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Total flavones of Abelmoschus manihot L. Medic [TFA] is the major active component isolated from Chinese herb Abelmoschus manihot L. Medic. TFA has shown neuroprotective effect against cerebral ischemia injury in rats and rabbits. However, the effects of TFA on hind-limb ischemia and the underlying mechanisms remain unclear. Therefore, in the present study, we used a rat hind-limb ischemia model to investigate protective effect of TFA against limb ischemia injury. The rat model of hind-limb ischemia was established. Treatment groups received TFA at two different doses [160 and 40mg/kg] daily for 10 days. Sham operated control group and model group received saline. At the end the rats were sacrificed, hindlimb tissues were stained with Haematoxylin-Eosin and Masson's trichrome. RNA and protein were extracted from tissues for PCR and Western blot analysis. The results showed that TFA reduced lower limb ischemic injury, recovered tissue volume and diminished fibrosis and muscle degeneration. Mechanistically, we showed that TFA increased the expression of anti-apoptotic factor such as Bcl-2 and survivin, decreased the expression of pro-apoptotic factor such as Caspase 3, Bax and Bak and inhibited the activation of caspase 3 and 9. In summary, this study proves new evidence that TFA protects hind-limb against ischemia injury by inhibiting apoptosis and could be a promising therapeutic agent for acute lower extremity ischemia
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Objective To investigate the mechanical response of Emerin, a nuclear envelope protein, and its role in apoptosis of vascular smooth muscle cells (VSMCs) during cyclic stretch, and the potential effect of transcriptional factors in this process. Methods Physiological cyclic stretch with the magnitude of 5% and frequency of 1.25 Hz was subjected to VSMCs in vitro by using FX-5000T cyclic stretch loading system. VSMCs cultured under the same conditions but without applying mechanical stretch were used as the static control. The apoptosis of VSMCs was detected by using Cleaved-caspase3 ELISA kit, and the expression of Emerin was revealed by using Western blotting. The effects of Emerin on activities of 345 kinds of transcriptional factors in VSMCs were demonstrated with Protein/DNA array after Emerin specific RNA interference (RNAi) under static condition, and the potential transcriptional factors involved in VSMC apoptosis were analyzed with Ingenurity Pathway Analysis (IPA) software. Furthermore, the binding abilities of Emerin to the motif of 2 kinds of apoptosis-related transcriptional factors were detected with chromatin immunoprecipitation (CHIP) and qPCR. ResultsCompared with the static control, the apoptosis of VSMCs was significantly decreased by 5% cyclic stretch, which suggested a protective effect of physiological cyclic stretch. The expressions of Emerin in VSMCs was remarkably increased with 5% cyclic stretch applied for 6 h, 12 h and 24 h. Specific RNAi under static condition decreased the expressions of Emerin but increased the apoptosis of VSMCs. Emerin siRNA transfection remarkably increased (more than 2 times) the activities of 10 transcriptional factors that participated in cellular apoptosis, i.e. CREB-BP1, p300, p55, MAX, NRF-1, STAT1, STAT3, TEF1, TR and BZP. CHIP-qPCR result revealed that the binding ability of Emerin to specific mofit of STAT1 or STAT3 was significantly repressed with Emerin RNAi. Conclusions Physiological cyclic stretch could increase the expression of Emerin which might modulate the binding of Emerin to motifs of apoptosis-related transcriptional factors such as STAT1 and STAT3, regulate the activities of these factors, and then subsequently repress the VSMC apoptosis. The investigation on mechanobiological mechanisms of VSMC apoptosis induced by cyclic stretch may contribute to further understanding the physiological and pathological mechanisms of vascular homeostasis and vascular remodeling.
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BACKGROUND:Our previous studies have shown that adipose-derived stem cels under vascular endothelial growth factor C (VEGF-C) can be induced to differentiate into lymphatic endothelial cels that are confirmed by lymphatic vascular endothelial hyaluronan receptor-1 staining. However, its optimal induction program is not clear. OBJECTIVE:To investigate the best condition for the differentiation of adipose-derived stem cels into lymphatic endothelial cels under induction of VEGF-C156s. METHODS: Adipose tissues from healthy adults were colected to isolate adipose-derived mesenchymal stem cels using trypsin digestion method. Flow cytometry was employed to detect cel surface markers, andin vitro differentiation capacity was identified by adipogenic and osteogenic induction. Passage 3 cels at good growth state were selected and divided into six groups: cels in control group were cultured in low-glucose DMEM, and those in the rest five groups were treated with 25, 50, 100, 200, 300 μg/L VEGF-C156s, respectively. RESULTS AND CONCLUSION:Adipose-derived stem cels were successfuly obtained by trypsin digestion and purification, and then differentiated into lymphatic endothelial cels under the induction of VEGF-C156s, basic fibroblast growth factor and other growth factors. No cels were positive for lymphatic vascular endothelial hyaluronan receptor-1 in the control group. After 8 days of induction, few cels were positive in the 25 μg/L VEGF-C156s group; a great amount of positive cels were visible in the 50 and 100 μg/L VEGF-C156s groups; 200 and 300 μg/L VEGF-C156s resulted in a large number of deaths in the cels. These findings indicate that it is optimal for adipose-derived stem cels to differentiate into lymphatic endothelial cels under 8-day induction of 50 μg/L VEGF-C156s.
