Effect of Lichong decoction on expression of Bcl-2 and Bcl-2-associated X protein mRNAs in hysteromyoma model rat

OBJECTIVE:To study on effects of Lichong decoction on expression of apoptosis-controlling genes,Bcl-2 and Bcl-2-associated X protein(Bax) mRNAs in hysteromyoma tissue of the hysteromyoma model rat.METHODS:Fifty Wistar female rats were randomly divided into a normal group,a model group,a Lichong decoction group,a Guizifuling capsule group and a Mifepristone group.The hysteromyoma rat model was established by intraperitoneal injection of exogenous estrin and progestogens.Pathological examination of uterine tissue,uterine coefficient and uterine transverse diameter were made under optic microscope and expressions of Bcl-2 and Bax mRNAs in uterine tissue in the groups were detected with real-time fluorescent quantitative polymerase chain reaction(PCR) technique.RESULTS:After treatment,under microscope it was found that in the Lichong decoction group myometrium thinned,muscle fiber slightly overgrowth or long and thin,regular arrangement,inserting phenomenon of inner circular muscle and external longitudinal muscle was occasionally or not seen in the Lichong decoction group.The uterine coefficient and the uterine transverse diameter significantly decreased(P<0.01),and Bcl-2 mRNA expression significantly decreased(P<0.01) and Bax mRNA expression significantly increased in hysteromyoma tissue(P<0.01) in the Lichong decoction group as compared with the model group.CONCLUSION:Therapeutic effects of Lichong decoction on hysteromyoma is related with decrease of Bcl-2 mRNA expression and increase of Bax mRNA expression.

Baicalin extracted from Huangqin(Radix Scutellariae Baicalensis) induces apoptosis in gastric cancer cells by regulating B cell lymphoma(Bcl-2)/Bcl-2-associated X protein and activating caspase-3 and caspase-9

OBJECTIVE:To evaluate the effects of baicalin in human gastric cancer cells, including apoptosis-inducing effects, and to investigate its underlying mechanisms of action.METHODS:Cell proliferation and apoptosis assays were performed to investigate the anti-proliferation effects of baicalin in human gastric cancer BGC-823 and MGC-803 cells.Real time-quantitative polymerase chain reaction and Western blotting analysis were performed to elucidate the molecular mechanisms underlying the anti-tumor properties of baicalin.RESULTS:In BGC-823 and MGC-803 gastric cancer cells treated with 80, 120, and 160 μmol/L baicalin for 48 h, a 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide assay showed that baicalin significantly inhibited cell proliferation in a dose-dependent manner, while flow cytometric analysis demonstrated that baicalin could induce apoptosis, also in a dose-dependent manner.Moreover, baicalin up-regulated the expression of caspase-3, caspase-9, and B cell lymphoma(Bcl-2)-associated X protein and down-regulated the expression of Bcl-2 at both the m RNA and protein level.CONCLUSION:Baicalin has potential as a therapeutic agent for gastric cancer by inducing apoptosis in cancer cells.

JTE-522-induced apoptosis in human gastric adenocarinoma cell line AGS cells by caspase activation accompanying cytochrome C release,membrane translocation of Bax and loss of mitochondrial membrane potential

AIM: To investigate the role of the mitochondrial pathway in JTE-522-induced apoptosis and to investigate the relationship between cytochrome C release, caspase activity and loss of mitochondrial membrane potential (Deltapsim). METHODS: Cell culture, cell counting, ELISA assay, TUNEL, flow cytometry, Western blot and fluorometric assay were employed to investigate the effect of JTE-522 on cell proliferation and apoptosis in AGS cells and related molecular mechanism. RESULTS: JTE-522 inhibited the growth of AGS cells and induced the apoptosis. Caspases 8 and 9 were activated during apoptosis as judged by the appearance of cleavage products from procaspase and the caspase activities to cleave specific fluorogenic substrates. To elucidate whether the activation of caspases 8 and 9 was required for the apoptosis induction, we examined the effect of caspase-specific inhibitors on apoptosis. The results showed that caspase inhibitors significantly inhibited the apoptosis induced by JTE-522. In addition, the membrane translocation of Bax and cytosolic release of cytochrome C accompanying with the decrease of the uptake of Rhodamin 123, were detected at an early stage of apoptosis. Furthermore, Bax translocation, cytochrome C release, and caspase 9 activation were blocked by Z-VAD.fmk and Z-IETD-CHO. CONCLUSION: The present data indicate a crucial association between activation of caspases 8, 9, cytochrome C release, membrane translocation of Bax, loss of Deltapsim and JTE-522-induced apoptosis in AGS cells.

