A microarray analysis of angiogenesis modulation effect of Xuefu Zhuyu Decoction on endothelial cells / 中国结合医学杂志

<p><b>OBJECTIVE</b>To study the angiogenesis modulation mechanism of Xuefu Zhuyu Decoction () on the endothelial cell line ECV304.</p><p><b>METHODS</b>ECV304 cells were treated with 2.5% Xuefu Zhuyu Decoction-containing serum (XFZYD-CS) for 24 h, 48 h or 72 h. Thiazolyl blue tetrazolium bromide (MTT), fluorescence activating cell sorter (FACS), migration, adhesion and in vitro tube formation assays were conducted to confirm an angiogenesis effect of XFZYD at 3 time points. An analysis of angiogenesis regulator profiles was performed at 3 times with real-time polymerase chain reaction (RT-PCR) superarray.</p><p><b>RESULTS</b>At 48 h, XFZYD-CS induced ECV304 significantly improved cell viability, number in S phase, migration, adhesion and tube formation. At 24 h and 72 h, only cell migration was elevated. Microarray results showed that the expression of 27 angiogenesis-related genes was changed.</p><p><b>CONCLUSION</b>XFZYD-CS treatment induced angiogenesis on ECV304 cells with significant cellcular changes occurring at 48 h and genetic changes as early as 24 h.</p>

Study on function of VEGF pathway in promoting angiogenesis with Xuefu Zhuyu Tang / 中国中药杂志

<p><b>OBJECTIVE</b>To investigate the function of VEGF pathway in promoting angiogenesis with Xuefu Zhuyu Tang (XFZYT).</p><p><b>METHOD</b>Serum pharmacology technique was adopted to treat endothelial cells ECV304 with XFZYD containing serum and blank serum with concentrations of 1.25%, 2.5% and 5%. Methyl thiazolyl tetrazolium (MTT) assay, boyden chamber migration assay and in vitro vessel formation assay were used to test endothelial cell proliferation, migration and angiogenesis capacities. ELISA, immunohistochemistry and RT-PCR were used to detect secretion and expression of vascular endothelial growth factor (VEGF) and vascular endothelial growth factor receptor-2 (VEGFR2).</p><p><b>RESULT</b>XFZYT-CS with a concentration of 1.25% only affected angiogenesis capacity in vitro. XFZYT-CS with a concentration of 2. 5% promoted cell proliferation, migration and angiogenesis capacities and significantly increased VEGF level and VEGF/VEGFR2 expressions. XFZYT-CS with a concentration of 5% only up-regulated intracellular VEGF expression and thereby improving endothelial cell proliferation, migration and angiogenesis.</p><p><b>CONCLUSION</b>XFZYT can induce endothelial cell proliferation, migration and angiogenesis by up-regulating VEGF-VEGFR2 pathway, which partially proves its angiogenesis effects.</p>

Experimental study on the suppression of sodium nitroprussiate-induced chondrocyte apoptosis by Tougu Xiaotong Capsule (透骨消痛胶囊)-containing serum / 中国结合医学杂志

<p><b>OBJECTIVE</b>To study the mechanism of action of Tougu Xiaotong Capsule (透骨消痛胶囊, TGXTC) ex vivo in suppressing chondrocyte (CD) apoptosis induced by sodium nitroprussiate (SNP).</p><p><b>METHODS</b>Thirty New Zealand rabbits, 2 months old, were randomized by lottery into five groups, six in each: the blank group treated with saline, the positive control group treated with Zhuanggu Guanjie Pill (壮骨关节丸, 70 mg/kg), and the three experimental groups, EGA, EGB, and EGC, treated with low dose (35 mg/kg), moderate dose (70 mg/kg), and high dose (140 mg/kg) of TGXTC, respectively. All treatments were administered via gastrogavage twice a day for 3 days. Arterial blood was collected from the abdominal aorta and drug or drug metabolites-containing serum was prepared. CDs obtained from knee joints of 16 four-week-old New Zealand rabbits were cultured to the third passage and confirmed by toluidine blue staining. SNP of various final concentrations (0, 0.5, 1.0, and 2.0 mmol/L) was used to induce CD apoptosis, and the dosage-effect relationship of SNP in inducing CD apoptosis was determined. Serum samples from the blank, control, and three dosages of TGXTC-treated rabbits were tested in the CD culture in the presence of SNP. Cell apoptosis was determined by Hoechst 33342 staining, viability of CDs was quantified by MTT, CD apoptosis rate was determined by annexin V-FITC/PI staining, levels of p53 and Bcl-2 mRNA expression in CDs were determined with RT-PCR, and contents of caspase-3 and caspase-9 proteins were determined by colorimetry.</p><p><b>RESULTS</b>CD apoptosis was induced by SNP at all concentrations tested and in a dose-dependent manner. The SNP concentration of 1 mmol/L and treatment duration of 24 h appeared to be optimal and were selected for the study. Serum samples from the positive control rabbits and from the two higher doses of TGXTC-treated rabbits showed reduction of SNP-induced CD apoptosis, decrease in p53 mRNA expression, inhibition of catalytic activities of caspase-3 and caspase-9, and increase in Bcl-2 mRNA expression when compared with the serum from the blank group (P<0.05).</p><p><b>CONCLUSION</b>TGXTC-containing sera antagonized SNP-induced CD apoptosis and the molecular basis for the action was associated with up-regulation of Bcl-2, down-regulation of p53 expression, and inhibition of caspase-3 and caspase-9 catalytic activities.</p>

