1.Smooth muscle progenitor cells and cardiovascular disease
Chinese Journal of Tissue Engineering Research 2007;0(32):-
It is possible for bone marrow cells,peripheral blood cells,myocardial cells,muscle cells and embryonic stem cells differentiating into smooth muscle progenitor cells(SPCs),which can express special marks of myosin heavy chain.SPCs remained quiescent,until mobilized in response to injury or disease.With various growth factors,cytokines,SPCs are mobilized following physiological stress and subsequently home to site of vascular damage,where they contribute to the remodeling process.Recently,various studies have found that SPCs from different sources contributed to the remodeling process of vessels and development in atherosclerosis.It is generally accepted that SPCs contribute to pathological changes,leads to restenosis.But several recent papers reported findings that injection of SPCs increased collagen and SMC content and reduced macrophage content,consistent with a more stable plaque.SPCs may secrete cytokines and growth factors both locally and systemically that enhance plaque stability.It remains to be determined whether smooth progenitor cells,which are less well studied than their endothelial counterparts,can likewise be manipulated to achieve therapeutic genes benefit.This review has elucidated the origin and distribution of SPCs,mobilization and recruitment,and their roles in cardiac vascular diseases.
2.Transforming growth factor-beta 1 at different concentrations induces the differentiation of bone marrow derived mesenchymal stem cell into smooth muscle-like cell
Yi CHEN ; Wenwei CAI ; Jing SHENG
Chinese Journal of Tissue Engineering Research 2009;13(45):8833-8837
BACKGROUND:Bone marrow derived mesenchymal stem cells (BMSCs) will be homing to the lesions after balloon injury in the inflammatory reaction process.However,the molecular mechanism of transforming growth factor-β1 (TGF-β1) to promote BMSC into smooth muscle-like cell remains unclear.OBJECTIVE:To investigate the BMSC differentiation rate under different TGF-β1 levels after acute vascular injury.DESIGN,TIME AND SETTING:A randomized controlled animal experiment and in vitro induced cell observation were performed at the SPF Laboratory Animal Center and Laboratory of Tissue Engineering,Ninth People's Hospital of Shanghai Jiao Tong University Medical School between January 2008 and May 2009.MATERIALS:A total of 24 male SD rats were randomly divided into normal control and model groups with 12 animals in each group.In addition,6 4-weak-old male SD rats were used to prepare BMSCs.METHODS:Model of acute carotid artery injury was established in model group.Serum of normal control group and model group after 1,3,7 days of injury to detect TGF-β1 level after the vascular injury by ABC-ELISA.The BMSCs after one passage were cultured at a density of (1.0-3.0)×10~5 cells/100 mm in culture dish,and divided into two groups:in routine culture group,cells were cultured in high-glucose DMEM containing 20% fetal bovine serum;in TGF-β1 group,cells were induced by different concentrations of TGF-β1 (1,3,5 and 10 μg/L) based on routine culture for 1 week.MAIN OUTCOME MEASURES:Serum TGF-β1 level after the vascular injury;,cell morphological changes by inverted phase contrast microscopy;smooth muscle α-actin expression.RESULTS:The normal serum TGF-β1 level in rat was low,but increased rapidly after 1 day of injury,reached its peak at 3 days,and declined gradually but did not return to the basic level till day 7.After 1 week of induction,BMSCs were confluent,forming peak valley appearance.Immunocytochemistry showed that compared with routine culture,the rate differentiation of smooth muscle-like cell was significantly increased in cells stimulated by TGF-β1,especially 5 and 10 μg/L TGF-β1 (P < 0.01).Real-time quantitative PCR results were similar to immunocytochemistry.CONCLUSION:Serum TGF-β1 level increased after acute vascular injury and peaked at day 3.In vitro,a similar concentration of TGF-β1 could induce cultured BMSC to differentiate into smooth muscle-like cells.
