1.Migration and invasion abilities of normal fibroblast-like synoviocytes versus fibroblast-like synoviocytes in rheumatoid arthritis
Jun HU ; Zizheng TANG ; Yu ZHANG ; Weigan SHEN ; Guiping LIU ; Wenwen WANG ; Jiahuan LIU
Chinese Journal of Tissue Engineering Research 2017;21(24):3870-3874
BACKGROUND:Fibroblast-like synoviocytes (FLS) in the synovial lining layer are related to the cell proliferation, invasion, migration and apoptosis as well as bone resorption in rheumatoid arthritis. OBJECTIVE:To compare the migration and invasion abilities of FLS (MH7A) in rheumatoid arthritis and normal FLS (HFLS). METHODS:The capacities of cell migration and invasion were evaluated by Transwell cell migration and invasion assays. The primers of the indicated microRNAs were designed and synthesized, and the expression levels of miRNAs were determined by real-time PCR according to the SYBR?PrimeScript?miRNA RT-PCR Kit instruction. RESULTS AND CONCLUSION:MH7A possessed stronger migration and invasion abilities than HFLS. Compared with HFLS, obviously upregulated miR-132, -155, -203, -223 and -124, and significantly downregulated miR-15a, -16, 18a, -19a, -26a and -146a were found in MH7A. These findings suggest that the differentially expressed 11 kinds of rheumatoid arthritis-associated miRNAs participate in the pathogenesis of rheumatoid arthritis probably by enhancing the migration and invasion capacities of MH7A.
2.The influence of chronic intermittent hypoxia on hypothalamic-somatotropic axis in rats
Jiahuan XU ; Hui SHEN ; Wenyang LI ; Yanling MENG ; Wei WANG
Chinese Journal of Internal Medicine 2019;58(8):592-595
Objective To investigate the effects of chronic intermittent hypoxia on somatotropic axis hormone levels in rats.Methods Mature male Wistar rats were exposed to air or intermittent hypoxia randomly.The serum levels of growth hormone-releasing hormone (GHRH),growth hormone (GH) and somatostatin (SS) were measured before exposure,at the 4th,8th,and 12th week after exposure.Different hormone levels in two groups were compared and analyzed.Results Compared with the control group,GHRH levels in chronic intermittent hypoxic group showed a significant decline at the 4th week [(732.77± 46.99)pg/ml vs.(893.59±40.00) pg/ml,P<0.05],while SS levels at the 8th week [(30.71 ±2.27) pg/ml vs.(44.69±3.36) pg/ml,P<0.05] and GH levels at the 12th week [(1.20±0.29) ng/ml vs.(2.06±0.13) ng/ml,P<0.05]were similarly reduced.As the duration of intermittent hypoxia was prolonged,the GHRH levels did not decrease further [4th week (732.77±46.99) pg/ml vs.8th week (607.54± 131.61) pg/ml vs.12th week (730.05±40.63) pg/ml,P>0.05].However,the serum SS levels decreased further from the 8th week to the 12th week [(30.71±2.27) pg/ml vs.(24.41±4.06) pg/ml,P<0.05].Conclusion Chronic intermittent hypoxia might inhibit the function of somatotropic axis.Hypothalamic hormones are the earlyonesto be influenced,thereafter the entire axis.
3.The influence of excessive daytime sleepiness and sleep quality on anxiety and depression in patients with obstructive sleep apnea hypopnea syndrome
Qi QI ; Wei WANG ; Hui SHEN ; Zheng QIN ; Ling WANG ; Jiahuan XU ; Wenyang LI ; Weiyi QI ; Jian KANG
Chinese Journal of Internal Medicine 2019;58(2):119-124
Objective To explore the factors that contribute to the anxiety and depression in obstructive sleep apnea hypopnea syndrome (OSAHS) patients in terms of excessive daytime sleepiness (EDS) and sleep quality.Methods A total of 196 OSAHS patients,including 103 severe patients and 93 mild-moderate patients,were enrolled.Polysomnography was carried on at the sleep center of the First Hospital of China Medical University between May 2013 and November 2015.According to the Epworth sleepiness scale (ESS) and the subject daytime sleepiness symptom,all patients were divided into EDS group and non-EDS group.The patients' general information and subjective symptoms were recorded.Emotional states were assessed with self-rating anxiety scale (SAS) and self-rating depression scale (SDS).Sleep quality was evaluated with Pittsburgh sleep quality index (PSQI).The anxiety and depression related factors were studied by regression analysis.Results (1) In severe OSAHS group,the patients with EDS showed higher PSQI(6.22± 2.57 vs.4.05± 3.72,P<0.01) and oxygen desaturation index(ODI)[(57.70±17.53) events/h vs.(48.23 ± 22.01)events/h,P<0.05] when compared with those without EDS.(2) In both severe and mild-moderate OSAHS groups,the patients with EDS presented higher SAS scores (severe:33.86±7.60 vs.28.95 ± 4.71,mild-moderate:37.46± 10.68 vs.33.40± 11.07,P<0.05)and SDS scores(severe:32.81 ± 8.36 vs.28.90±4.53,mild-moderate:36.98± 12.77 vs.31.70±10.94,P<0.05)when compared with those without EDS.(3) The multiple regression analysis showed that the SAS scores were related to ESS,PSQI,insomnia and nasal obstruction (R2=0.356,P<0.05),and the SDS scores were related to ESS,PSQI and insomnia(R2=0.344,P<0.05).Conclusions The anxiety and depression of OSAHS patients are closely related to the severity of EDS and sleep quality.Both severe and mild-moderate OSAHS patients with EDS have worse anxiety and depression scores.