Effect of Icariin on apoptosis and expression of Fas,Fas ligand,B cell lymphoma,and Bcl-2-associated X protein in CD4＋ T lymphocytes from patients with ankylosing spondylitis

OBJECTIVE:To investigate the effects of icariin on apoptosis and the expression of Fas, Fas ligand(Fas L), B cell lymphoma(Bcl-2), and Bcl-2-associated X protein(Bax) in CD4+ T lymphocytes from patients with ankylosing spondylitis.METHODS:Primary cultures of peripheral blood CD4+ T lymphocytes were established and treated with icariin at high, medium, and low doses(0.5,0.25, and 0.125 mg/mL).Sulfasalazine treated and helthy cells were used as controls.Apoptosis of treated cells was determined by flow cytometry.Reverse transcription polymerase chain reaction and enzyme-linked immunosorbent assays were used to determine the effects of icariin on the expression of Fas, Fas L, Bcl-2, and Bax.The activity of caspase 8 and caspase 3 was determined by a colorimetric assay.RESULTS:The m RNA and protein expression of Fas,and activity of caspase 8 and caspase 3 in CD4+ T lymphocytes were increased by icariin(P < 0.05).Conversely, the m RNA and protein expression of Bcl-2 was decreased(P < 0.05).The expression of Fas L and Bax were not significantly different between groups.The proapoptotic effects of icariin were dose-dependent.CONCLUSION:Icariin induces the apoptosis of CD4 + T cells from patients with AS comparing to normal control.Therefore, the induction of apoptosis may be the likely mechanism of action of icariin's antirheumatics activities.

Paclitaxel induces apoptosis in human gastric carcinoma cells

AIM;To investigate the apoptosis in gastric cancer cells induced by paclitaxel,and the relation between this apoptosis and expression of Bcl-2 and Bax. METHODS:In in vitro experiments,MTT assay was used to determine the cell growth inhibitory rate.Transmission electron microscope and TUNEL staining method were used to quantitatively and qualitively detect the apoptosis status of gastric cancer cell line SGC-7901 before and after the paclitaxel treatment.Immunohistochemical staining was used to detect the expression of apoptosis-regulated gene Bcl-2 and Bax. RESULTS:Paclitaxel inhibited the growth of gastric cancer cell line SGC-7901 in a dose-and time-dependent manner. Paclitaxel induced SGC-7901 cells to undergo apoptosis with typically apoptotic characteristics,including morphological changes of chromatin condensation,chromatin crescent formation,nucleus fragmentation and apoptotic body formation.Paclitaxel could reduce the expression of apoptosis-regulated gene Bcl-2,and improve the expression of apoptosis-regulated gene Bax. CONCLUSION:Paclitaxel is able to induce the apoptosis in gastric cancer.This apoptosis may be mediated by down- expression of apoptosis-regulated gene Bcl-2 and up- expression of apoptosis-regulated gene Bax.

