Lead-induced liver fibrosis and inflammation in mice by the AMPK/MAPKs/NF-κB and STAT3/TGF-ß1/Smad2/3 pathways: the role of Isochlorogenic acid a.

Lead (Pb) is a nonessential heavy metal, which can cause many health problems. Isochlorogenic acid A (ICAA), a phenolic acid present in tea, fruits, vegetables, coffee, plant-based food products, and various medicinal plants, exerts multiple effects, including anti-oxidant, antiviral, anti-inflammatory and antifibrotic functions. Thus, the purpose of our study was to determine if ICAA could prevent Pb-induced hepatotoxicity in ICR mice. An evaluation was performed on oxidative stress, inflammation and fibrosis, and related signaling. The results indicate that ICAA attenuates Pb-induced abnormal liver function. ICAA reduced liver fibrosis, inflammation and oxidative stress caused by Pb. ICAA abated Pb-induced fibrosis and decreased inflammatory cytokines interleukin-1ß (IL-1ß) and tumor necrosis factor-alpha (TNF-α). ICAA abrogated reductions in activities of superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx). Masson staining revealed that ICAA reduced collagen fiber deposition in Pb-induced fibrotic livers. Western blot and immunohistochemistry analyses showed ICAA increased phosphorylated AMP-activated protein kinase (p-AMPK) expression. ICAA also reduced the expression of collagen I, α-smooth muscle actin (α-SMA), phosphorylated extracellular signal-regulated kinase (p-ERK), phosphorylated c-jun N-terminal kinase (p-JNK), p-p38, phosphorylated signal transducer and phosphorylated activator of transcription 3 (p-STAT3), transforming growth factor ß1 (TGF-ß1), and p-Smad2/3 in livers of mice. Overall, ICAA ameliorates Pb-induced hepatitis and fibrosis by inhibiting the AMPK/MAPKs/NF-κB and STAT3/TGF-ß1/Smad2/3 pathways.

Prevention of polycystic ovary syndrome and postmenopausal osteoporosis by inhibiting apoptosis with Shenling Baizhu powder compound.

Objective: Shenling Baizhu powder (SBP) has been shown to reverse the abnormal expression of the aromatic hydrocarbon receptor (AHR) mediated by air pollution. Our study aimed to understand the main ingredient of SBP and investigate its action mechanism in preventing polycystic ovary syndrome (POCS) and postmenopausal osteoporosis (PMO). Methods: The active ingredients of SBP with the highest binding affinity to AHR were screened using a Chinese medicine database, and their binding mechanism was simulated using molecular dynamics simulation (MDS). Rutin was utilized to treat ovarian granulosa cell lines and osteoblast cell lines. The cell lines were treated with a gradient of rutin concentration (0.01 mmol/L, 0.05 mmol/L and 0.1 mmol/L) to find the optimal drug dose. PCR was used to detect AHR and apoptosis-related proteins, and WB to detect the expression of AHR, caspase-3 and cleaved-caspase-3. Finally, the CCK-8 cell proliferation assay detected the proliferation of cells. Results: We obtained Rutin through the Chinese medicine database, and dynamics simulation determined its binding sites. Ovarian granulosa cell lines and osteoblast cell lines were treated with Rutin. RT-PCR and western blotting revealed that the expression of apoptosis-associated protein Bcl-2 was elevated, and the expression of AHR, Bax, caspase-3 and PARP were decreased. CCK-8 results showed accelerated proliferation in both cell types. Conclusion: Rutin, the main ingredient of SBP compound, works by binding to AHR, which can improve POCS and PMO by inhibiting cell apoptosis and by promoting cell proliferation.

Therapeutic effect and potential mechanisms of intra-articular injections of miR-140-5p on early-stage osteoarthritis in rats.