4.Macrophage migration inhibitory factor protects bone marrow mesenchymal stem cells from hypoxia/ischemia-induced apoptosis by regulating lncRNA MEG3.
Zhibiao BAI ; Kai HU ; Jiahuan YU ; Yizhe SHEN ; Chun CHEN
Journal of Zhejiang University. Science. B 2022;23(12):989-1001
OBJECTIVES:
This research was performed to explore the effect of macrophage migration inhibitory factor (MIF) on the apoptosis of bone marrow mesenchymal stem cells (BMSCs) in ischemia and hypoxia environments.
METHODS:
The cell viability of BMSCs incubated under hypoxia/ischemia (H/I) conditions with or without pretreatment with MIF or triglycidyl isocyanurate (TGIC) was detected using cell counting kit-8 (CCK-8) analysis. Plasmids containing long noncoding RNA (lncRNA) maternally expressed gene 3 (MEG3) or β-catenin small interfering RNA (siRNA) were used to overexpress or downregulate the corresponding gene, and the p53 signaling pathway was activated by pretreatment with TGIC. The influences of MIF, overexpression of lncRNA MEG3, activation of the p53 signaling pathway, and silencing of β-catenin on H/I-induced apoptosis of BMSCs were revealed by western blotting, flow cytometry, and terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick-end labeling (TUNEL) staining.
RESULTS:
From the results of CCK-8 assay, western blotting, and flow cytometry, pretreatment with MIF significantly decreased the H/I-induced apoptosis of BMSCs. This effect was inhibited when lncRNA MEG3 was overexpressed by plasmids containing MEG3. The p53 signaling pathway was activated by TGIC, and β-catenin was silenced by siRNA. From western blot results, the expression levels of β-catenin in the nucleus and phosphorylated p53 (p-p53) were downregulated and upregulated, respectively, when the lncRNA MEG3 was overexpressed. Through flow cytometry, MIF was also shown to significantly alleviate the increased reactive oxygen species (ROS) level of BMSCs caused by H/I.
CONCLUSIONS
In summary, we conclude that MIF protected BMSCs from H/I-induced apoptosis by downregulating the lncRNA MEG3/p53 signaling pathway, activating the Wnt/β-catenin signaling pathway, and decreasing ROS levels.
Humans
;
RNA, Long Noncoding/metabolism*
;
Macrophage Migration-Inhibitory Factors/metabolism*
;
beta Catenin/metabolism*
;
Reactive Oxygen Species/metabolism*
;
Sincalide/metabolism*
;
Tumor Suppressor Protein p53/metabolism*
;
Apoptosis
;
Mesenchymal Stem Cells
;
Wnt Signaling Pathway/genetics*
;
RNA, Small Interfering/metabolism*
;
Hypoxia/metabolism*
;
Ischemia
;
Bone Marrow Cells
5.Study on Improvement Effects and Mechanism of Paeoniflorin on Myocardial Injury in Type 2 Diabetic Model Rats
Shanshan LI ; Chunyu TIAN ; Guowei ZHANG ; Leilei MA ; Ji’an LI ; Jiaxuan LIU ; Ying LIU ; Shuang LI ; Xiaolong LI ; Jiahuan SHEN
China Pharmacy 2021;32(23):2846-2853
OBJECTIVE:To study the improvement effects of paeon iflorin(PF)on myocardial injury in type 2 diabetes mellitus(T2DM)model rats and its mechanism. METHODS :The experiment was set up in the normal group ,model group , positive control group (metformin 90 mg/kg),PF high-dose ,medium-dose and low-dose groups (90,60,30 mg/kg),with 8 rats in each group. Except for normal group ,other groups were given high-glucose and high-fat diet and intraperitoneal injection of streptozotocin (30 mg/kg) to induce T 2DM model. After modeling , administration groups were given relevant medicine intragastrically,normal group and model group were given constant volume of normal saline intragastrically ,once a day ,for consecutive 4 weeks. The body weight ,fasting blood glucose and oral glucose tolerance were measured ;serum levels of glycosylated serum protein (GSP),total cholesterol (TC),triacylglycerol(TG),glutathione peroxidase (GSH-Px),superoxide dismutase(SOD),malondialdehyde(MDA),creatine kinase isoenzyme-MB (CK-MB) and troponin Ⅰ (cTn Ⅰ) were determined. The pathomorphological changes of myocardium were observed. The apoptosis index of rat cardiomyocytes was ( detected. The protein expression of B-cell lymphoma 2 (Bcl-2),Bcl-2 related X protein (Bax)and caspase- 3 in rat myocardium were detected by immunohistochemistry and Western blot. RE SULTS:Compared with normal group ,the body weight ,serum levels of GSH-Px and SOD ,protein expression of Bcl- 2 in myocardium were decreased significantly in model group(P<0.01);while fasting blood glucose ,area under blood glucose curve ,serum levels of biochemical indexes (GSP,TC, TG,MDA,CK-MB,cTnⅠ),cardiomyocyte apoptosis index ,protein expression of Bax and caspase- 3 in myocardium were increased significantly (P<0.05 or P<0.01). The arrangement of myocardium was relatively irregular ,and some muscle fibers were broken. Compared with model group ,except for body weight ,serum levels of SOD and MDA ,the protein expression of Bax in myocardium in PF low-dose group , above indexes of PF groups were reversed significantly (P<0.05 or P<0.01). CONCLUSIONS:PF can regulate glycolipid metabolism ,enhance antioxidant ability ,inhibit cardiomyocyte apoptosis and improve myocardial injury in T 2DM model rats ;the mechanism may be associated with increasing the protein expression of Bcl- 2 and down-regulating the protein expression of Bax and caspase- 3 in myocardium.