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Aim To research the effect of gender on immune function and keratinocyte damage in imiquimod(IMQ)induced psoriasis mice.Methods IMQ-induced psoriasis mice were freely divided into female and male model groups, and female and male normal groups were set up smeared with an equal amount of petroleum.PASI score and HE staining were used to evaluate skin lesion and pathology; Western blot and immunity fluorescence were used to detect the expression of proliferating cell nuclear antigen(PCNA), Ki67, keratin 1(K1), keratin 10(K10), and involucrin in skin lesions; spleen index of mice was calculated; immunohistochemistry was used to detect the expression of CD4, IFN-γ, IL-4, and IL-17 in skin and spleen; flow cytometry was used to detect the changes of Th1 and Th17 cell subsets in spleen.Results Compared with female and male normal groups, PASI score of female and male model groups increased, skin lesions were abnormally thickened and differentiated, the level of PCNA and Ki67, the spleen index, the Th1 and Th17 cell subsets in the spleen both increased, K1, K10, and involucrin decreased, the levels of CD4, IFN-γ, IL-17 in skin lesions and spleen were elevated, but the level of IL-4 showed the opposite trend.There was no statistical difference in the above indicators between the female and male model groups.Conclusion Gender has no effect on the abnormal activation of T cell immune function and the proliferation and differentiation of keratinocytes in psoriasis-like mice induced by IMQ.
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BACKGROUND: Traditional Chinese medicine has certain value and significance in the treatment of ischemic cardiovascular and cerebrovascular diseases by regulating the ischemia-hypoxia microenvironment and improving the survival rate and differentiation rate of stem cells. OBJECTIVE: To sort out and analyze the research progress of traditional Chinese medicine on regulating ischemia-hypoxia microenvironment intervention on proliferation, differentiation, aging and autophagy of bone marrow mesenchymal stem cells in recent years. METHODS: The full-text database of Chinese journals, PubMed and Wanfang were retrieved with the keywords of “bone mesenchymal stem cells, ischemia-hypoxia microenvironment, proliferation, differentiation, aging” in English or “bone marrow mesenchymal stem cells, ischemia and hypoxia, proliferation, differentiation, aging” in Chinese for articles regarding effects of ischemia and hypoxia microenvironment on survival rate and differentiation rate of bone marrow mesenchymal stem cells published from 2002 to 2019. Fifty-five articles were selected for review, including 22 Chinese articles and 33 English articles. RESULTS AND CONCLUSION: The ischemia-hypoxia microenvironment is the important reason for the low survival rate and differentiation rate of bone marrow mesenchymal stem cells. There are many adverse reactions in the intervention of bone marrow mesenchymal stem cells with gene modification or cell molecules and drugs, which have become difficult problems to be solved in modern medicine. Exploring the internal relationship between microenvironment and stem cells using single or active components of traditional Chinese medicine combined with RNA transcriptomics is a new way to improve the viability of stem cells.
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BACKGROUND: Tissue engineering technology has emerged to solve the repair and reconstruction of tissues and organs. The selection and application of seed cells become an issue of concern. Fibroblasts are a popular choice in various tissue engineering studies. OBJECTIVE: To summarize the biological characteristics of fibroblasts, their multi-potential differentiation and their effects on the proliferation and differentiation of stem cells. METHODS: A computer-based research of CNKI (2015-2019) and PubMed (2005-2019) databases was performed for the articles about the biological characteristics of fibroblasts and multipotential differentiation. The articles were systematically summarized and analyzed. The newest research process of multi-potential differentiation of fibroblasts was reviewed. RESULTS AND CONCLUSION: Fibroblasts hold strong metabolism and proliferation abilities, which can synthesize and secrete protein. Fibroblasts can differentiate into different cells under different environments, and exhibit the same strong multi-potential differentiation with stem cells. Therefore, it is commonly used in the transdifferentiation, cell culture, injury repair and tissue engineering, and their effects on promoting proliferation and inducing differentiation of stem cells are particularly significant. We should fully utilize the biological characteristics and multi-potential differentiation of fibroblasts, and co-culture of fibroblasts with stem cells can provide seed cells for tissue engineering, which further provide an idea for traumatic repair in clinic.