Effect of quercetin on the expression of Bcl-2/Bax apoptotic proteins in endometrial cells of lipopolysaccharide-induced-abortion mice

OBJECTIVE: To explore the effect of quercetin on the expressions of Bcl-2/Bax apoptotic proteins in endometrial cells in mice with abortion induced by lipopolysaccharide.METHODS: For in vivo experiment, twenty five Kunming mice were randomly divided into five groups at day 4 of pregnancy, with 5 mice per group. The mice were treated with lipopolysaccharide(LPS)through tail vein intravenous injection at day 4 of pregnancy, followed by different concentrations of quercetin by oral gavage consecutively at days 5 to6 of pregnancy. On day 7 of gestation, the mice were sacrificed and the histopathological changes of the uterus tissues were observed. Immunohistochemical staining was applied to the detection of Bcl-2/Bax apoptotic proteins in the endometrial cells. For in vitro experiment, the primary endometrial cells werecultured using a uterus tissue mass culturing method sampled at day 4.5 of pregnancy. The cells were treated with LPS with or without different dosages of quercetin, respectively, for 12 h after 80% confluence. The expression of Bcl-2/Bax apoptotic proteins were detected by western blotting.RESULTS: Both the in vivo and in vitro experiments showed decreased expression of Bcl-2 and enhanced expression of Bax after LPS treatment, leading to a decreased Bcl-2/Bax ratio. The expression of Bcl-2 significantly increased while the expression of Bax was significantly elevated, in the LPS plus quercetin group compared to the LPS only group.CONCLUSION: These results suggest that quercetin has protective effect by partially regulating the expression of Bcl-2/Bax proteins, which in turn inhibits endometrial cell apoptosis and benefits the embryo implantation.

Effect of heparin on apoptosis in human nasopharyngeal carcinoma CNE2 cells

In order to study the mechanism of the effect of heparin on apoptosis in carcinoma cells, the nasopharyngeal carcinoma cell line CNE2 was used to identify the effect of heparin on apoptosis associated with the expression of c-myc, bax, bcl-2 proteins by use of Hoechst 33258 staining, terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL), agarose gel electrophoresis, and flow cytometry, as well as Western blot analysis. The results showed that heparin induced apoptosis of CNE2 cells including the morphologic changes such as reduction in the volume, and the nuclear chromatin condensation, as well as the 'ladder pattern' revealed by agarose gel electrophoresis of DNA in a concentration-dependent manner. The number of TUNEL-positive cells was dramatically increased to 33.6+/-1.2% from 2.8+/-0.3% by treatment with heparin in different concentrations (10 to approximately 40 kU/L). The apoptotic index was increased to 32.5% from 3.5% by detecting SubG1 peaks on flow cytometry. Western blot analysis showed that levels of bcl-2, bax and c-myc were significantly overexpressed by treatment with the increase of heparin concentrations. These results suggest that heparin induces apoptosis of CNE2 cells, which may be regulated by differential expression of apoptosis-related genes.

Effects of erythropoietin on the expression of tumor necrosis factor-alpha and Bax after facial nerve axotomy in rats

This study sought to evaluate the effect of high-dose erythropoietin （EPO; 5 000 IU/kg） on the expression of tumor necrosis factor-alpha （TNF-α） and Bax in the facial nucleus after facial nerve transection in rats. A total of 42 Wistar rats of both genders were used in this study, and 40 rats were randomly divided into 2 groups： EPO group and model group. The EPO group was treated with EPO once a day for 5 days at a dose of 5 000 IU/kg body weight. The model group was treated with saline of the same amount. At day 3 after EPO （or saline） treatment, the right facial nerves of the 40 rats were transected at the level of the stylomastoid foramen, with the left sides untreated. The remaining 2 rats that did not undergo axotomy served as the control group. The surviving motor neurons in operated rats were counted in coronal paraffin sections of the facial nucleus. The expression of TNF-a and Bax in the facial nucleus was detected by immunohistochemical staining at days 3, 7, 14, 21, and 28 after axotomy. At days 14, 21, and 28 after facial nerve axotomy, a significantly greater proportion of facial motor neurons survived in the EPO group than in the model group. After axotomy, the expression of TNF-a and Bax increased in motor neurons in both the EPO and the model groups. TNF-o expression reached its peak level at day 14 after axotomy, while Bax expression reached its peak level at day 21. TNF-α expression was much lower in the EPO group than in the model group at all time points. No significant difference in Bax expression was found between the EPO and the model groups. These results indicate that high-dose EPO treatment attenuates the increase in TNF-α expression in the facial nucleus and reduces the loss of motor neurons after facial nerve transection in rats. However, high-dose EPO treatment has little effect on Bax expression.