3.Research progress of small molecule fluorescent probes for ferrous ion and heme
Chen CHEN ; Yi-xin CHEN ; Chong-jing ZHANG
Acta Pharmaceutica Sinica 2023;58(8):2250-2259
Small molecule fluorescent probes have gained widespread attention for their advantages of high selectivity, sensitivity, and easy to operate, and have played a critical role in the detection of various species. They have also demonstrated great potential in the field of biomedical research. Iron, as the most abundant transition metal in the human body, plays a vital role in many physiological functions. Due to the influence of the reductive microenvironment of cell, ferrous ion (Fe2+) is the main component of labile iron in living cells. Heme, consisting of Fe2+ and protoporphyrin IX, is one of the main signaling molecules that wrap biological iron in the human body, and also participates in many physiological and pathological processes. Therefore, the development of small molecule fluorescent probes for detecting Fe2+ and heme as effective monitoring tools will help to further understand their pathological and physiological functions, with potential applications in other fields. This review summarizes the research progress of small molecule fluorescent probes for Fe2+ and heme detection in recent years, and provides insights into future directions for their development.
4.Effects of artesunate on cell proliferation and apoptosis of human Tenon's capsule fibroblasts
Jing CHEN ; Yi CHEN ; Xiulan ZOU ; Yuping ZOU
Recent Advances in Ophthalmology 2017;37(6):523-526
Objective To investigate the effects of artesunate (Art) on cell proliferation and apoptosis of human Tenon's capsule fibroblasts (HTFs),and discuss the countermeasures of bleb scarfing in glaucoma.Methods In vitro,HTFs were cultivated and applicated by different concentrations (50 μg · mL-1,100 μg · mL-1,150 μg ·mL-1,200 μg · mL-1) of Art for 48 hours.The effect of Art on cell proliferation was assessed by MTT method.The rate of apoptosis induced by Art was determined by flow cytometry.Western Blot was performed to detect the relative expression levels of Bax and Bcl-2 after Art was treated.Results After treated with Art for 48 hours,compared with blank control group,Art (50 μg · mL-1,100 μg · mL-1,150 μg · mL-1,200 μg · mL-1) group exhibited notable anti-proliferative effect on HTFs with concentration-dependence (all P < 0.05).The results of flow cytometry showed that the apoptosis rates (8.80% ±0.88%,11.60% ±0.56%,16.30% ±1.03%,23.40% ±1.62%) of HTFs were significantly enhanced with the increase of Art concentration (all P < 0.05).The relative expression levels of Bax were obviously high with the increase of Art concentration,while Bcl-2 levels were significantly low with the increase of Art concentration (all P < 0.05).Conclusion Art can inhibit the proliferation and induce cell apoptosis of HTFs possibly by enhancing the expression of Bax and reducing the expression of Bcl2.Art may be a potential drug in preventing fibrous scar formation after glaucoma filtration surgery.
5.Clinical efficacy comparison of moxibustion with different doses for knee osteoarthritis
Yi-Wen WU ; Ming DAI ; Bi-Song CHEN ; Jing CHEN
Journal of Acupuncture and Tuina Science 2020;18(5):390-395
Objective: To compare the efficacy of moxibustion with different doses for knee osteoarthritis (KOA), and explore the correlation between moxibustion dose and clinical efficacy. Methods: Sixty-eight patients with KOA who met the inclusion criteria were randomly divided into a 20-minute moxibustion group and a 40-minute moxibustion group by the random number table method, with 34 cases in each group. Dubi (ST 35), Neixiyan (EX-LE 4) and Heding (EX-LE 2) were used for moxibustion in the two groups. Each treatment lasted 20 min or 40 min for each point in the 20-minute moxibustion group and 40-minute moxibustion group, separately; the treatment was given 3 times a week and lasted for 4 weeks. The visual analog scale (VAS), Western Ontario and McMaster University osteoarthritis index (WOMAC) and traditional Chinese medicine (TCM) symptom scores were evaluated before and after treatment to compare the efficacy between different moxibustion doses for KOA. Results: After treatment, the total effective rate was 87.5% in the 40-minute moxibustion group, versus 70.0% in the 20-minute moxibustion group, and the difference in the total effective rate between the two groups was statistically significant (P<0.05). After treatment, the VAS scores, the total WOMAC scores and the component scores of pain, stiffness and dysfunction, and the TCM symptom scores in both groups all changed significantly when compared with those before treatment (all P<0.05). After treatment, the between-group differences in the VAS score, the total WOMAC score and the component scores of pain and dysfunction, and the TCM symptom score were statistically significant (all P<0.05), while the difference in the stiffness score in WOMAC showed no statistical significance (P>0.05). Conclusion: Either 20-minute moxibustion or 40-minute moxibustion can relieve pain, improve stiffness, dysfunction, and TCM symptoms for KOA; and 40-minute moxibustion is better in relieving pain, improving dysfunction and TCM symptoms.