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BACKGROUND: In the treatment of bone defect or bone injury by tissue engineering, biomaterials affect the survival rate, proliferation and differentiation of bone marrow mesenchymal stem cells. OBJECTIVE: To review the research progress regarding how biomaterials affect the proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells, and to guide the rational application of biomaterials. METHODS: A computer-based online search of CNKI, PubMed, Web of Science, and Wanfang database with the search terms “Bone marrow mesenchymal stem cells, Osteogenic differentiation, Biological materials, The microstructure”. The eligible literatures regarding the effects of different biomaterials on the proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells were included. RESULTS AND CONCLUSION: (1) Metallic materials have the advantages of good biocompatibility, bone conductivity, and mechanical performance. Non-metallic materials exhibit good biocompatibility, bone conductivity, reabsorption, and three-dimensional shaping. (2) There are many factors that affect the surface microstructure of the biomaterial. As for the same biomaterial, greater surface energy/ wettability leads to better proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells; greater roughness better promotes the proliferation, adhesion, and differentiation of bone marrow mesenchymal stem cells; larger pore diameter and lower pore diameter rate are more prone to promote the osteogenic differentiation of bone marrow mesenchymal stem cells; greater substrate rigidity and elastic modulus better facilitate the osteogenic differentiation of bone marrow mesenchymal stem cells. The abovementioned factors of the biomaterials affect the proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells. These findings help promote the application of biomaterials seeded with bone marrow mesenchymal stem cells in the clinic.
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BACKGROUND: Alendronate (AL), a drug for inhibiting osteoclast-mediated bone-resorption, was intercalated into an inorganic drug delivery nanovehicle, layered double hydroxide (LDH), to form a new nanohybrid, AL-LDH, with 1:1 heterostructure along the crystallographic C-axis. Based on the intercalation reaction strategy, the present AL-LDH drug delivery system (DDS) was realized with an enhanced drug efficacy of AL, which was confirmed by the improved proliferation and osteogenic differentiation of osteoblast-like cells (MG63). METHODS: The AL-LDH nanohybrid was synthesized by conventional ion-exchange reaction and characterized by powder X-ray diffraction (PXRD), high-resolution transmission electron microscopy (HR-TEM), and Fourier transform infrared (FT-IR) spectroscopy. Additionally, in vitro efficacy tests, such as cell proliferation and alkaline phosphatase (ALP) activity, were analyzed. RESULTS: The AL was successfully intercalated into LDH via ion-exchange reaction, and thus prepared AL-LDH DDS was X-ray single phasic and chemically well defined. The accumulated AL content in MG63 cells treated with the AL-LDH DDS nanoparticles was determined to be 10.6-fold higher than that within those treated with the intact AL after incubation for 1 hour, suggesting that intercellular permeation of AL was facilitated thanks to the hybridization with drug delivery vehicle, LDH. Furthermore, both in vitro proliferation level and ALP activity of MG63 treated with the present hybrid drug, AL-LDH, were found to be much more enhanced than those treated with the intact AL. This is surely due to the fact that LDH could deliver AL drug very efficiently, although LDH itself does not show any effect on proliferation and osteogenic differentiation of MG63 cells. CONCLUSION: The present AL-LDH could be considered as a promising DDS for improving efficacy of AL.