Overexpression of Bcl-2 partly inhibits apoptosis of human cervical cancer SiHa cells induced by arsenic trioxide

OBJECTIVE: To study the biological effect of arsenic trioxide (As2O3) on human cervical cancer SiHa cells and SiHa cells overexpressing bcl-2 gene. METHODS: SiHa cells with overexpression of Bcl-2 (SiHa-Bcl2 cells) were established by transfecting SiHa cells with Bcl-2 expression vector. The sensitivities of SiHa and SiHa-Bcl2 cells to As2O3 were determined using MTT (Thiazolyl blue) reduction and colony forming ability assay, morphological analysis, flow cytometric analysis, DNA agarose gel electrophoresis, in situ cell death detection (TUNEL), Northern blot, RT-PCR and Western blot. RESULTS: As2O3 inhibited the growth of SiHa cells and induced G2/M arrest and apoptosis of the cells. RT-PCR and Western blot analysis revealed that As2O3 induced SiHa cell apoptosis possibly via inhibiting the expression of HPV16 E7 and decreasing the expression of c-myc. However, we found that SiHa-Bcl2 cells partly resisted As2O3 induced apoptosis, which might be related to the prevention of the down-regulation of HPV16 E7 and c-myc gene expression. Nevertheless, As2O3 at a high concentration could still induce apoptosis of SiHa-Bcl2 cells mainly via decreasing Bcl-2 expression and slightly inhibiting viral gene expression. CONCLUSION: As2O3 is an inducer of the apoptosis of human cervical carcinoma cells and the cells overexpressing Bcl-2 can partly resist As2O3 induced apoptosis, but the exact mechanism is unclear.

美洲大蠊提取物对实验性肝纤维化大鼠的保护作用

目的探讨美洲大蠊提取物对肝纤维化大鼠的保护作用。方法SD大鼠分为正常对照组(C组)、模型组(M组)、美洲大蠊提取物1组(APA1组)、美洲大蠊提取物2组(APA2组)及还原型谷胱甘肽组(R组)。除正常对照组外,其余4组以40%四氯化碳-橄榄油溶液制备肝纤维化模型,造模同时给予药物干预,于造模7周(C组、M组、APA1组、R组)、9周(APA2组)分2批处死大鼠,评价各组大鼠转氨酶、白蛋白、透明质酸、层连蛋白、肝细胞形态、纤维组织增生及Bax、Bcl-2表达的情况。结果M组大鼠营养状况差,转氨酶、透明质酸、层连蛋白明显升高,血白蛋白显著低于正常,肝细胞广泛坏死,伴有明显纤维组织增生,并形成肝硬化。APA 1组、APA 2组及R组大鼠转氨酶、透明质酸、层连蛋白明显低于模型组,血清白蛋白高于模型组(P<0.05),肝组织肝小叶形态完整,无明显纤维组织增生,中央静脉区肝组织中Bcl-2/Bax比值高于模型组(P<0.05),而汇管区Bcl-2/Bax比值低于模型组(P<0.05)。APA1组大鼠体质量及白蛋白水平高于R组(P<0.05),转氨酶水平略高于R组,肝细胞变性较R组多见。结论美洲大蠊提取物具有保护肝细胞、抑制肝纤维化的作用,对改善肝纤维化大鼠营养状况及调节细胞凋亡具有一定作用。

