamine 2000 transfection reagent to knock down the NLRC3 expression. The interference fragment was screened by RT-qPCR. The cell viability was measured by MTT assay. The mitochondrial membrane potential was detected by JC-1 staining. The apoptotic rate was analyzed by flow cytometry with annexin V-FITC/PI staining. The protein expression levels of Bcl-2 and Bax were determined by Western blot. RESULTS The interference segment 3 of NLRC3 gene (siNLRC3-3) displayed the best interference effect on NLRC3 expression in BEAS-2B cells (P <0.01). Knock-down of NLRC3 expression in BEAS-2B cells enhanced the cell viability (P <0.01). Knock-down of NLRC3 increased the mitochondrial membrane potential, and decreased the apoptotic rate (P <0.05). Moreover, knock-down of NLRC3 significantly up-regulated Bcl-2 protein expression and significantly down-regulated Bax protein expression (P <0.01). CONCLUSION Knock-down of NLRC3 expression enhances the viability and inhibits the apoptosis of BEAS-2B cells, which may be related to increase in the expression of Bcl-2 protein and decrease in the expression of Bax protein. 相似文献
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SUN Xin-feng HAN Zhi-yi FENG Wen-xing MA Wen-feng ZHANG Wei ZHOU Xiao-zhou 《园艺学报》2000,36(8):1487-1492
AIM To investigate the effect of paeonol on the viability and migration ability of hepatocellular carcinoma cells and its molecular mechanism. METHODS Human hepatocellular carcinoma Hep3B cells was treated with paeonol at different concentrations (50, 100, 200 and 400 mg/L). The cell viability was measured by CCK-8 assay to determine the optimal drug concentration. The Hep3B cells were divided into normal control (NC) group, paeonol group, miR-NC group, miR-424-3p group, paeonol+anti-miR-NC and paeonol+anti-miR-424-3p group. The expression level of miR-424-3p was detected by RT-qPCR. The migration ability was detected by Transwell assay. The protein levels of cyclin D1, matrix metalloproteinase 2 (MMP2), MMP9 and PI3K/AKT signaling pathway-related molecules were determined by Western blot. RESULTS Paeonol intervention inhibited the viability of Hep3B cells in a concentration-dependent manner (P <0.05). The concentration of paeonol at 200 mg/L was selected for the following study. Paeonol intervention inhibited the protein expression of MMP2 and MMP9 in the Hep3B cells, and inhibited the migration ability of the Hep3B cells. Paeonol intervention promoted the expression of miR-424-3p in the Hep3B cells (P <0.05). Over-expression of miR-424-3p inhibited the expression of cyclin D1, MMP2 and MMP9 in the Hep3B cells and inhibited cell viability and migration ability (P <0.05). Inhibition of miR-424-3p reversed the effect of paeonol on the viability and migration ability of the Hep3B cells (P <0.05). Paeonol inhibited phosphorylation levels of PI3K and AKT in the Hep3B cells and inhibited the activation of PI3K/AKT signaling pathway (P <0.05). Inhibition of miR-424-3p reversed the effect of paeonol on PI3K/AKT signaling pathway in the Hep3B cells (P <0.05). CONCLUSION Paeonol inhibits the viability and migration ability of hepatocellular carcinoma cells by up-regulating miR-424-3p and inhibiting PI3K/AKT signaling pathway. 相似文献
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AIM To investigate the role of fatty acid translocase (FAT/CD36) on differentiation of monocytes to macrophages. METHODS Human monocyte THP-1 cells were treated with phorbol 12-myristate 13-acetate (PMA) at 0, 100 and 200 μg /L. Small interfering RNA (siRNA) targeting CD36 (siCD36) was employed to knock down the expression of CD36 in THP-1 cells. The CD36 over-expression (CD36OE) cell line was constructed by transfection with a recombinant lentivirus containing CD36 cDNA. Optical microscopy and crystal violet staining were used to detect the monocyte morphological changes and adhesion ability. The protein expression of CD36 was measured by flow cytometry and Western blot. The mRNA levels of CD36, CD11b and CD80 were detected by real-time PCR. The protein levels of extracellular signal-regulated kinase (ERK） and Src tyrosine kinase were determined by Western blot. RESULTS The cellular adhesiveness of THP-1 cells was elevated in the process of monocytes differentiation, and the expression of CD36 was increased in this process as well (P <0.01). siCD36 was transfected into the THP-1 cells (CD36i group) and the silencing efficiency was approximately 80%. The cell surface area and cellular adhesiveness were significantly decreased in CD36i group compared with scrambled siRNA (NCi) group (P <0.01). The mRNA levels of CD11b and CD80 were decreased in CD36i group compared with NCi group (P <0.01). The cell surface area and cellular adhesiveness were increased in CD36OE group compared with empty vector (vector) group (P <0.05). The mRNA levels of CD11b and CD80 were increased in CD36OE group compared with vector group (P <0.01). The phosphorylation levels of ERK and Src were decreased in CD36i group compared with NCi group (P <0.05). CONCLUSION CD36 promotes the differentiation of human monocyte THP-1 cells to macrophages by increasing the phosphorylation of Src and further activating ERK. 相似文献
