Effect of LncRNA p21 regulating Hippo-YAP signaling pathway on formation of abdominal aortic aneurysm in mice and its mechanism / 中国药理学通报

Aim To investigate the effect of long non- coding RNA p21 (LncRNA p21) regulating Hippo- Yes-associated protein (Hippo-YAP) signaling pathway on the formation of abdominal aortic aneurysm (AAA) in mice. Methods C57BL/6 ApoE

Effect of dexmedetomidine on cell apoptosis during lung ischemia-reperfusion injury in a rat model of cardiopulmonary bypass / 中华麻醉学杂志

Objective To evaluate the effect of dexmedetomidine on cell apoptosis during lung is-chemia-reperfusion(I/R)injury in a rat model of cardiopulmonary bypass(CPB).Methods Ninety-six SPF healthy adult male Sprague-Dawley rats,weighing 350-500 g,were divided into 4 groups(n＝24 each)using a random number table method: sham operation group(group S),CPB group(group C),CPB plus left lung I/R group(group IR),and CPB plus left lung I/R plus dexmedetomidine group(group D).The chest was only opened,and the rats underwent no CPB in group S.Only the CPB model was es-tablished in group C.The model of left lung I/R injury was established based on the CPB model in group IR.In group D,the model of CPB plus left pulmonary I/R injury was established,dexmedetomidine was intrave-nously infused in a dose of 3 μg/kg through the tail vein,followed by a continuous infusion of 1.5 μg?kg-1?h-1 until the end of surgery.Eight rats were selected in each group before operation(T0),at 10 min after opening the left hilum(T1),and at the end of operation(T2),the left lung tissues were taken for examination of pathological changes(with a light microscope)which were scored and for determination of cell apoptosis,and immunohistochemistry score(IHS)was assessed.The apoptosis index was calculated.Results Compared with group S,the pathological changes of lung tissues,IHS and apoptosis index were significantly increased at T1,2 in the other three groups(P＜0.05).Compared with group C,the pathologi-cal changes of lung tissues,IHS and apoptosis index were significantly increased at T1,2 in IR and D groups(P＜0.05).Compared with group IR,the pathological changes of lung tissues,IHS and apoptosis index were significantly decreased at T2 in group D(P＜0.05).Conclusion The mechanism by which dexme-detomidine reduces lung I/R injury during CPB is related to inhibiting cell apoptosis in rats.

Norcantharidin enhanced cisplatin sensitivity via YAP pathway in A549 cells / 中国药理学通报

Aim To investigate the effect of cotreat-ment norcantharidin (NCTD) and cisplatin (DDP) on the cisplatin sensitivity and proliferation in A549 cells, and whether YAP molecule involved in this process. Methods A549 cells were used as a model for inves-tigating the function of NCTD and DDP in this study. The expression of caspase-3,Annexin V,YAP,CTGF and Cyr61 were detected using RT-PCR and Western blot assay. The cell growth and proliferation were as-sessed by MTT and CCK-8 assay. Moreover, the tran-scriptional activity of YAP was detected by luciferase reporter gene assays. Results YAP was required for the cell growth and proliferation. NCTD,DDP and co-treatment of NCTD and DDP depressed cell viability, inhibited cell proliferation and promoted the sensitivity of cisplatin in A549 cells. Our results showed that the higher expressed oncogene YAP activated and promoted cell proliferation in lung cancer cells. Moreover, the high concentration of NCTD or DDP significantly re-duced cell proliferation, but cotreatment of NCTD and DDP in low concentration could significantly increase the cisplatin sensitivity via YAP pathway in A549 cells. Conclusions The cotreatment of NCTD and DDP in low concentration significantly reduces the transcriptional activity and protein level of YAP, then inhibits cell proliferation and thus increases the sensi-tivity of DDP in A549 cells.