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In order to investigate in greater detail the two methods based on Hertz model for analyzing force-distance curve obtained by atomic force microscopy, we acquired the force-distance curves of Hela and MCF-7 cells by atomic force microscopy (AFM) indentation in this study. After the determination of contact point, Young's modulus in different indentation depth were calculated with two analysis methods of "two point" and "slope fitting". The results showed that the Young's modulus of Hela cell was higher than that of MCF-7 cell,which is in accordance with the F-actin distribution of the two types of cell. We found that the Young's modulus of the cells was decreased with increasing indentation depth and the curve trends by "slope fitting". This indicated that the "slope fitting" method could reduce the error caused by the miscalculation of contact point. The purpose of this study was to provide a guidance for researcher to choose an appropriate method for analyzing AFM indentation force-distance curve.
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Humans , Actins , Elastic Modulus , HeLa Cells , Cell Biology , MCF-7 Cells , Cell Biology , Microscopy, Atomic ForceABSTRACT
Objective To observe the morphologic changes of of vascular buds in vertebral cartilage endplate in age-specific rabbits and also to investigate the correlation between the changes of vascular buds and interverbral disc degeneration. Methods There were 15 New Zealand white rabbits in our study,which include three groups, 2-week-old rabbits, 1-year-old rabbits and 3-year-old rabbits, and each groups had five rabbits. The X-ray radiograph, histology and scanning electron microscope were used to observe the changes of vertebral cartilage endplate. According to Miyamoto standard, the interverbral disc was graded 1-5, and scored 1-5 respectively. Results The changes of micro-vascular structure of vertebral cartilage endplate were observed during aging. Under the scanning electron microscope, the vascular structure degenerated gradually, and disappeared in the end. The blood vessels in the central region of the vertebral cartilage endplate reduced more obviously than those in periphery region. The severe degeneration was found in vertebral endplate, compared with intervertebral disc. The changes of vascular buds in rabbits vertebral cartilage endplate had positive correlation with the vertebral endplate calcification and the interbertebral disc degeneration. Conclusion Changes of vascular buds in vertebral endplate may accelerate intervertebral disc degeneration.
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To investigate effects of collagen matrices by covalent incorporation of heparin and loading with huangqi injection (HI) on anagenetic capillaries, we established the chick chorioallantois model. The collagen matrices by covalent incorporation of heparin and loading with HI were placed and then the eggs were continuously incubated for 3 days. The number of capillaries in the vicinity of samples, the hemoglobin content inside the samples, the dry weight and the macroscopic observation were evaluated. We found the heparinized matrices had comparable angiogenic effects. The number of capillaries, the hemoglobin content, the expression of CD34 increased remarkably (P < 0.01). So we concluded that HI might be considered as an alternative or addition agent to promote the acidification of capillaries.