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Alendronate , Alkaline Phosphatase , Cell Proliferation , Drug Delivery Systems , Fourier Analysis , In Vitro Techniques , Microscopy, Electron, Transmission , Nanoparticles , Spectrum Analysis , X-Ray DiffractionABSTRACT
Objective To investigate the effect of maternal separation (MS) on cognitive function in adult male rats through the expression of neuronal nitric oxide synthase (nNOS) in hippocampus, and to reveal the roles of early life stress (ELS) on neural development in rats. Methods Healthy SD pregnant rats (n=12) were randomly divided into maternal separation group (MS group) and control group (NMS group) (n=6 for each group). The newborn rats in the MS group were separated from the mother rats for 3 h every day from postnatal day 3 to 22 whereas no intervention was taken in the NMS group. At the age of 10 weeks, Morris water maze was used to test the learning and memory abilities of two groups of offspring male rats. Neuron immunofluorescence staining was used to examine the number and distribution of neurons in dentate gyrus (DG) of two groups of offspring male rats. Western Blot method was used to detect nNOS, eNOS, Bax/BCL2, Caspase-3 and P53 levels in the hippocampus of the two groups. Ki67/DCX immunofluorescence staining were used to examine the proliferation and differentiation of neurons in the DG area of the hippocampus. TUNEL staining was used to detect the neuronal degeneration and death in the DG area of the hippocampus. Results Behavioral tests showed that the escape latency of male rats in MS group was prolonged, the target quadrant residence time and the number of platform crossing decreased (P<0.05) compared with NMS group. Compared with NMS group, the number of normal and degenerated neurons in hippocampal DG area of MS group had no significant change (P＞0.05). However, the expression of nNOS and eNOS in hippocampus was decreased (P<0.05) and the expression of Bax/BCL2 was increased (P<0.05), but the expression of caspase-3 and P53 remained unchanged (P＞0.05). In addition, Neuronal proliferation and differentiation were decreased and apoptosis was increased in MS group (P<0.05). Conclusion Repeated MS reduces the expression levels of nNOS in the hippocampus, affects the neuronal function in the DG area, and has a long-term influence on the neurodevelopment, which results in cognitive deficits related to learning and memory abilities in adult rats.
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Objective To explore the influences of proliferation and differentiation of preadipocytes on high fat diet-induced obesity and obesity prevention through exercises in rats.Methods Forty male Sprague-Dawley rats were divided into a normal diet control group (C,n=8),a normal diet with exercises group (E,n=8),a high fat diet control group (H,n=14) and a high fat diet with exercises group (HE,n=10).Group C and group H kept sedentary,while group E and group HE underwent treadmill exercises at about 75％VO2max level.After 12 weeks of intervention,the body weight,epididymal,perirenal and retroperitoneal fat mass were recorded,and the total fat mass was determinated.Stereology method was used to calculate the number and average diameter of fat cells.Western Blotting was conducted to measure the expression of PPARγ and C/EBPα protein in adipose tissues.Results (1) The body weight and total fat mass of group H were significantly higher than those of group C (P＜0.01).Perirenal,retroperitoneal fat and total fat mass of group E and HE were significantly lower than those of group C and H respectively (P＜0.01).(2) The total fat cell number of group H was significantly higher than that of group C.The average diameter of fat cells in perirenal and retroperitoneal fat pads of group E were significantly lower than that of group C (P＜0.05,P＜0.01),while that of group HE in epididymal,perirenal and retroperitoneal fat pads was significantly lower than that of group H (P＜0.01).(3) Compared with group C,the expression of PPARγ protein of group E and H increased significantly in epididymal,perirenal and retroperitoneal fat pads (P＜0.01).The expression of C/EBPα protein of group H was significantly higher than that of group C in epididymal and perirenal fat pads (P＜0.01),the expression of C/EBPα protein of group E was significantly higher than that of group C in perirenal and retroperitoneal fat pads (P＜0.01),while the expression of C/EBPα protein of group HE was significantly lower than that of group H in perirenal fat pads(P＜0.01).Conclusion The proliferation and differentiation of preadipocyte was enhanced in the development of high fat diet induced obesity and exercises to prevent obesity,but its role and mechanisms were different.The high fat diet increases the number of fat cells which is a compensatory mechanism for the body to adapt to fat accumulation,while exercises might promote cell renewal and decrease the average size of fat cells which is an adaptive mechanism to improve fat storage.
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Objective To study the effects of Yunnan BaiYao on the expressions of c-fos and c-jun mRNA in HPDLFs and explore the potential mechanism that Yunnan BaiYao promotes the proliferation and differentiation of HPDLFs.Methods The HPDLFs tissue was obtained from the extracted healthy premolar.The HPDLFs used underwent four to six passages.Cells were divided into untreated group,positive control group and treated group.In treated group,HPDLFs were co-cultured with Yunnan BaiYao solution for 4h with gradient concentration.The expressions of c-fos and c-jun mRNA were determined by RT-PCR.Results The results showed that the expressions of c-fos and c-jun mRNA were up-regulated significantly in treated group compared with control group (P＜0.05) Conclusion Yunnna BaiYao can upregulate the expressions of c-los and C-jun mRNA in HPDLFs.Through upregulating the expressions of c-fos and c-jun mRNA in hPDLs,Yunnan BaiYao can promote proliferation and differentiation of ossification of HPDLFs to induce bone formation.
