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AIM: To study the effect of ligustrazine on the cardiacmyocyte lesion in rats with type 2 diabetes mellitus.METHODS: Male Wistar rats were injected with STZ via tail vein under high-glucose and high-fat feeding for 4 weeks to establish the animal model of type 2 diabetes mellitus.Ligustrazine at different doses was used to treat the diabetic rats.The body weight, blood glucose and the morphology of heart tissues were observed.The myocardial levels of IL-1β, IL-6 and TNF-α were detected by ELISA, and the protein expression of IKKβ and NF-κB in the myocardium was determined by Westeren blotting.RESULTS: Ligustrazine at high dose alleviated the body weight reduction and blood glucose elevation cause by diabetes, and reduced pro-inflammatory factors IL-1β, TNF-α and IL-6.Moreover, the protein expression of IKKβ and NF-κB was significant decreased by ligustrazine.CONCLUSION: Ligustrazine inhibits the myocardial inflammation caused by diabetes through anti-inflammatory pathway.
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Aim To study the protective effect of tetra-hydroxystilbeneglucoside ( TSG ) on cardiac injury and the mechanism involved in silent mating type informa-tion regulation 2 homolog 1 ( SIRT1 ) and adenosine monophosphate-activated protein kinase( AMPK) in the diabetic rats. Methods Type 2 diabetic rats were sac-rificed after administration with TSG for 8 weeks. Blood glucose, blood lipids, liverfunction,creatine ki-nase ( CK ) , lactate dehydrogenase ( LDH ) as well as myocardial nonesterified fatty acids( NEFA) were deter-mined by using biochemical test. The concentration of myocardial fatty acid transport proteins ( FATPs ) and-fatty acid β-oxidase ( FA-β-oxidase ) , and the levels of tumor necrosis factor alpha ( TNF-α) , interleukin -6 ( IL-6 ) , interleukin-1β( IL-1β) in serum were also measured by ELISA method and radio immunoassay re-spectively. The protein expressions of TNF-α, IL-6, IL-1β, SIRT1 and AMPK were detected by Western blot. Results Treatment of TSG reduced the contentof blood lipids, NEFA and collagen without affecting the content of blood glucose and insulin. The levels of TNF-α, IL-6 and IL-1βin serum as well as the protein expressions of TNF-α, IL-6 and IL-1β of cardia were also inhibited by administration with TSG. Treatment of TSG caused a significantly increased concentration of myocaidial FATPs and FA-β-oxidase, and dramatically restored the decreased protein expressions of SIRT1 and pAMPK in diabetic rats. Conclusion The protec-tive mechanisms of TSG against diabetic rats are in-volved in the alleviation of inflammatory mediator injury and improving energy metabolism.
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Mesenchymal stem cells (MSCs) are oonsidered as the ideal engineering cells because of their accesibility, their in vitro amplification capacity, low immunogenicity and their tendency toward the tissues. Modification of these cells to express specific anti-cancer molecules or to be a vector for targeted anticancer therapy become a hotspot of biological therapy in recent years. The continual development of MSC study shows great advantage as biological treatment on tumor.
