Research progress of the regulatory role of autophagy in metabolic liver diseases / 中华肝脏病杂志

Autophagy is one of several hepatic metabolic processes in which starved cells are supplied with glucose, free fatty acids, and amino acids to produce energy and synthesize new macromolecules. Moreover, it regulates the quantity and quality of mitochondria and other organelles. As the liver is a vital metabolic organ, specific forms of autophagy are necessary for maintaining liver homeostasis. Protein, fat, and sugar are the three primary nutrients that can be altered by different metabolic liver diseases. Drugs that have an effect on autophagy can either promote or inhibit autophagy, and as a result, it can either increase or inhibit the three major nutritional metabolisms that are affected by liver disease. Thus, this opens up a novel therapeutic option for liver disease.

Regulative effect of Rhubarb dispensing granule on autophagy and gastrointestinal motility of Cajal interstitial cells in rats with chronic transit constipation / 国际中医中药杂志

Objective:To study the regulative effect of Rhubarb dispensing granule on autophagy and gastrointestinal motility of Cajal interstitial cells in rats with chronic transit constipation (STC).Methods:A total of 75 rats were randomly divided into control group, model group, low, medium and high dose groups with 15 rats in each group. Except for the control group, the STC rat models were established by intragastric administration of compound diphenoxylate suspension. Rats in low, medium and high dose groups were given Rhubarb dispensing granule of 1, 2 and 4 g/kg by gavage respectively, while rats in control group and model group were given normal saline with the same volume by gavage, once a day for 14 consecutive days. Fecal moisture content and intestinal propulsion rate were measured. The serum levels of substance P (SP), vasoactive intestinal peptide (VIP), NO and NOS were detected by ELISA. The pathological changes of colon tissue were observed by HE staining. The c-kit level in colon tissue was detected by immunohistochemistry. The mRNA and protein levels of c-kit and Beclin1 in colon tissue were detected by PCR and Western blot.Results:Compared with the model group, the fecal moisture content, the carbon pushing distance and the intestinal pushing rate of the low, medium and high dose groups were significantly increased ( P<0.05), the serum SP level was increased ( P<0.05), the serum VIP, NO and NOS levels of the low, medium and high dose groups were decreased ( P<0.05), and the average expression score of c-kit in colon tissue of the low, medium and high dose groups was significantly increased ( P<0.05). The levels of c-kit mRNA (2.33 ± 0.35, 3.04 ± 0.17, 3.83 ± 0.23 vs. 0.61 ± 0.07) and protein (0.42 ± 0.06, 0.60 ± 0.07, 0.79 ± 0.08 vs. 0.22 ± 0.04)in colon tissue of rats in low, medium and high dose groups were increased ( P<0.05), and Beclin1 mRNA (4.17 ± 0.37, 3.35 ± 0.44, 1.05 ± 0.28 vs. 6.04 ± 0.31) and protein (0.76 ± 0.11, 0.57 ± 0.08, 0.43 ± 0.05 vs. 0.91 ± 0.06) were decreased ( P<0.05). Conclusion:Rhubarb dispensing granule could significantly increase the fecal moisture, intestinal motility rate, serum SP level and colonic tissue c-kit level, decrease serum VIP, NO, NOS level and colonic tissue Beclin1 level in rats with chronic transit constipation, and then inhibit autophagy of Cajal interstitial cells and regulate gastrointestinal motility in rats with chronic transit constipation.

Cyclic Stretch Induces Adhesion of VSMCs with Platelet-Derived Microparticles and the Role in Autophagy / 医用生物力学

Objective To investigate the effect of cyclic stretch on adhesion of vascular smooth muscle cells (VSMCs) with platelet-derived microparticles (PMPs), and the role of PMPs in VSMC autophagy. Methods Cyclic stretch with the magnitude of 5% (simulating physiological mechanical stretch) or 15% (simulating pathological mechanical stretch) was subjected to VSMCs in vitro by using FX-5000T cyclic stretch loading system, and the adhesion of PMPs in VSMCs was detected by using flow cytometry. Immunofluorescence was used to detect the expression of autophagy microtubule associated protein light chain 3 (LC3) after 24 h stimulation with PMPs. Western blotting was used to detect the expression of autophagy related protein (Atg) in VSMCs after 24 h stimulation by PMPs. Results Compared with 5% cyclic stretch, 15% cyclic stretch significantly increased the adhesion ability of VSMCs with PMPs. Immunofluorescence and Western blotting result revealed that PMPs stimulation significantly increased the expression of autophagy marker protein LC3 in VSMCs. Furthermore, the protein expressions of Atg5, Atg7 and Atg12 were all significantly increased in VSMCs stimulated with PMPs. Conclusions High cyclic stretch may enhance the autophagy of VSMCs by promoting the adhesion of PMPs, which will subsequently increase the expressions of Atg5, Atg7, Atg12 and LC3.