6.siRNAs silence expression of mdr1 gene and its role in reversing drug-resistance in K562/ADM cells
Liping GAO ; Hulai WEI ; Tao JING ; Yongjie WU ; Jing CHEN ; Jing SUN ; Juan YI ; Huaishun ZHAO
China Oncology 1998;0(01):-
Background and purpose:Drug-resistance is the main obstacle in terms of efficacy of chemotherapy for leukemia, RNA interference(RNAi) strategy possesses the characteristics of specilization, high-efficiency and low-toxicity, and can effectively and specifically inhibit the overexpression of given gene. This study was designed to investigate the effect of small interfering RNA (siRNA) on expression of mdr1 gene and drug-resistance in multidrug-resistant human leukemia K562/ADM cell.Methods:Human multidrug-resistant leukemia cell line K562/ADM over-expressing mdr1 gene was used as the target cells, Two siRNAs (si-mdr1-1 and si-mdr1-2) targeted mdr1 gene were chemically synthesized and transfected into K562/ADM cells. Expression of mdr1 mRNA was determined by RT-PCR, P-glycoprotein (P-gp) expression was measured using flow cytometry (FCM), and the sensitivity of K562/ADM cells to adriamycin was assessed with a MTT colorimetric assay.Results:Two siRNAs (si-mdr1-1 and si-mdr1-2) specially designed in this study could markedly down-regulate the expression of mdr1 mRNA and its product P-gp in K562/ADM cells. After cells transfected with si-mdr1-1 or si-mdr1-2 for 24h and 48h, the inhibition of mdr1 mRNA expression in the cells for si-mdr1-1 was 55.5% and 22.5%; and for si-mdr1-2, 16.0% and 57.6%, respectively. Treated with siRNA for 72h, the expression intensity of P-gp in the two transfected cell lines decreased 74% and 85%, respectively. Both si-mdr1-1 and si-mdr1-2 significantly enhanced the sensitivity of K562/ADM cells to adriamycin and reversed their drug-resistance, the reversal efficiency was 2.52-folds and 1.96-folds, respectively.Conclusions:The siRNA could effectively and specifically silence the expression of mdr1 gene and overcome the drug-resistance mediated by P-gp in K562/ADM cells.
7.Expression of the transforming growth factor beta-induced gene in human corneal tissue and cell in vitro
Jing-yi, NIU ; Jing, LIU ; Lian, LIU ; Yi-yang, L(U) ; Jian-su, CHEN ; Jin-tang, XU ; Jing-xiang, ZHONG
Chinese Journal of Experimental Ophthalmology 2012;30(1):29-32
Background Clinical studies indicated that the pathogenesis of most corneal dystrophy is associated with the mutation of the transforming growth factor beta-induced (TGFBI) gene.However,the molecular mechanism of mutated TGFBI gene in corneal dystrophy is unclear. Objective The present study was to investigate the expression of the TGFBI gene in human corneal tissue and cells in vitro.MethodsHuman corneal epithelial cells and keratocytes were cultured and passaged,and donor corneal tissue was obtained for the section preparation.RT-PCR was used to detect the expression of TGFBI mRNA in human corneal tissue and cells.Immunofluorescence was used to test the expression of the TGFBI protein in the human corneal tissue,and immunohistochemistry was used to test the expression of the TGFBI protein in human corneal epithelial cells and corneal stromal cells.ResultsRT-PCR analysis showed that TGFBI mRNA could be detected as a 1274 bp band in human corneal tissue and corneal stromal cells,but no TGFBI mRNA was observed in corneal epithelial cells.Immunofluorescence assay revealed that corneal stromal cells were positive ly expressed for the TGFBI protein,but the corneal epithelial cells did not express the TGFBI protein.Immunohistochemistry indicated that the expression of TGFBI was detected the red fluoressence in the cytoplasm of corneal stromal cells;however,no positive response was found in corneal epithelial cells.ConclusionsThe expression of the TGFBI gene occurs in human corneal stromal cells but not in the corneal epithelial cells.This result might be of helpful for studying the function and role of TGFBI gene in pathogenesis of corneal dystrophy.