The effect of mechanical stretching stress on the proliferation, apoptosis of degenerative disc nucleus pulposus cells and expression of extracellular matrix / 中华物理医学与康复杂志

Objective To observe in vitro the effect of mechanical stress at different intensities on the pro-liferation, apoptosis and extracellular matrix of degenerative human nucleus pulposus cells. Methods The cells were isolated and cultured in vitro, and divided into a control group, a low-intensity group, a medium-intensity group and a high-intensity group. The low-, medium- and high-intensity groups were stretched mechanically by 1000 μ, 2000μor 4000μrespectively for 6 hours using a four-point bending system, while the control group was not stressed. Flow cytometry was used to explore any changes in the cell cycle and the proliferation index ( PI) . The expression of proliferating cell nuclear antigen ( PCNA) , B-cell lymphoma-2 ( BCL-2)/Bax, collagen II and aggrecan were meas-ured using real-time quantitative polymerase chain reactions. Results The mechanical stretching significantly influ-enced proliferation, apoptosis and the extracellular matrices compared with the control group. The PI and PCNA ex-pression increased at first and then decreased gradually with the exercise intensity. Compared with the control group, the mRNA expression level of Bcl-2/Bax increased significantly to 1.53 times that of the control group after 1000 μstretching, but to only 0.71 times that of the control group at 2000μand 0.43 times at 4000μ. The gene expression of collagen II increased significantly by 1. 1 times and that of aggrecan by 1. 3 times after 1000 μ stress stimulation compared with the control group ( P≤0.05) . However, the expression of collagen II and aggrecan was inhibited sig-nificantly at 2000 and 4000 μ , with the lowest levels at 4000 μ (P≤0.05). Conclusion Stretching at different intensities has different effects on the proliferation, apoptosis and extracellular matrix of human pulposus cells with degenerate nuclei.

Ginkgolide B inhibits apoptosis in high glucose-stimulated human umbilical vein endothelial cells / 中国药理学通报

Aim Toinvestigatetheeffectofginkgolide B on apoptosis in high glucose-treated endothelial cells.Methods Humanumbilicalveinendothelial cells(HUVECs)were used in the present study.The level of transmigration of HUVECs was analyzed by Tr-answell experiment.Apoptosis was detected by flow cy-tometry.Reactive Oxygen Species (ROS ) was meas-ured by immunofluorescence kit.The protein expres-sionwasanalyzedbyWesternblot.Result Highglu-cose treatment resulted in a reduction in transmigration of HUVECs and ginkgolide B recovered the phenome-non in glucose-treated endothelial cells.The level of ROS generation was increased in high glucose-treated group,whereas ginkgolide B inhibited ROS genera-tion.Immunofluorescence data showed high glucose in-creased apoptosis,whereas ginkgolide B inhibited ap-optosis in high glucose-treated HUVECs.Moreover, the expressions of Bax and caspase-3 were increased and Bcl-2 was reduced in high glucose-treated group. In contrast,ginkgolide B abolished the expressions of Bax and caspase-3 and increased Bcl-2 expression. Moreover,high glucose enhanced the expression and phosphorylation of p53,while ginkgolide B suppressed the expression and phosphorylation of p53 induced by highglucose.Conclusions GinkgolideBcaninhibit apoptosis and improve transmigration function in high glucose-treated HUVECs.Ginkgolide B has protection against high glucose-induced endothelial cell injury.

Effect of plumbagin on invasion and apoptosis of human hepatocellular carcinoma cell line HepG2 / 中国药理学通报

Aim To investigate the effect of plumbagin on invasion and apoptosis of human hepatocellular car-cinoma ( HepG2 ) . Methods HepG2 cells were cul-tured in vitro with different concentrations of plumba-gin, then cell proliferation was observed by MTT as-say; cell invasion was observed by transwell invasion assay; cell apoptosis was detected by flow cytometry, and the protein expression of Bax, Bcl-2 was detected by immunocytochemistry. Results MTT results showed that plumbagin could significantly inhibit cell proliferation compared with the control group, and in a dose-dependent manner ( P ( P < 0. 01 ) . Flow cytometry showed that apoptosis rate was significantly higher in 4 , 8 μmol · L-1 group of plumbagin compared with that of the control group ( P<0. 01 ) . Immunocytochemistry showed that, with the increasing concentration of plumbagin, Bax protein expression increased, Bcl-2 protein expression was de-creased, both in a dose-dependent manner ( P <0. 01 ) . Conclusion Plumbagin can inhibit HepG2 cell proliferation and accelerate apoptosis of HepG2 cells, but also has the ability to inhibit HepG2 cell in-vasion.