MicroRNAs (miRs) receive extensive attention in osteoarthritis (OA) pathogenesis in recent years, and our previous study confirmed that an intra-articular injection (IAJ) of miR-140-5p alleviates early-stage OA (EOA) progression in rats. This study aims to investigate the therapeutic effect and potential mechanisms of single IAJ (SIAJ) of miR-140-5p on different stage OA and multiple IAJs (MIAJ) of miR-140-5p on EOA. Firstly, the OA model was surgically induced in rats, nine were treated with IAJ of Cy5-miR-140-5p at one week after surgery, and fluorescence distribution was analyzed at different times. Then, 72 rats were treated with SIAJ of miR-140-5p at different stages or MIAJ of miR-140-5p at one week after surgery, and OA progression was evaluated macroscopically and histologically at different times. Finally, the downstream targets and underlying molecular mechanisms of miR-140-5p were predicted by bioinformatics and partially validated. As a result, the intra-articularly injected miR-140-5p entered cartilage and could be taken up by chondrocytes rapidly. IAJ(s) of miR-140-5p improved the behavioral scores, chondrocyte number, cartilage thickness, and pathological scores to varying degrees. Specifically, the earlier a SIAJ of miR-140-5p was administrated, the better the therapeutic effect; meanwhile, MIAJ of miR-140-5p exhibited a better therapeutic effect than SIAJ on EOA. Eighty-four potential target genes and mechanisms of rno-miR-140-5p were predicted, and the effect of miR-140-5p on the potential target genes VEGFA and JAG1 was experimentally validated. Collectively, IAJs of miR-140-5p effectively alleviate EOA progression by modulating multiple biological processes and pathways in rats, representing a promising therapeutic for EOA.

TLR3 inhibitor and tyrosine kinase inhibitor attenuate cigarette smoke/poly I:C-induced airway inflammation and remodeling by the EGFR/TLR3/MAPK signaling pathway.

Tobacco smoke is the major risk factor for developing chronic obstructive pulmonary disease (COPD). Viral infection is a major cause of COPD exacerbation, which lacks effective drug treatments. In the present study, to mimic the pathogenesis of COPD, we employed a TLR3 ligand [Poly (I:C), PIC] to mimic viral infection to determine whether it enhances the effects of cigarette smoke (CS)-induced airway inflammation and remodeling. Our results showed that PIC enhanced the effects of cigarette smoke extract (CSE)-induced inflammatory cytokine IL-1ß, TNF-α and IL-8 mRNA expression and remodeling factor E-cadherin, α-SMA and TGF-ß1 mRNA expression with TLR3 upregulation and EGFR phosphorylation in pulmonary epithelial NCI-H292 cells. These responses were inhibited by a TLR3/dsRNA complex inhibitor (TLR3i) or a tyrosine kinase inhibitor icotinib (Ico). Similarly, in the PIC-enhanced CS-induced airway inflammation and remodeling mouse model, treatment with TLR3i or Ico reduced the mRNA and protein expression of the inflammatory cytokines IL-1ß and TNF-α and keratinocyte chemoattractant (KC) and the remodeling factors α-SMA, TGF-ß1, MMP-9 and MUC5AC, while increasing E-cadherin mRNA and protein expression. Furthermore, we found that TLRi and Ico can attenuate the airway hyperreactivity induced by PIC, which is enhanced by CS. Finally, PIC enhanced the effects of CS on TLR3 upregulation and EGFR phosphorylation and significantly increased Erk1/2 and P38 phosphorylation, whereas TLR3i and Ico markedly suppressed TLR3 upregulation and EGFR, Erk1/2 and P38 phosphorylation in the model. Our findings suggest that TLR3/EGFR may be a potential target for the treatment of airway inflammation and remodeling in COPD.

Bone marrow-derived mesenchymal stem cells repair severe acute pancreatitis by secreting miR-181a-5p to target PTEN/Akt/TGF-ß1 signaling.

BACKGROUND: Severe acute pancreatitis (SAP) is associated with high morbidity and mortality. Bone marrow mesenchymal stem cells (BMSCs) have shown obvious protective effect on SAP. However, little is known about the underlying mechanism. The objective of this study is to unravel the role and regulatory mechanism of miR-181a-5p in BMSCs-mediated pancreatic repair. METHODS: BMSCs were isolated from Sprague-Dawley rats and characterized by flow cytometry and Oil Red O staining. Sodium taurocholate- and caerulein-induced models were used as SAP models in vivo and in vitro, respectively. Pancreatic injury were evaluated by H&E and histopathological analysis, as well as by measuring levels of amylase, lipase and cytokines. qRT-PCR and western blotting were performed to detect the level of miR-181a-5p and the protein levels of PTEN/Akt, respectively. ELISA was conducted to detect the levels of TNF-α, IL-1ß, IL-6, angiopoietin, IL-4, IL-10 and TGF-ß1. The apoptotic rate of AR42J cells was quantitated by concurrent staining with Annexin-V-FITC and PI. RESULTS: BMSCs significantly attenuated pancreatic injury in SAP rats by reducing inflammatory infiltration and necrosis, and this effect was abolished by CXCR4 agonist AMD3100. ADM3100 exhibited more severe pancreatic injury and decreased miR-181a-5p levels in the pancreas and serum compared to SAP group. Overexpression of miR-181a-5p in BMSCs (BMSCs-miR-181a-5p) markedly potentiated the protective effect of BMSCs by reducing histological damage and levels of amylase and lipase. Moreover, BMSCs-miR-181a-5p dramatically reduced levels of angiopoietin, TNF-α, IL-1ß and IL-6, but induced the levels of IL-4 and IL-10. In caerulein-treated AR42J cells, co-culturing of BMSCs-miR-181a-5p alleviated caerulein-induced increase of amylase and lipase, and apoptosis via PTEN/Akt/TGF-ß1 signaling. CONCLUSION: BMSCs alleviate SAP and reduce inflammatory responses and apoptosis by secreting miR-181a-5p to target PTEN/Akt/TGF-ß1 signaling. Hence, BMSCs-miR-181a-5p could serve as potential therapeutic target for SAP.