糖尿病结肠动力障碍近端结肠平滑肌细胞相关凋亡基因表达的研究

目的:探讨糖尿病(DM)结肠动力障碍近端结肠平滑肌细胞的凋亡变化。方法:雄性SD大鼠36只随机分为正常对照6周组、正常对照10周组、DM6周组和DM10周组,每组9只。检测空腹血糖(FBG)、胃肠推进率、血清胰岛素(FINS),HE染色观察近端结肠平滑肌的变化,应用荧光实时定量PCR(Realtime-qPCR)检测大鼠近端结肠平滑肌细胞的bax、bcl-2及caspase-3的mRNA表达情况。结果:(1)DM组大鼠较同时间点正常对照组大鼠空腹血糖增高,胃肠推进率及血清胰岛素降低,近端结肠平滑肌变薄,bax及caspase-3的mRNA表达增强,bcl-2的mRNA表达降低,差异有统计学意义(P<0.01)。(2)DM10周组大鼠较DM6周组大鼠的空腹血糖增高,胃肠推进率及血清胰岛素降低,近端结肠平滑肌变薄,bax及caspase-3的mRNA表达增强,bcl-2的mRNA表达降低,差异有统计学意义(P<0.05)。结论:DM结肠动力障碍与近端结肠平滑肌细胞的凋亡密切相关,且细胞凋亡随病程发展而加重。

Alanyl-glutamine dipeptide inhibits hepatic ischemiareperfusion injury in rats

AIM： To investigate the protective effect and mechanism of alanyl-glutamine dipeptide （Ala-GIn） against hepatic ischemia-reperfusion injury in rats. METHODS： Rats were divided into group C as normal control Group （/7=16） and group G as alanyl-glutamine pretreatment 07=16）. Rats were intravenously infused with 0.9% saline solution in group C and Ala-GIn -enriched （2% glutamine） 0.9% saline solution in group G via central venous catheter for three days. Then all rats underwent hepatic warm ischemia for 30 min followed by different periods of reperfusion. Changes in biochemical parameters, the content of glutathione （GSH） and the activity of superoxide dismutase （SOD） in liver tissue, Bcl-2 and Bax protein expression and morphological changes of liver tissue were compared between both groups. RESULTS： One hour after reperfusion, the levels of liver enzymes in group G were significantly lower than those in group C （P〈0.05）. Twenty-four hours after reperfusion, the levels of liver enzymes in both groups were markedly recovered and the levels of liver enzyme in group G were also significantly lower than those in group C （P〈0.01）. One and 24 h after reperfusion, GSH content in group G was significantly higher than that in group C （P 〈0.05）. There was no statistical difference in activities of SOD between the two groups. One and 24 h after reperfusion, the positive expression rate of Bcl-2 protein was higher in group G than in group C （P〈0.05） and the positive expression rate of Bax protein was lower in group G than in group C （P〈0.05）. Histological and ultrastructural changes of liver tissue were inhibited in group C compared to group G. CONCLUSION： Our results suggest that Ala-GIn pretreatment provides the rat liver with significant tolerance to warm ischemia-reperfusion injury, which may be mediated partially by enhancing GSH content and regulating the expression of Bcl-2 and Bax proteins in the liver tissue.

Regulatory Effects of Zuogui Pill on Apoptosis of Follicles in Rats Injured by 60Co-γRays Based on PI3K/Akt/m TOR Signaling Pathway