Digital holographic mi cor scopyf or detectin g galangin-induced apopt osis of HepG2 cells / 医学研究生学报

Objective Galangin has various antitumor activities and digital holographic microscopy ( DHM ) .This study aimed to investigate the detection of galangin-induced apoptosis of HepG2 cells using DHM. Methods HepG2 cells were cultured in vitro and treated with galangin at 130 μmol/L for 6, 12, and 24 hours respectively to induce apoptosis.Then, MTT assay was used to detect the proliferation of the HepG2 cells, Hoechst 33342 staining and flow cytometry adopted to determine their apoptosis, and DHM employed to visualize their morphological changes. Results Compared with the blank controls, galangin significantly inhibited the proliferation of the HepG2 cells ([7.32 ±2.47]%vs [39.14 ±2.04]%, P<0.01) and obviously induced their nuclear condensa-tion and apoptosis, with a total apoptosis rate of (11.550 ±0.043)%at 24 hours.DHM showed statistically significant differences be-tween the galangin-treated and control groups in the morphological changes of the HepG2 cells at the three time points (P<0.05). Conclusion Digital holographic microscopy can be used to detect the morphological changes of HepG2 cells during galangin-induced apop-tosis.

Role of NADPH oxidase in high glucose-induced injury in H9 c2 cardiac cells / 中国药理学通报

Aim To explore the role of nicotinamide adenine dinucleotide phosphate ( NADPH ) oxidase in high glucose-induced injury in H9c2 cardiac cells. Methods H9 c2 cardiac cells were exposed to differ-ent concentrations of high glucose(5. 5 mmol·L-1 ,11 mmol · L-1 ,22 mmol · L-1 ,33 mmol · L-1 ,44 mmol ·L-1 , 55 mmol · L-1 ) for 24 h and different time pints of high glucose(0 h,12 h,24 h,36 h,48 h,72 h) . Cell viability was measured by MTT colorimetry, the protein expression of Bcl-2 , Bax and NADPH oxi-dase submits such as p22 phox , p47 phox and p67 phox were determined by western blotting. Results H9c2 cardi-ac cells exposure to high glucose for 24 h showed on decrease in cell survival and the Bcl-2 expression while an increase in the Bax expression ( P<0. 05 ) . Moreo-ver, high glucose could markedly up-regulate the activ-ity of NADPH oxidase characterized by the enhanced expression of p22 phox , p47 phox and p67 phox ( P<0. 05 ) . Conclusion Activating NADPH oxidase may play an important role in high glucose-induced injury in H9 c2 cells.

Anti-apoptosis and anti-inflammatory effect of osthole in mice following stab wound injury / 中国药理学通报

Aim To investigate the effects of osthol on cell apoptosis and inflammatory cell infiltration after brain stab wound injury in mice. Methods The mice underwent the stab wound injury by a needle, then were randomly divided into sham operation group, model group, osthol 10, 20, 30 mg · kg-1 treatment group. The main examinations included mice brain wa-ter content; the apoptotic cytokines Bax, Bcl-2, Caspase-3 mRNA expression were assessed by PT-PCR; immunohistochemistry staining was used to de-tect neutrophils (MPO) and microglia (Iba-1) infiltra-tion and Caspase-3 positive cell expression around in-jured lesions. Results Treatment with osthole 20, 30 mg·kg-1 group significantly reduced the water content in injured brain, improved the ratio of Bax/Bcl-2, and reduced the expression of apoptosis cytokine Caspase-3 mRNA. Osthole 30 mg·kg-1 treatment group obvious-ly reduced the infiltration of neutrophils and microglial cells and significantly reduced the number of apoptotic cells around the injured cerebral cortex. Conclusion Osthole has therapeutic effect on stab wound injury in mice, and the possible mechanism may be by reducing the infiltration of inflammatory cells and reducing apop-totic cells.

Research progress on anti-tumor metastasis of cryptotanshinone / 中国药理学通报

Cryptotanshinone (CPT)is a major fat-soluble ingre-dient in Salvia,which is a traditional blood-activating and stasis-dissolving drug.CPT has been gradually concerned,because it has a remarkable therapeutic effect on cardiovascular diseases, cancer and neurodegenerative diseases.A large number of exper-imental and clinical studies have shown that CPT can primarily inhibit tumor cell′s proliferation,angiogenesis,invasion and ad-hesion and induce apoptosis.Thus to some extent,it hinders in-vasion of tumor cell and prevents the distant metastasis.This pa-per focuses on the anti-tumor metastasis of CPT.