Effects of time factors on acupoint sticking therapy for preventing and treating bronchial asthma / 中国针灸

<p><b>OBJECTIVE</b>To observe the influence of time factors on acupoint sticking therapy for preventing and treating bronchial asthma.</p><p><b>METHODS</b>Seventy-one cases were randomly divided into a dog days group (n= 30), a Sanjiu days group (n=21) and a daily days group (n=20). They were all treated with ginger moxibustion plus acupoint sticking of Chinese medicine at Dazhui (GV 14) and Feishu (BL 13) etc. This treatment was applied once at the beginning of the first dog days, the middle dog days and the last dog days respectively in the dog days group, and once at the beginning of the first nine days, the middle nine days and the last nine days in coldest days of winter respectively in the Sanjiu days group, and once every other 10 days during 30 days except the dog days or the Sanjiu days in the daily days group. Their therapeutic effects and quality of life and changes of serum level of interleukin 13 (IL-13) were observed.</p><p><b>RESULTS</b>The total effective rate of the dog days group was 83.3% (25/30), the Sanjiu days group and the daily days group were 61.9% (13/21) and 65.0% (15/20) respectively, with no significant differences among three groups (all P>0.05). After treatment, there were no significant differences in quality of life and changes of serum level of IL-13 among three groups, but there were significant differences between before and after treatment (P<0.01, P<0.001).</p><p><b>CONCLUSION</b>Acupoint sticking is an effective therapy for bronchial asthma. It can be practiced in the whole year for the result of this study that medicines and acupoints are the leading factors of this therapy and the time factors have no influence on therapeutic effect.</p>

Protection of apoptosis of osteoblast cultured in vitro by Morinda Root Polysaccharide / 中国骨伤

<p><b>OBJECTIVE</b>To explore the protection on apoptosis and the mechanism of promoting the cytoactive of osteoblast by Morinda Root Polysaccharide through the observations of the cultured osteoblast in vitro.</p><p><b>METHODS</b>Prepared blood serum with Morinda Root Polysaccharide and Morinda Root aqueous extract and cultured Osteoblast in vitro with it. The second generation osteoblasts in vitro were separated from the cranium of 24-hours newborn SD rat, which were divided into control group (adding only rat serum during cultivation), induction apoptosis group (adding trans-retinoic acid in control group), Morinda Root aqueous extract group (adding serum prepared by Morinda Root aqueous extract in induction apoptosis group) and Morinda Root Polysaccharide group (adding serum prepared by Morinda Root Polysaccharide in induction apoptosis group). Adopting fluorescence microscope, apoptosis detected by flow cytometry and gene expression of Bcl-2 and Bax detected by RT-PCR, to evaluate the effect of Morinda Root Polysaccharide on the course of osteoblast apoptosis.</p><p><b>RESULTS</b>The apoptotic rate of Morinda Root aqueous extract group and Morinda Root Polysaccharide group were significantly lower than that of induction apoptosis group (P < 0.01). The apoptosis ratio of Morinda Root Polysaccharide group was lower than that of Morinda Root aqueous extract group (P < 0.05). Expression level of Bcl-2 mRNA of apoptosis cell: control group > Morinda Root Polysaccharide group > Morinda Root aqueous extract group > induction apoptosis group (P < 0.01). Expression level of Bax mRNA: induction apoptosis group > Morinda Root aqueous extract group > control group > Morinda Root Polysaccharide group (P < 0.01). Bcl-2/Bax: control group > Morinda Root Polysaccharide group > Morinda Root aqueous extract group > induction apoptosis group (P < 0.01).</p><p><b>CONCLUSION</b>Morinda Root can inhibit the apoptosis of osteoblast induced by trans-retinoic acid in some extent. The above role of Morinda Root Polysaccharide is significant better than that of Morinda Root aqueous extract. It is indicated that Morinda Root Polysaccharide is one of the essential component of inhibiting osteoblast apotosis.</p>