[Association of Serum Soluble CD36 with Clinical Variables in Diabetic Patients with Chronic Kidney Diseas].

OBJECTIVE: To investigate the levels of serum soluble CD36 (sCD36) in patients of diabetes mellitus (DM) with chronic kidney disease (CKD),and to analyze its correlation with clinical indicators. METHODS: A total of 161 patients with CKD were enrolled in this study. The patients were divided into two groups according to whether they had DM or not: DM+CKD group and non-DM CKD group. The levels of carotid intima-media thickness (IMT) and the combination of atherosclerotic plaques were measured by color Doppler ultrasonography. Serum fasting serum samples were collected and serum sCD36 level was measured by ELISA. the status of serum sCD36 was analyzed with the progress of renal disease,and the correlation of sCD36 level with clinical indicators were analyzed. RESULTS: Among the 161 patients,87 (54%) were DM+CKD and 74 (46%) were non-DM CKD. There was no significant difference in the levels of urea nitrogen (BUN),serum creatinine (sCr),estimated glomerular filtration rate (eGFR),cystatin C (Cys-C),triglyceride (TG),cholesterol (Chol),low density lipoprotein-chol (LDL-C),urinary albumin/creatinine and IMT in the two groups (P>0.05). Compared with non-DM CKD group,the serum sCD36 level (U/L) in DM+CKD group was lower (4.58±1.06 vs. 4.97±1.28,P<0.05). In DM+CKD group,serum sCD36 was negatively correlated with BUN,sCr and Cys-C (r=⁻0.355,⁻0.336,⁻0.323; P<0.01),and positively correlated with eGFR (r= 0.399; P<0.01),but not with TG,Chol,LDL-C or IMT (P>0.05). In non-DM CKD group,there was a positive correlation between sCD36 and TG,Chol and LDL-C (r= 0.251, 0.298, 0.292; P<0.05),and negatively correlated with Cys-C (r=⁻0.287; P<0.05),but not with eGFR,BUN,sCr or IMT (P>0.05). With the progress of CKD,serum sCD36 levels gradually decreased (P>0.05). CONCLUSION: Serum sCD36 level is associated with renal function in the patients with DM complicated with CKD,but not with lipid indicators.

[Visual field analysis after laser in situ keratomileusis].

OBJECTIVE: To evaluate the effects of laser in situ keratomileusis (LASIK) on the patients' visual field. METHODS: 92 eyes of 46 patients with mild, moderate, and high degrees of myopia were undergone LASIK. Perimetry was performed before and 1 day, 1 month, 3 months, 6 months after LASIK surgery by Humphrey II-750 visual field analysis. The central 30 - 2 fast threshold program was used to examine the change in central 30 degree. Mean defect (MD) and pattern standard deviation (PSD) were record. All operations were performed by an experienced surgeon. Statistical analyses were performed by SPSS using the randomized complete block design ANOVA (two factors analysis of variance). Differences among variables were evaluated by q-test (Student-Newman-Kueuls method). RESULTS: 93.5% patients obtained estimated best correct visual acuity or better at six months after LASIK. No complications were found in the subjects. No difference was found in fundus examinations before and after surgery. Intraocular pressure was normal at all preoperative and postoperative examinations. MD was significant differences among preoperative and postoperative values (P < 0.05, for all comparisons) and was correlated with visual acuity post-LASIK significantly (P < 0.05). While PSD did not show significant differences before and after LASIK (P > 0.05) and was not correlated with visual acuity post-LASIK. The indexes of visual field were not correlated with the degree of myopia and the other surgical parameters (P > 0.05). CONCLUSION: In current research, no significant visual field change was demonstrated pre-LASIK and post-LASIK after 6 months follow-up. The study suggests that LASIK does not affect the retinal physiological functions.