Study on mechanism of Compound Kushen Injection inducing autophagy to promote apoptosis of bladder cancer cells / 中草药

Objective: To investigate the effect of Compound Kushen Injection on proliferation, apoptosis, and autophagy of bladder cancer T24 cells, and to explore the molecular mechanisms involved, and provide theoretical basis for Compound Kushen Injection in clinical treatment of bladder cancer. Methods: The effect of Compound Kushen Injection on proliferation of bladder cancer cells was detected by CCK-8 method. Flow cytometry was used to detect the effect of Compound Kushen Injection on bladder cancer cell apoptosis. MDC staining was used to detect the effect of Compound Kushen Injection on autophagy of bladder cancer cells. Western blotting and immunofluorescence experiments were used to detect the effect of Compound Kushen Injection on key protein involved in apoptosis and autophagy of bladder cancer cells. Results: Compound Kushen Injection effectively inhibited the proliferation of bladder cancer T24 cells. Compound Kushen Injection induced bladder cancer cells to undergo apoptosis and autophagy. The results of Western blotting and immunofluorescence experiments showed that as the concentration of Compound Kushen Injection increased, the expressions of Bcl-2 and P62 in bladder cancer T24 cells were gradually decreased, and the expressions of Bax, cleaved-Caspase 3, Beclin1, and LC3Ⅱ were increased (P < 0.05), but Bcl-2/Bax ratio was decreased (P < 0.05). Conclusion: Compound Kushen Injection can inhibit the proliferation of bladder cancer T24 cells, regulate autophagy of bladder cancer T24 cells, and induce bladder cancer T24 cell apoptosis. Key words:

Down-regulation of Beclin1 inhibits the resistance of ovarian cancer A2780/DDP cells for the cisplatin / 中国肿瘤生物治疗杂志

@#[Abstract] Objective: To investigate the effect of Beclin1 knockdown on cisplatin resistance in ovarian cancer A2780 cells and its related mechanisms. Methods: The mRNA and protein expressions of Beclin1 in A2780 cells and drug resistant A2780/DDP cells were determined by qPCR and Western blotting. After transfection with Beclin1 siRNA, the sensitivity of A2780/DDP cells to cisplatin was detected by MTT assay; Cell clone formation and apoptosis were detected by the Colony formation assay and Flow cytometry assay, respectively; cell autophagy was monitored by monodansylcadaverin (MDC) staining. Furthermore, the protein levels of cell autophagy related proteins, lysosomal associated membrane protein Lamp-2 and Cathepsin B were detected by Western blotting. Results: The mRNA and protein expression levels of Beclin1 in cisplatin-resistant A2780/DDP cells were significantly higher than those in A2780 cells (all P<0.05). The expression of Beclin1 was significantly increased in A2780 cells after treated with cisplatin (P<0.05). Beclin1 knockdown promoted cisplatin induced apoptosis of A2780/DDP cells (P<0.05), inhibited autophagy and cell colony formation (all P<0.05), and increased cell sensitivity to cisplatin (P<0.05). Meanwhile, Western blotting showed that Beclin1 knockdown increased the protein levels of cleaved-caspase 3 and Cathepsin B in A2780/DDP cells, while down-regulated the protein expressions of Atg3, Atg7, LC3Ⅱ/Ⅰand Lamp-2 (all P<0.05). Conclusion: Beclin1 knockdown can improve the sensitivity of A2780/DDP cells to cisplatin, and the mechanism may be related to the inhibition of protective autophagy of cells by regulating the expressions of autophagy related proteins, and the regulation of lysosomes, thus further promoting cisplatin-induced apoptosis of drug-resistant cells.