Experimental study on the effects of pharmaco-serum of diabetic rats fed with Chinese herbs qianggubao decoction (Chinese characters: see text) on osteoblast culture in vitro / 中国骨伤

<p><b>OBJECTIVE</b>To investigate the optimum phase and dose of pharmaco-serum of diabetic rats fed with Qianggubao decoction ([Chinese characters: see text]) on the differentiation and mineralization of osteoblast (OB). METHODS (OB) was isolated from the skull of 10 newly born SD rats aged 1 to 2 days by means of Trypsin-collagenase digestion. After the OB was identified, different kinds of pharmaco-serum of diabetic rats fed with inactive Qianggubao decoction ([Chinese characters: see text]) of different phase (rats were fed with medicine three days or five days after last fed with medicine one hour or three hours) and concentration (5%, 10%, 20%) were added to the OB and incubated. After 7 days and 18 days of culture,the effects of the differentiation and mineralization of osteoblast were detected.</p><p><b>RESULTS</b>The secretion of ALP and formation of mineralized nodules of osteoblast in the different doses of pharmaco-serum groups were almost the same as that of normal control group, but were superior to that in the model control group. And the group with concentration of 20% pharmaco-serum was the best in the secretion of ALP and formation of mineralized nodules of osteoblast. As to the phases of pharmaco-serum, the best one on the differentiation and mineralization of osteoblast was the serums from diabetic rat-model fed with Qianggubao decoction ([Chinese characters: see text]) three days or five days, after one hour of last fed with medicine.</p><p><b>CONCLUSION</b>The pharmaco-serum of diabetic rats fed with Qianggubao decoction ([Chinese characters: see text]) can promote the differentiation and mineralization of osteoblast. Allow for time and the cost of experiment,we presume that pharmaco-serum of diabetic rats fed with Qianggubao decoction ([Chinese characters: see text]) three days, after one hour of last fed, with concentration of 20% and not-inactivation is the optimum on the differentiation and mineralization of osteoblast.</p>

Experimental study on the effects of inactivated and un-inactivated pharmaco-serum of diabetic rats fed with Chinese herbs Qianggubao decoction on the proliferation of osteoblast cultured in vitro / 中国骨伤

<p><b>OBJECTIVE</b>To study the effect of inactivated and un-inactivated pharmaco-serum of diabetic rats fed with Chinese herbs Qianggubao decoction on the proliferation of osteoblast cells (OB)cultured in vitro.</p><p><b>METHODS</b>OB was isolated from the skull of newly born SD rats aged 1 to 2 days by means of Trypsin-collagenase digestion and identified by image analysis under inverted microscope, V-G collagen staining, ALP staining, calcification nod staining etc. After the OB was identified, in activated and un-inactivated pharmaco-serum of diabetic rats fed with Qianggubao decoction of ferent phase (rats were fed with medicine 3 days or 5 days after last fed with medicine 1 hour or 3 hours) and concentration (5%, 10%, 20%) were added to the OB and incubated. After determined times, the effects of the proliferation of osteoblasts were detected by MTT analysis.</p><p><b>RESULTS</b>There was significant difference between un-inactivated pharmaco-serum and inactivated pharmaco-serum on the proliferation of osteoblasts, and un-inactivated serum had stronger effects to improve the proliferation of osteoblasts (P < 0.01 or P < 0.05).</p><p><b>CONCLUSION</b>Un-inactivated and inactivation pharmaco-serum of diabetic rats fed with Chinese herbs Qianggubao decoction can influence the proliferation of, and the un-inactivated pharmaco-serum has stronger effects.</p>

Experimental study on effect of Xuefu Zhuyu Decoction on bone marrow hematopoietic stem cells of mice / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To investigate the effect of Xuefu Zhuyu Decoction (XFZYD) on the number, phenotype, cell cycle and colony formation of bone marrow hematopoietic stem cells (HSC) in mice.</p><p><b>METHODS</b>Kunming mice were randomly divided into 4 groups: the control group, the low- (3.25 g/kg), middle- (6.5 g/kg) and high-dose (13.0 g/kg) XFZYD groups. After they were medicated by gastrogavage respectively with saline or corresponding dose of XFZYD for 7 days, their bone marrow HSC were separated and counted. The phenotype Sca and cell cycle of HSC were detected by flow cytometer, and the colony formation was determined with semisolid methyl media culture.</p><p><b>RESULTS</b>No obvious difference in the number of mononuclear cell, suspended cell and colony production was found among all the groups (P > 0.05); while the expression of CD34 and Sca-1 increased in the low-dose XFZYD group, but in the middle-dose XFZYD group increase only showed in Sca-1 expression.</p><p><b>CONCLUSION</b>XFZYD plays a role of removing blood stasis and promoting regeneration through improving hematopoietic function by means of increasing the number and enhancing the function of premature HSC.</p>