Epidermal growth factor receptor-containing exosomes induce tumor-specific regulatory T cells / 中国药理学通报

Aim Toinvestigatewhetherepidermal growth factor receptor ( EGFR )-containing exosomes could induce tumor-specific regulatory T cells, and the effects of those T cells on tumor protein-specific CD8+Tcells.Methods TheexosomeswithEGFRwerepu-rified from NSCLC tumor, which modulated tolerogenic property of DCs. Then the induced TolDCs generated tumor-specific Tregs, with which the tumor protein-specific CD8 + T cells were suppressed. Results 80%exosomeswereEGFRpositivefromLCpatients while less than 2% exosomes were EGFR positive from control lung tissue. After exposed to the exosomes in the culture for 7 days, the IDO+ DCs proportion was much higher than that in control group ( 80. 8% ± 3. 2% vs 65. 6% ± 6. 4%, P <0. 05 ) . The induced Tregs was also higher ( 24. 1% ± 5. 2% vs 4. 2% ± 2. 3%,P<0. 01 ) , which suppressed the proliferation of CD8+ T cells(5. 4% ± 0. 2% vs 86. 7% ± 9. 3%, P<0.01).Conclusion Thepurifiedexosomesin-duce tolerogenic DCs. Coculture of the tolerogenic DCs and Th0 cells generates the tumor antigen-specific reg-ulatory T cells. The Tregs could suppress the tumor an-tigen specific CD8+ T cells.

The effects of total flavonoids of astragalus on the apoptosis of vascular endothelial cells induced by serum of uremia patient / 国际检验医学杂志

Objective To explore the effect of total flavonoids of astragalus (TFA) on the apoptosis of endothelial cells induced by serum of uremia patient .Methods The serum of 22 healthy volunteers and 25 uremia patients receiving regularly hemodialysis were enrolled in the study .HUVECs were used as research objects ,which were divided into control group(adding serum of healthy people when cell synchronized) and uremia group (adding serum of uremia patient when cell synchronized ) .Low dose ,moderate dose and high dose group were prepared by adding 0 .5 ,1 .0 ,2 .0 mg/mL TFA respectively 6 h before cell synchronization .After 24 hours′culture since the serum were added ,the morphological change of endothelial cells were observed by microscopy ,proliferation activities were tested by using MTT ,SOD activities were tested by using xanthine oxidase method ,NO levels were measured by u-sing nitrate reductase colorimetric method ,DNA damage was detected by using comet assay ,the morphological change of apoptosis was observed by using TUNEL method .Results Compared with the control group ,the proliferation activity ,SOD activity ,NO lev-els were lower in uremia group(P< 0 .01) ,DNA tailing rate ,apoptosis index(AI) significantly increased (P<0 .01) .Compared with cells of uremia group ,cell proliferation activity of all the TFA intervention groups increased (P<0 .05) ,NO levels also in-creased (P<0 .01) .Compared with uremia group ,moderate and high dose group′s SOD activity increased (P<0 .05) ,DNA damage tailing rate decreased (P<0 .05) .Conclusion Total flavonoids of astragalus reduces apoptosis of HUVECs induced by serum of uremia patient ,the possible mechanism is associated with the decrease of oxidative stress .

Study on the apoptosis of rat glioma cells induced by defective interfering particles of Sendai virus strain Tianjin / 中华微生物学和免疫学杂志

Objective To investigate the apoptosis of rat glioma C 6 cells induced by defective in-terfering( DI) particles of Sendai virus strain Tianjin .Methods Rat glioma C6 cells were treated with dif-ferent titers of DI particles of Sendai virus strain Tianjin in vitro with culture media as negative control and intact virus as positive control .At different time point , cells were collected and their apoptosis was detected by DNA gel electrophorsis , TUNEL assay and AnnexinⅤ/PI double-labeled flow cytometry .The C6 glioma-bearing rat model was established and then treated with three intratumoral injections of DI particles , intact virus or saline three times at interval of two days .The antitumor effects of ID particles were evaluated through daily measuring of the tumor size .Hematoxylin-eosin( HE) staining was used to observe the patho-logical changes in tumor tissues .TUNEL assay was performed to detect the apoptosis of tumor tissues .Re-sults Rat glioma C6 cells treated with DI particles or intact virus in vitro showed typical DNA ladder pattern in agarose gel electrophoresis in a time-and dose-dependent manner .With the intervention of DI particles , the apoptosis rate of C6 cells showed a time-and dose-dependent manner and was significantly higher than that of the control group (P0.05).Conclusion The DI particles of Sendai virus strain Tianjin could induce apoptosis of rat glioma C 6 cells in a time-and dose-dependent manner both in vitro and in vivo, suggesting that the DI particles might be applicable for the treatment of neurogliocytoma in the future.