[Objectives]To explore the protective effects of Zuogui Pill on ^(60)Co-γ-ray-induced premature aging of rats based on phosphatidylinositol 3-kinase/protein kinase B/mammalian target of rapamycin(PI3K/Akt/mTOR)signaling pathway.[Methods]Sixty sexually mature female SD rats were irradiated with ^(60)Co-γ-ray(6.0 Gy,LD 40)for 24 h at one time.These rats were randomly divided into model group,Progynova group[0.18(g·kg)/d],Progynova[0.09(g·kg)/d]+Zuogui Pill high dose[23.625(g·kg)/d)]group,Zuogui Pill high dose[23.625(g·kg)/d)]group,Zuogui Pill medium dose[9.45(g·kg)/d)]group and Zuogui Pill low dose[4.725(g·kg)/d]group.The administration(once a day)lasted 21 d.The rat serum[follicle-stimulating hormone(FSH),luteinizing hormone(LH)and estradiol(E_(2))]were detected by Enzyme-linked immunosorbent assay(ELISA).The morphological changes of ovary were observed by hematoxylin-eosin(HE)staining.The apoptosis rate of granulosa cells was detected by terminal deoxynucleotidyl transferase(TdT)-mediated dUTP nick-end labeling(TUNEL).The protein expression of phosphorylated(p)-PI3K,p-Akt,p-mTOR,B-cell lymphoma-2(Bcl-2),and Bcl-2-associated X protein(Bax)in ovarian tissues were detected by Western blot.[Results]Compared with the normal group,the model group showed significant increase in the serum FSH(P<0.01),significant decrease in serum E_(2)(P<0.05),and decrease in the number of early follicles and luteum in the ovary(P<0.01).Besides,the apoptosis rate of granulosa cells increased significantly(P<0.01);the expression of p-PI3K,p-Akt,p-mTOR and Bcl-2 in ovarian tissue decreased significantly,while the expression of Bax increased significantly(P<0.01).Compared with the model group,the number of early follicles in the ovary increased and the apoptosis rate of granulosa cells decreased after intervention in each administration group.In addition,the protein expressions of p-PI3K,p-Akt,p-mTOR and Bcl-2 increased,while the expression of Bax decreased,especially in Progynova+Zuogui Pill high dose group,the differences were statistically significant(P<0.05,P<0.01).[Conclusions]Zuogui Pill may protect the radiation-injured ovary through activating the expression of PI3K/Akt/mTOR protein in ovarian tissue,increasing the amount of Bcl-2 protein and inhibiting the expression of Bax protein.

Synergistic impacts of rifampicin and doxorubicin against thioacetamide-induced hepatocellular carcinoma in rats

Background and aims:Combination therapy is a promising new strategy that has been proposed to increase the efficacy of cancer treatment.We aimed to investigate the anti-cancer activity of rifampicin monotherapy and its combination with doxorubicin against hepatocellular carcinoma(HCC).Materials and methods:The in vitro half maximal inhibitory concentration(IC50)and selectivity index(SI)of the drugs under investigation against HepG2 and human lung fibroblast(WI38)cell lines were determined.For the in vivo experiment,male Sprague-Dawley albino rats were injected with thioacetamide at 200 mg/kg twice a week for 90 days;HCC development was confirmed histopathologically.Following HCC induction,the rats were treated with intraperitoneal doxorubicin,rifampicin,or their combination for 45 or 90 days.After sacrifice,the livers were examined histopathologically.The levels of aminotransferases,albumin,bilirubin,malondialdehyde,superoxide dismutase(SOD),catalase(CAT),total antioxidant capacity(TAC),and nitric oxide were measured by spectrophotometry.Alphafetoprotein,cancer antigen 19-9,tumor necrosis factor-alpha,interleukin-6,Bcl-2-associated X protein,caspase 3,caspase 8,and p53 were estimated using ELISA.Results:In vitro,the combination of doxorubicin and rifampicin showed the highest SI of 3.43.In vivo,among the measured markers,the levels of TAC,CAT,SOD,and p53 decreased(P<0.001)and the rest of the measured marker levels increased(P<0.001)in the HCC-bearing rats;after treatment in all groups,all these changes improved toward normal in a time-dependent manner.The combination of doxorubicin and rifampicin optimized the effects of the two individual drugs and exerted the best antioxidant effects.Conclusions:In general,compared with rifampicin or doxorubicin alone,combination therapy has favorable outcomes.Based on our results,the combination of rifampicin and doxorubicin might be applicable for HCC chemotherapy.