8.Mechanism about therapeutic effect of meisoindigo on psoriasis via down-regulation of the TLR4-TAK-NF-kappaB pathways.
Haijing ZHANG ; Yi ZHANG ; Jing JIN ; Wanqi ZHOU ; Xiaoguang CHEN
Acta Pharmaceutica Sinica 2013;48(4):503-7
Meisoindigo is an indigo natural derivative commonly used in anti-cancer therapy. In the clinical application, it was also found to have good therapeutic effect on psoriasis. In order to further understand its mechanism of action, human normal keratinocyte cell line HaCaT and RAW 264.7 were used to identify if meisoindigo could affect the inflammatory factors such as NO and other important cytokines which were highly involved in psoriasis. Our results indicated that meisoindigo decreased the production of NO in LPS-stimulated RAW 264.7 cells and reduced the expression of cytokines in LPS-stimulated HaCaT cells. And TLR4-TAK-NF-kappaB was a possible pathway mainly involved in the attenuation of iNOS and pro-inflammatory cytokine production by meisoindigo, which may take part in the therapeutic effect of meisoindigo on psoriasis.
9.Effects of baicalin on CA46 cell proliferation inhibition and apoptosis induction
Yi HUANG ; Jianda HU ; Jing ZHENG ; Tiannan WEI ; Xinji CHEN
Chinese Journal of Pathophysiology 1989;0(05):-
AIM:To investigate the effects of baicalin on proliferation inhibition and apoptosis induction in human Burkitt lymphoma cell line CA46 and to explore its underlying mechanisms.METHODS:CA46 cells were exposed to baicalin at different dosages and its proliferation inhibition was detected by MTT assay.The ability of baicalin to induce CA46 cell apoptosis was examined by Annexin V-FITC/PI double staining analysis,TUNEL labeling method and DNA fragmentation.The mRNA expressions of c-myc and bcl-2 were detected by RT-PCR,and the protein expressions of c-Myc,Bcl-2,caspase-3 precursor(procaspase-3) and poly ADP-ribose polymerase(PARP) were detected by Western blotting.RESULTS:Baicalin remarkably inhibited the CA46 cell proliferation,with an IC50 value of 10 ?mol/L.Apoptosis was remarkably induced by baicalin in a dose-dependent manner,and its earlier and later stages were detected by annexin V-FITC/PI double staining analysis,TUNEL labeling method and DNA fragmentation,respectively.Furthermore,RT-PCR showed that the mRNA expressions of c-myc and bcl-2 in treated CA46 cells decreased in a time-dependent manner.Western blotting showed that the protein expressions of c-Myc,Bcl-2,procaspase-3 and PARP(116 kD) in baicalin treated CA46 cells were down-regulated in a time-dependent manner,while the expression of PARP(85 kD) was up-regulated.CONCLUSION:Baicalin efficiently induces proliferation inhibition and apoptosis in CA46 cells,which may be related with the down-regulation of c-Myc and Bcl-2 expressions,as well as the up-regulation of caspase-3 activity.
10.Effects of morphine on PTEN expression and NF-κB activity in human gastric carcinoma cell line MGC-803
Yi QIN ; Jing CHEN ; Zhiling XIAO ; Yubo XIE ; Qiang XIAO
Chinese Journal of Anesthesiology 2010;30(12):1446-1448
Objective To investigate the effects of morphine on PTEN expression and NF-κB activity in human gastric carcinoma cell line MGC-803.Methods The human gastric cancer cell line MGC-803 was purchased from Cell Biology Research Institute,Chinese Academy of Sciences,and cultured in DMEM liquid culture medium.The cells were randomly divided into 2 groups(n = 6 each): control group and morphine group.The cells was exposed to 0.1 μmol/L morphine in morphine group.The apoptosis was assessed by flow cytometry after being incubated with morphine for 24 h.PTEN expression and NF-κB activity were detected using RT-PCR and Western blot.Results The apoptotic rate was significantly increased,PTEN expression was up-regulated and NF-κB activity was significantly decreased in morphine group compared with control group(P < 0.05).Conclusion Morphine can promote the apoptosis in human gastric cancer cells by up-regulating PTEN expression and decreasing NF-κB activity.