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Objective To investigate the effect of 1,25-(OH)2-vitamin D3 (VD3) or mechanical strain alone and their combined treatment on proliferation and differentiation of pre-osteoblast MC3T3-E1 cells in vitro, as well as gene and protein expression of osteoprotegerin (OPG) and receptor activator of nuclear factor-кB ligand (RANKL) in those cells. Methods MC3T3-E1 cells were treated with 10 nmol/L VD3, intermitted mechanical strain or with a combination of these two factors. Cell proliferation was assessed with flow cytometry, and alkaline phosphatase (ALP) activity was measured using a fluorometric detection kit. The mRNA expression of ALP, runt-related transcriptional factor 2 (Runx2), OPG, and RANKL genes was determined by real-time PCR. The proteins expression of Runx2, OPG, and RANKL was determined by Western blotting. ResultsVD3 inhibited the proliferation of MC3T3-E1 cells, but the mechanical strain had no effect on cell proliferation. Mechanical strain, VD3, and the combined treatment enhanced the ALP activity of MC3T3-E1 cells as well as the protein expression of Runx2. The effect of combined treatment was less pronounced than the effect of VD3 or mechanical strain alone. Mechanical strain promoted the gene and protein expression of osteoprotegerin (OPG) and increased the ratio of OPG/RANKL. However, the combination of VD3 and mechanical strain led to a decrease in ratio of OPG/RANKL. Conclusions Mechanical strain might be effective in inducing osteogenic differentiation and increasing bone formation. A joint stimulation with VD3 and strain can decrease proliferation and osteogenic differentiation and increase RANKL expression, which might affect bone remodeling. This study supplies some new data, which might be important in theoretical and clinical research of osteoporosis (OP) and other related bone diseases.
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As one of the important response gene to complement,response gene to complement-32 (RGC-32) simultaneously involves in many other biological functions.Recent studies have shown that RGC-32 was one of the critical regulatory factors at the G2/M check point in the cell cycles and involved in the cell cycle regulation.The expression products of RGC-32 gene play the key roles in cell proliferation,differentiation,inflammation,tumor metastasis and other processes.However,it has not been clarified in its biological mechanisms of regulation in cell cycle.This article takes a brief review about RGC-32 on its gene structure,biological functions,regulation of cell cycle,and the relationship of cell cycle regulation which involves in RGC-32.
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The purpose of the study was to investigate the impact of rat cytomegalovirus(RCMV) infection on the development of the nervous system in rat embryos,and to evaluate the involvement of Wnt signaling pathway key molecules and the downstream gene neurogenin 1(Ngn1) In RCMV infected neural stem cells(NSCs).Infection and control groups were established,each containing 20 pregnant Wistar rats.Rats in the infection group were inoculated with RCMV by intraperitoneal injection on the first day of pregnancy.Rat E20 embryos were taken to evaluate the teratogenic rate.NSCs were isolated from E13 embryos,and maintained in vitro.We found:1) Poor fetal development was found in the infection group with low survival and high malformation rates.2) The proliferation and differentiation of NSCs were affected.In the infection group,NSCs proliferated more slowly and had a lower neurosphere formation rate than the control.The differentiation ratio from NSCs to neurons and glial cells was significantly different from that of the control,showed by immunofluorescence staining.3) Ngn1 mRNA expression and the nuclear β-catenin protein level were significantly lower than the control on day 2 when NSCs differentiated.4) The Morris water maze test was performed on 4-week pups,and the infected rats were found worse in learning and memory ability.In a summary,RCMV infection caused abnormalities in the rat embryonic nervous system,significantly inhibited NSC proliferation and differentiation,and inhibited the expression of key molecules in the Wnt/β-catenin signaling pathway so as to affect NSCs differentiation.This may be an important mechanism by which RCMV causes embryonic nervous system abnormalities.