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Animals , Chick Embryo , Humans , Blood Vessels , Physiology , Capillaries , Metabolism , Collagen , Chemistry , Pharmacology , Drugs, Chinese Herbal , Pharmacology , Heparin , Chemistry , Pharmacology , InjectionsABSTRACT
BACKGROUND: The repair of periodontal tissue is dependent on the number and proliferation and differentiation abilities of periodontal ligament (PDL) cells. PDL cells have the potentiality of multi-directional differentiation such as cementoblast,osteoblast and fibroblast to fonn cement, alveolar bone and periodontal ligament and finally achieve periodontal tissue regeneration.OBJECTIVE:To observe the effects of Shuanghuangbu extract on the proliferation and differentiation of PDL cells.DESIGN:Observation trail.SETTING:Central Laboratory of Wuxi Third People's Hospital. MATERIALS: Periodontal tissue was provided voluntarily by the healthy young patients with deformity correction, and golden thread,skullcap,and rhizoma drynariae by the National Institute for the Control of Pharmaceutical and Biological Products.METHODS: The experiment was conducted in the Central Laboratory of Wuxi Third People's Hospital from July to October 2003.The crushed golden thread.skullcap,and rhizoma drynariae were mixed with distilled water at ratio of 1:10 (m:V),and refluxed in boiling water for 5 hours.The extract was collected,and after colation,the residue was refluxed in boiling water for another 3 hours. Both extract was blended, rotary evaporated and condensed, finally the liquid extract of 3 kg/L was obtained.There were 8 groups in the study including golden thread group, skullcap group,rhizoma drynariae group,golden thread plus skullcap group,golden thread plus rhizoma drynariae group, skullcap plus rhizoma drynariae group, Shuanghuangbu group and control group. PDL cells were cultured/n vitro assisted with Shuanghuangbu.The proliferation of cells was detected with MTT method and the ratio of collagen content in total protein was evaluated with hydroxyproline (HP).MAIN OUTCOME MEASURES:A value of proliferated PDL cells and the proportion of collagen protein in total protein.RESULTS:①Proliferation of PDL cells:Except golden thread group,all Chinese medicine promoted the proliferation of PDL cells significantly compared with control group (P＜0.05). The A value of Shuanghuangbu group was significantly increased.A value was increased with time and reached the peak on day 5.There were significant differences among each group at different time (P＜0.05).②Ratio of collagen content in total protein:Except golden thread group,the percentage was significantly increased by other Chinese medicines compared with control group (P＜0.05), especially Shuanghuangbu. The percentage was increased with time and reached the peak on day 5. There were significant differences among each group at different time (P＜0.05). CONCLUSION:The findings suggest that as a traditional Chinese herb,Shuanghuangbu can significantly stimulate the proliferation and differentiation of PDL cells.and increase the proportion of collagen content in total protein.It may act as an ideal Chinese medicine helper factor for the regeneration of PDL cells.
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0.05) between both groups on re-infarction rate, PCI operation, in-hospital rate of heart diseases and post-operation stroke;while there’s statistical meaning(P
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OBJECTIVE: To analyze the uncertainty involved in HPLC method for the determination Baicalin in Qingkai-ling injection. METHODS: Through establishing HPLC determination mathematical model, the factors affecting the uncertainty were deduced. Each component of uncertainty was evaluated and calculated, from which the combined uncertainty and finally the extended uncertainty and confidence level were obtained. RESULTS: The combined uncertainty was 5.45?10-3, extended uncertainty was 0.01, and the determined content of Baicalin was (4.02?0.01) mg?mL-1. CONCLUSION: The established analytical method is applicable to the uncertainty analysis of HPLC method in the determination of drugs.
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Objective To evaluate the protective effect of estrogen on survival of retinal ganglion cells (RGCs) after transient retinal ischemia-reperfusion (RIR) in rats. Methods RIR was induced in 60 ovariectomized adult rats (OVX) by increasing intraocular pressure via an intracameral catheter. All of the rats were divided into two groups randomly: in experimental group, the rats underwent a subcutaneous injection with 17?-estrodiol(100 ?g/kg) 2 hours before retinal ischemia; and in the control group, saline water was injected correspondingly. The number of RGCs and the thickness of the inner retinal layers were mesured by HE staining method before and 12, 24, 48, and 72 hours after reperfusion. TdT-mediated biotin-dUTP nick end labelling (TUNEL) staining technique was used to examine the apoptosis of RGCs. Results Twenty-four and 48 hours after reperfusion, the number of apoptotic cells in experimental group was obvious lower than that in the control group( P
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Objective: Effect of Zhengjian Granule(ZJ)(Radix Berberidis, Ramulus Euonymi, Radix Scorphulariae Rhizoma Alismatis) on blood pressure(BP) and insulin resistance(IR) in spontaneously hypertensive rats(SHR) was observed. Methods: Twenty four SHR aged 13 weeks were divided into three groups treated with large and small dosage of ZJ and nothing as compared with age matched six Wister Kyoto rats(WKY). BP, glucose(G) and insulin(I) of fasting, G/I and blood lipid were measured before and after treated 8 weeks and compared among groups. Results: While G of fasting had no difference in respective SHR groups compared with WKY, insulin of fasting was markedly higher and G/I were notably lower than WKY( P
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Incised wounds were made in rabbits, and some of the wounds were dressed with chitosan, and others with gauze. After bleeding stopped, hemoglobin in dressings was measured to determine the hemostaic effect. Wounds were redressed, and healing process was studied histologically. The results showed that less Hb was detected in chitosandressing than in gauze. On day 9 after wounding, the content of granulation tissue and collagen formation in the chitosan dressed wounds was more than that in Vaseline gauze dressed group. It suggests that chitosan dressing is effective in hemostasis for incised wounds and may be helpful in wound healing process.