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AIM To investigate the role, clinical implications and the underlying mechanisms of Rab11-family interacting protein 4 (Rab11-FIP4) in colorectal cancer (CRC). METHODS The expression levels of Rab11-FIP4 in CRC tissues and corresponding paracancerous tissues were compared by immunohistochemistry, Western blot and RT-qPCR. The above methods were also used to detect the expression levels of Rab11-FIP4 in CRC cells under normal environment and hypoxia. The patiens were divided into Rab11-FIP4 high expression group (n =61) and Rab11-FIP4 low expression group (n =39) according to the immunohistochemical staining score.The overall survival and recurrence time of the 2 groups were compared by Kaplan-Meier survival analysis. HCT116 and LoVo cells with stable over-expression of Rab11-FIP4 were constructed using a lentiviral system. The cytological characteristics effects of Rab11-FIP4 over-expression in CRC cells were examined by CCK-8 assay, clonogenic assay and the Transwell assay. The co-immunoprecipitation was used to detect the correlation between Rab11-FIP4 and insulin-like growth factor 1 receptor (IGF1R). Human phosphokinase array was performed to investigate the signaling pathhway affected by IGF1R in CRC cells with increased expression of Rab11-FIP4. The relationship between Rab11-FIP4 and hypoxia-inducible factor 1α (HIF-1α) was analyzed by tissue microarrays and dual-luciferase reporter assay. RESULTS Rab11-FIP4 expression was up-regulated in CRC tissues and high expression of Rab11-FIP4 was associated with poor prognosis of the patients with CRC (P <0.05). Over-expression of Rab11-FIP4 promoted the viability, migration and invasion of CRC cells (P <0.05). High expression of Rab11-FIP4 regulated ERK1/2 and AKT signaling pathway via IGF1R (P <0.05). Hypoxia promoted the activation of HIF-1α on the Rab11-FIP4 promoter, thereby up-regulating the expression of Rab11-FIP4 (P <0.05). CONCLUSION Rab11-FIP4 may act as an oncogene to regulate migration and invasion of colorectal cancer cells through IGF1R. 相似文献
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AIM To explore the inhibitory effect of metformin （MET） on nerve injury in rats with stroke and its mechanism. METHODS SD rats were randomly divided into sham group （n =15）, model group （n =30）, MET group （n =30）, MET+agomir-NC group （n =30） and MET+agomir group （n =30）. The modified Puisinelli four-vessel occlusion method was used to prepare the model of global ischemic stroke, while the blood vessels in sham rats were isolated without clamping the common artery. One week before modeling, the rats in MET group, MET+agomir-NC group and MET+agomir group were given intraperitoneal injection of 100 mg·kg-1·d-1 MET, 100 mg·kg-1·d-1 MET+40 nmol/d agomir-NC, 100 mg·kg-1·d-1 MET+40 nmol/d miR-29c agomir, respectively, and the rats in sham group and model group were given intraperitoneal injection of the same amount of normal saline. Each treatment in the above groups was given once a day, 0.2 mL each time, for 7 consecutive days. The neurological deficit scores were measured 24, 48 and 72 h after operation. HE staining was used to observe the morphological changes of the hippocampus, and the living neurons were counted. RT-qPCR was used to detect the expression level of miR-29c, and the mRNA levels of silent information regulator 1 （SIRT1） and peroxisome proliferator-activated receptor γ coactivator 1α （PGC-1α） in hippocampus. The protein expression levels of SIRT1 and PGC-1α were determined by Western blot. RESULTS At the same time point, compared with model group, the neurological deficit score in MET group was significantly decreased, and the survival rate of the neurons was significantly increased （P <0.05）. Compared with MET+agomir-NC group, the neurological deficit score in MET+agomir group was increased, and the survival rate of the neurons was significantly decreased （P <0.05）. With the prolongation of time, except for sham group, the neurological deficit score was increased and the survival rate of the neurons was decreased. At 72 h after operation, compared with sham group, the expression of miR-29c in hippocampus of model group was significantly increased, and the mRNA and protein expression levels of SIRT1 and PGC-1α were significantly decreased （P <0.05）. Compared with model group, the expression of miR-29c in hippocampus of MET group was significantly decreased, and the expression of SIRT1 and PGC-1α at mRNA and protein levels was significantly increased （P < 0.05）. Compared with MET+agomir-NC group, the expression of miR-29c in hippocampus of MET+agomir group was significantly increased, and the mRNA and protein expression of SIRT1 and PGC-1α was significantly decreased （P <0.05）. CONCLUSIONS MET alleviates nerve injury in stroke rats, which may be related to down-regulation of miR-29c and promotion of SIRT1/PGC-1α signaling pathway activation. 相似文献