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Response gene to complement-32(RGC-32)as one of important response genes to complement,involves in many biological functions. Recent studies found that many factors could induce the expression of RGC-32 such as membrane attack complex,cortical hormones,growth factors, luteinizing hormone and transforming growth factor-β and so on. The expression products of RGC-32 play a key role in cell proliferation, differentiation, inflammation, tumor metastasis and other processes, thus to participate in the occurrence of many diseases' development processes. However, it has not be clarified in its exact cell biological function, subcellular localization, gene regulation mechanism. This article will make a brief review on gene structure, functions and roles in cell biology as well as the relationship between human diseases for RGC-32.
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Objective To determine the proliferative activity of HL60 cells by chondroitin sulfate (ChS). MethodsThe proliferative activity was measured using cell counting after incubation with ChS of different concentrations. The acid phosphatase activity of each group was examined with ELISA to observe the differentiation of HL-60 cell. Results The concentration of ChS(
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Objective To investigate the regulatory effect of fluid shear stress(FSS) on the proliferation and differentiation of osteoblasts,as well as the expression of apoptosis-inducing factor,SIVA-1.Methods The third-passage osteoblasts were divided into five experiment groups and one control group.In the experiment groups,1.2 Pa FSS were given to the osteoblasts for 0.25,0.5,1,2,and 4 hours respectively,while the control group received no FSS.Afterwards,the cells were harvested to measure MTT value and ALP activity;mRNA level of SIVA-1 were determined by RT-PCR.Results MTT revealed that the cells proliferation markedly increased in the 0.25 h and 0.5 h experiment groups with advanced cell growth curve;whereas significantly inhibited in 1,2,and 4 h groups.The FSS also increased the ALP activity at 0.25 and 0.5 hour,especially in the 0.5 h group(2.4320?0.205 S unit/100ml,158% of the control;P
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Objective To observe the effect of naringin on proliferation and differentiation of 3T3-L1 preadipocyte,and to investigate its possible mechanism.Methods The cultured 3T3-L1 cells were pretreated with naringin,and then MTT method was used to detect the proliferation of the cells,oil red O staining method and spectrophotography were applied to analyze the degree of differentiation.The expression of PPAR? 2 and C/EBP? mRNA associated with adipocytes differentiation were detected by reverse transcription PCR(RT-PCR).Results Naringin suppressed the proliferation and differentiation of 3T3-L1 preadipocytes and down-regulated the expression of PPAR? 2 and C/EBP? mRNA.Conclusion Naringin can inhibit the proliferation of 3T3-L1,and can suppress their differentiation through down-regulating the expression of PPAR? 2 and C/EBP? mRNA.
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Objective To approach the effect of acrylonitrile (ACN) on the proliferation and differentiation of the lung fibroblasts of mice. Methods The purification and primary culture of the lung fibroblasts(LFb)of new born mice were conducted. The cells were treated with ACN added in the medium at the varying doses of 0.01, 0.5, 1.0, 10.0, 50.0, 100.0 and 200.0 ?g/ml. The cytomorphological methods were used, the protein, malonaldehyde (MDA) and the activity of superoxide dismutase (SOD) were determined. Results No inhibitted effect on the lung fibroblasts was observed at the doses of 0.01-10.0 ?g/ml. At the doses of 50.0-200.0 ?g/ml, acrylonitrile could inhibit the differentiation and proliferation of the cells, the volume of cells became small,the rate of cell clusters decreased, at the doses of 10.0-200.0 ?g/ml, the content of protein and the activity of SOD decreased, MDA content increased. Conclusion Acrylonitrile can inhibit the proliferation and differentiation of the lung fibroblasts in mice, protein synthesis inhibition and lipid peroxidation are considered as the related factors.
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A low calcium medium developed for epidermal keratinocytes were prepared according to the MCDB 153 modified formula and used in human epidermal keratinocyte culture compared with DMEM culture system. The observation by contrast microscopy and electron microscopy showed that in the low calcium medium keratinocytes grew as a monolayer of high proliferation and had many characteristics of basal cells, with a more rounded shape and large intercellular spaces. Increasing the calcium ion concentration in the medium or changing the other culture conditions the cells in these cultures could be induced stratification and terminal differentiation. The results suggest that the growth, proliferation and differentiation of cultured human epidermal keratinocytes can be controlled and regulated someway.