Effect of arsenic trioxide on inhibition of restenosis after rabbit vascular injury and its mechanism

OBJECTIVE: To investigate the effect and mechanism of arsenic trioxide (As(2)O(3)) on the prevention of restenosis after vascular injury. METHODS: Apoptosis induction of As(2)O(3) on cultured rabbit vascular smooth muscle cells (VSMCs) in vitro was observed. Thirty-two New Zealand white rabbits were randomly divided into 2- and 4-wk study groups, and their controls. 10% As(2)O(3) at 2.5 mg x Kg(-1) x d(-1) or 0.9% sodium chloride was intraperitoneally infused for 3 days before left common carotid arteries were denudated with a balloon. After denudation 2- and 4-wk animals were sacrificed for morphometry and immunohistochemical studies on carotid arteries, and for histopathology on liver and kidney. RESULTS: It was shown via cellular morphology and DNA fragments in electrophoresis that promotion of As(2)O(3) on cultured vascular smooth muscle cell apoptosis was dependent upon its concentration and duration. Compared with the control animals, the mean vascular intimal proliferation areas were reduced in 2-wk study animals (P 0.05), while the mean vascular luminal areas were all enlarged in both study groups (all P

Effect of Qiangxin Huoli decoction on rats with adriamycin-induced chronic heart failure

OBJECTIVE: To investigate the effect of Qiangxin Huoli decoction on rats with chronic heart failure(CHF) induced by adriamycin(ADR), and to investigate the underlying mechanism of this effect.METHODS: Ninety-six healthy Wistar rats were divided into six groups: control, CHF model, CHF treated by Shenfu injection, and three CHF groups treated with Qiangxin Huoli decoction at high, medium, and low doses, respectively. Qiangxin Huoli decoction was administered orally to protect the stomach in the three Qiangxin Huoli decoction groups, while the control group and the CHF model group were administered the same volume of 0.9% physiological saline, and the Shenfu group wereadministered the same volume of Shenfu injection. Ten days later, the CHF model was then induced in all groups except the control group by intraperitoneal injection of ADR at gradient dose intervals. The bodyweights were recorded on days 10, 20, 30, and 40. Hemodynamic indices were recorded, including left ventricular systolic pressure(LVSP), left ventricular end-diastolic pressure(LVEDP), maximum increase in left ventricular pressure(+dp/dt_(max)), maximum decrease in left ventricular pressure(-dp/dt_(max)), heart rate(HR), and electrocardiogram using an eight-channel physiological recorder with LabChart software monitoring. The plasma brain natriuretic peptide(BNP) concentration was determined by enzyme-linked immunosorbent adsorption. The expressions of B-cell lymphoma-2(Bcl-2) and Bcl-2-associated X protein(Bax) were detected by immunohistochemical methods.RESULTS: The CHF model group were in poor condition, and the mean bodyweight was significantly decreased compared with the control group. Furthermore, compared with the control group, the CHF groups had significantly decreased LVSP, +dp/dt_(max), and-dp/dt_(max), and significantly increased LVEDP. The CHF groups also showed significant increases in HR, S-T segment elevation, and plasma BNP levels compared with the control group. Compared with the CHF model group, the treatment groups had significantly increased Bax expression(P < 0.05) and significantly decreased Bcl-2 expression(P < 0.01), indicating less apoptosis. The high dose Qiangxin Huoli decoction group and the Shenfu group showed the most significant improvements.CONCLUSION: In the rat model of CHF, Qiangxin Huoli decoction significantly reduces the abnormal hemodynamics, improves cardiac function, reduces plasma BNP concentration, regulates the expression of apoptosis proteins, inhibits the apoptosis of myocardial cells, and plays a protective role.

Inhibitory effect of oridonin on proliferation of RPMI8226 cells and the possible underlying mechanism

OBJECTIVE:To observe the effects of oridonin on proliferation and apoptosis of myeloma RPMI8226cells and to investigate the potential underlying mechanisms.METHODS:RPMI8226 cells were treated with various concentrations of oridonin.Cell proliferation was analyzed using the thiazolyl blue tetrazolium bromide method.Ultramicrostructure was observed by transmission electron microscopy.Annexin-V/PI staining and flow cytometry was performed to determine cell apoptosis.Expression of apoptosis-related proteins was evaluated by western blot analysis.RESULTS:Oridonin suppressed the proliferation of RPMI8226 cells and induced apoptosis in a timeand dose-dependent manner.Transmission electron microscopy confirmed apoptotic morphologyupon treatment with 20μmol/L oridonin and western blot revealed decreased expressions of the apoptosis suppressors survivin,Bcl-2 and pro-caspase-3 proteins,and the increased expression of the apoptosis inducer Bax.CONCLUSION:Our results show that oridonin exhibits an inhibitory effect on the proliferation of RPMI8226 cells and induces apoptosis.This is associated with altering the balance between Bcl-2 and Bax protein expressions and decreased survivin and pro-caspase-3 expressions.