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AIM To investigate the effect of interleukin-33 (IL-33 )-modified bone marrow mesenchymal stem cells (BMSCs) on sepsis-induced acute kidney injury (AKI) in rats and the expression of myeloid differentiation factor 88(MyD88). METHODS A septic rat model was established by cecal ligation and puncture. The SD rats (n =80) were randomly divided into control group, model group, negative transfection group (transplanting untransfected BMSCs) and IL-33 transfection group (transplanting BMSCs transfected with IL-33 ), with 20 in each group. Survival rates of the rats within 72 h in the 4 groups were compared. Serum creatinine (SCr) and blood urea nitrogen (BUN) levels were measured before, and 24, 48 and 72 h after transplantation. The kidney pathological damage was observed by HE staining, and the apoptosis of renal cells was detected by TUNEL method 72 h after transplantation. Western blot was used to detect the protein expression levels of interleukin-1β (IL-1β), IL-6, tumor necrosis factor-α (TNF-α), Toll-like receptor 4 (TLR4) and MyD88. RESULTS The survival rate of the rats in model group was significantly lower than that in control group (P <0.05). The survival rate of the rats in IL-33 transfection group was higher than that in model group and negative transfection group (P <0.05). The levels of SCr and BUN in model group were higher than those in control group (P <0.05). The levels of SCr and BUN in IL-33 transfection group were significantly reduced after transplantation, and were lower than those in model group and negative transfection group (P <0.05). The renal tissue pathological injury score in model group was significantly higher than that in control group (P <0.05). Compared with model group and negative transfection group, the renal tissue pathological injury score in IL-33 transfection group was significantly reduced (P <0.05). The proportion of apoptotic cells in the kidney tissues in model group were higher than that in control group (P <0.05). Compared with model group and negative transfection group, the proportion of apoptotic cells in the kidney tissues in IL-33 transfection group was significantly reduced (P <0.05). The protein expression levels of IL-1β, IL-6, TNF-α, TLR4 and MyD88 in model group were significantly higher than those in control group (P <0.05). Compared with model group and negative transfection group, the protein expression levels of IL-1β, IL-6, TNF-α, TLR4 and MyD88 in IL-33 transfection group were significantly decreased (P <0.05). CONCLUSION IL-33 gene-modified BMSCs significantly improve the renal function of AKI rats with sepsis. The mechanism may be related to IL-33 regulating TLR4/MyD88 signaling pathway and inhibiting renal inflammatory response. 相似文献
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AIM To study the effect of mouse circular RNA-42398 (mmu_circ_42398) over-expression on the activation of hepatic stellate cells. METHODS Mouse hepatic stellate JS1 cells were cultured and randomly divided into control group, vector group and mmu_circ_42398 over-expression group.mmu_circ_42398 over-expression plasmid vector was constructed, and then transiently transfected into JS1 cells using Lipofectamine 2000. The cells were collected 48 h after transfection. Expression of mmu_circ_42398 was detected by RT-qPCR.The backsplice site of PCR products was verified by sequencing. The protein levels of α-smooth muscle actin (α-SMA), collagen type I （Col I), transforming growth factor β1(TGF-β1), Smad2, Smad3, p-Smad2 and p-Smad3 in the cells were determined by Western blot. RESULTS RT-qPCR results showed that the expression of mmu_circ_42398 was significantly increased after mmu_circ_42398 over-expression vector was transiently transfected into the JS1 cells (P <0.01). The protein expression levels of α-SMA and Col I were significantly decreased(P <0.01), and the phosphorylation levels of Smad2 and Smad3 were decreased significantly in mmu_circ_42398 over expression group (P <0.01). However, the protein expression levels of TGF-β1, Smad2 and Smad3 had no significant change (P >0.05). CONCLUSION mmu-circ-42398 inhibits the activation of hepatic stellate cells via TGF-β1/Smads signaling pathway modulation. 相似文献
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AIM To investigate the effect of microRNA-22 (miR-22) secreted by macrophage exosomes on the autophagy of H9c2 cardiomyocytes under uremic toxin stimulation. METHODS The macrophage-derived exosomes stimulated by indoxyl sulfate (IS) were collected and co-cultured with H9c2 cells. The levels of miR-22 in the macrophages, macrophage-derived exosomes and H9c2 cells were detected by RT-qPCR. The viability of H9c2 cells was measured by CCK-8 assay. The expression of exosome surface marker protein CD63 and autophagy-related proteins LC3 and P62 was determined by Western blot. RESULTS Under IS stimulation, the expression of exosome surface marker protein CD63 in the macrophages was significantly higher than that in control group (P <0.05), and the levels of miR-22 in the macrophages and macrophage-derived exosomes were significantly increased (P <0.01). With the increase in macrophage exosome concentration, the viability of H9c2 cells was decreased gradually (P <0.05), and the stimulation of macrophage exosomes reduced P62 expression and promoted the conversion of LC3-I to LC3-II in a dose-dependent manner (P <0.05). Macrophage-derived exosomes increased the ratio of LC3-II to LC3-I but decreased P62 protein expression in the H9c2 cells transfected with miR-22 mimic compared with the cells transfected with corresponding negative control miRNAs (P <0.05). However, miR-22 inhibitor yielded contrasting results. CONCLUSION IS-stimulated macrophages increase expression of miR-22 in cardiomyocytes through exosomes, and promote autophagy of the cardiomyocytes. 相似文献