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Objective:To isolate cancer stem cells from cervical carcinoma and to identify their biological characteristics. Methods:Tumor specimens were obtained from 19 cervical cancer patients at stages ⅠA-ⅡB. Primary cells were cultured in tumor sphere medium (TSM) after mechanical dissociation combined with enzymatic digestion. A series of assays were used to identify the characteristics of the sphere forming cells derived from primary culture. Colony formation was observed by limiting dilution method. MTT assay was used to assess proliferation inhibition by paclitaxel and doxorubicin. Cell surface markers were analysed by fluorescence-activated cell sorter (FACS). The expression of stemness-related genes, drug resistance-related genes, and oncogenes were detected by RT-PCR and Western blotting. Tumorigenicity was evaluated by subcutaneous injection of 1×10~5 sphere-forming cells into nude mice. The tumor formation capability was recorded and pathological classification was performed.Results:After 10 to 15 d culture, the formation of non-adherent spheres could be observed in 8 out of 19 primary tumor cells. The formation ratio was increased with the increase in clinical staging. Sphere-forming cells had colony formation capability. Paclitaxel (100 nmol/L) and doxorubicin (100 nmol/L) inhibited the proliferation of these cells by (77.65±6.46)% and (48.00±7.15)%, respectively. The difference was significant (P<0.01). FACS detection results indicated the phenotypes of sphere-forming cells were CD34~-CD105~-CD44~+CK17~+. RT-PCR detection indicated that spheres expressed stemness-related genes (Oct4 and Piwil2), drug resistance gene ABCG2, and oncogenes (c-myc, sox-2 and stat3). Western blotting further indicated stemness-related protein (Oct4 and Piwil2) expression in spheres. Tumors appeared in all animals at 12 weeks after subcutaneous injection of 1×10~5 sphere forming cells and exhibited a high degree of similarity with the primary tumor in cervical cancer patients. Conclusion:Human cervical cancer stem cells were successfully isolated,which provided a useful model for individualized therapy and evaluation of the therapeutic efficacy for cervical cancer patients.
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Objective To investigate the expression and significance of IL-6 and IL-6 receptor (IL-6R)in hysteromyoma.Methods The expression of IL-6 in hysteromyoma and homologous normal myometrium of 20 cases undergoing total hysterectomy were detected by fluorescent quantitative PCR and western-blot,respectively while IL6R was examined by western-blot and RT-PCR.Results The expression of IL-6 protein and mRNA in homologous normal myometrium were obviously higher than that in hysteromyoma[(0.7996 ±0.0359)vs (0.6904 ±0.0414)and (0.0470±0.0071)vs(0.0283±0.0045 ),P＜0.05] ;No difference was observed in IL-6R protein and mRNA expression between leiomyomas and myometrium[(0.7105 ±0.0520)vs (0.6904 ±0.0532)and (0.6644 ±0.0537 )vs (0.6465±0.0501),P＞0.05 ].Conclusion The lower expression of IL-6 in leiomyomas may contribute to the pathogenesis of hysteromyoma to some extent,and there is no relationship between IL-6R and hysteromyoma.
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AIM:To investigate the effect of fluvastatin on the expression of serum and glucocorticoid inducible kinase 1 ( SGK1) and connective tissue growth factor ( CTGF) induced by aldosterone ( Ald) in rat mesangial cells (GMCs). METHODS:GMCs were divided into (1) control group; (2) aldosterone group with different concentrations and times; (3) Ald (10 -7 mol/L) + spironolactone (10 -9 mol/L) group; (4) Ald (10 -7 mol/L) + LY294002 (20 ?mol/L) group; (5) Ald (10-7mol/L) +SB203580 (20 mmol/L) group; (6) the group of Ald (10-7mol/L) + fluvas-tatin at different concentrations (10-7,10-6,10-5 mol/L); (7) Ald (10 -7mol/L) + fluvastatin (10 -5mol/L) + mevalonate (10 -4 mol/L) group; (8) Ald (10 -7 mol/L) + fluvastatin (10 -5 mol/L) + FPP (farnesyl pyrophosphate,10-4 mol/L) group; (9) Ald (10 -7mol/L) + fluvastatin (10 -5 mol/L) + GGPP (geranylgerany pyrophosphate,10 -4 mol/L) group. The protein levels of SGK1 and CTGF were determined by Western blotting. The levels of fibronection (FN),monocyte chemoattractant protein-1 (MCP-1) and intercellular adhesion molecule-1 (ICAM-1) in the supernatants were determined by enzymelinked immunosorbant assay (ELISA). RESULTS:Aldosterone stimulated the protein expression of SGK1 and CTGF in cultured mesangial cells in a dose-dependent manner (P