Molecular mechanism of damage and repair of mouse thymus lymphocytes induced by radiation

OBJECTIVE: To investigate the role of apoptosis in radiation-induced mouse thymus lymphocyte damage and repair and provide the basis for understanding the molecular mechanism of radiation-induced lymphocyte damage and repair as well as the prevention and treatment of acute radiation sickness. METHODS: We studied the dynamic changes of apoptosis of mouse thymus lymphocytes and the expression of bax and bcl-2 gene products after 2, 4, 6 and 8 Gy of whole body gamma-irradiation using in situ terminal labeling, DNA electrophoresis and immunohistochemical techniques. RESULTS: At the early stage after irradiation, the percentage of apoptotic lymphocytes increased rapidly in accordance with the increasing of radiation doses, while the counts of the thymus and peripheral lymphocytes decreased sharply, showing an opposite change to lymphocyte apoptosis. After 6 Gy gamma-irradiation, typical morphological characteristics of thymus apoptotic lymphocytes in early, middle and late stages were found by transmission electron microscopy. The thymus lymphocytes displayed characteristic DNA ladders 4 hr and 8 hr after 2-6 Gy gamma-irradiation,using DNA gel electrophoresis techniques. Abnormal expression of bcl-2 and bax gene products were shown in irradiated lymphocytes. CONCLUSIONS: Apoptosis plays an important role in the process of radiation-induced mouse thymus lymphocyte damage and repair. Bcl-2 and Bax proteins may regulate the process of lymphocyte apoptosis.

木犀草素对小鼠T淋巴细胞体外增殖及凋亡的作用

目的：研究木犀草素对小鼠T淋巴细胞体外增殖及凋亡作用的影响。方法：设置刀豆蛋白A（Con A）组和空白组,设置3个木犀草素药物组,终质量浓度分别为2.5,5,10 mg·L-1,细胞毒性活性检测-8（CCK-8）法检测各组细胞增殖情况,末端脱氧核苷酸转移酶介导的d UTP缺口末端标记测定法（TUNEL）和Annexin V-FITC/PI双染法检测各组细胞凋亡情况,蛋白免疫印迹法（Western blot）检测各组细胞凋亡通路蛋白B淋巴细胞瘤-2（B-cell lymphoma-2,Bcl-2）,Bcl-2相关X蛋白（Bcl-2-associated X protein,Bax）表达水平。结果：CCK-8实验结果显示3个木犀草素药物组细胞数明显低于Con A组和空白组（P〈0.01）,差异有统计学意义。TUNEL实验结果显示,Con A组的凋亡细胞数量显著低于3个木犀草素药物组（P〈0.01）,差异有统计学意义。流式细胞仪检测结果显示3个木犀草素药物组的细胞凋亡率显著高于Con A组和空白组（P〈0.01）,细胞凋亡率随着药物浓度增加而增高,呈明显的量-效关系。Western blot检测结果显示,2.5,5 mg·L-1木犀草素药物组中Bcl-2表达量高于对照组,Bax表达量低于对照组;10 mg·L-1药物组中Bcl-2表达量低于对照组,Bax表达量高于对照组。结论：2.5,5,10 mg·L-1木犀草素能够抑制T淋巴细胞增殖,能够诱导T淋巴细胞凋亡,10 mg·L-1木犀草素可能主要通过下调Bcl-2,上调Bax影响线粒体凋亡通路诱导细胞凋亡,2.5,5 mg·L-1木犀草素可能通过其他通路诱导细胞凋亡。