Effect of electroacupuncture preconditioning of "Neiguan"(PC6) on myocardial LKB1/AMPK/PFK2 pathway in myocardial ischemia rats / 针刺研究

OBJECTIVE: To observe the effect of electroacupuncture (EA) preconditioning on the expression of liver protein kinase 1 (LKB1), adenosine 5'-monophosphate (AMP)-activated protein kinase (AMPK) and 6-phosphofructo-2-kinase (PFK2) in cardiomyocytes of rats with acute myocardial ischemia (AMI), so as to explore its mechanisms underlying cardioprotective effect. METHODS: Thirty male Wistar rats were randomly divided into sham-operation, model and EA pretreatment groups (n＝10 rats per group). The AMI model was established by ligation of the left anterior descending branch of coronary artery. Before modeling, EA preconditioning (2 Hz/15 Hz, 1 mA) was applied to bilateral "Neiguan"(PC6) for 30 min, once daily for 14 days. Histopathological changes of myocardium was observed by microscope after H．E. staining. The level of lactate dehydrogenase (LDH) in serum was detected by ELISA. The expression of autophagy-associated proteins and mRNAs as LKB1, AMPKa1, AMPKa2 and PFK2 were detected by Western blot and real-time PCR, respectively. RESULTS: Compared with the sham-operation group, serum LDH content, and expression levels of myocardial AMPKa2 and PFK2 proteins and mRNAs were significantly up-regulated (P<0.01), and those of LKB1 and AMPKa1 proteins and mRNAs were increased in the model group (P<0.05). Following the intervention, serum LDH were apparently down-regulated (P<0.01), and expression levels of myocardial LKB1, AMPKa1 and PFK2 proteins and mRNAs were apparently up-regulated (P<0.01), but that of AMPKa2 protein and mRNA was remarkably down-regulated in the EA group (P<0.01). H．E. staining showed cell swelling, disordered arrangement of myocardial fibers with obvious rupture, interstitial bleeding and inflammatory infiltration, which was relatively milder in the EA preconditioning group. CONCLUSION: EA pretreatment can trigger LKB1/AMPK/PFK2 signaling pathway in AMI rats, which may contribute to its cardioprotective effect against ischemic myocardial injury by activating autophagy of cardiomyocytes.　.

Effect of Shenling Baizhu San on Murine Model of IBD Induced by DSS in Mice by Regulating Autophagy / 中国实验方剂学杂志

Objective: To investigate the mechanism of Shenling Baizhu San (SLBZS) in treating dextra sulfate sodium (DSS)-induced inflammatory bowel disease (IBD) mice and its relationship with autophagy. Method: SPF BALB/c mice were randomly divided into control group,model group,mesalazine group,low,medium and high-dose SLBZS groups,and autophagy inducer rapamycin group.The IBD mice were fed with 5% DSS in their drinking water for 7 days,and the control mice received only water.SLBZS groups were given SLBZS at doses of 3,6,12 g·kg-1·d-1, positive group was given mesalazine sustained release granules at the dose of 2 g·kg-1·d-1, rapamycin group was given rapamycin at the dose of 4 mg·kg-1·d-1, and control mice was given the same volume of normal saline by gavage.The mice weight,stool occult blood in stool,score of disease activity (DAI),pathological examination of intestinal mucosal lesions integral were observed after 7 days. interleukin(IL)-8 and IL-10 in serum were detected by enzyme\|linked immuno sorbent assay(ELISA), vascular tissue samples were prepared for the detection of tumor neorosis factor-(TNF-α) and IL-1β, and transmission electron microscope and Western blot were used to detect the formation of autophagosomes and the level of autophagy. Result: The body mass decrease, the colon length, disease activity scoring, and histological scoring of SLBZS group were better than those of DSS group. Compared with control group, the level of IL-10 decreased, while the level of IL-8 increased obviously (Pα expressions significantly up-regulated(PPPα expressions(PConclusion: Shenling Baizhu San can significantly inhibit the IBD by regulating autophagy and suppressing inflammation.

Protective effect of ceftriaxone on hippocampal neurons in subarachnoid hemorrhage rats and its mechanism / 吉林大学学报(医学版)

Objective: To investigate the therapeutic effect of ceftriaxone in the rats with subarachnoid hemorrhage (SAH), and to clarify its mechanism. Methods: A total of 48 adult male SD rats were randomly divided into sham operation group, SAH group, 3-methyl adenine (3-MA) group and ceftriaxone (CEF) group; there were 12 rats in each group. The SAH models were established by intravascular puncture; the rats were administered intraperitoneally; the dose of 3-MA was 15 mg · kg-1, and the dose of CEF was 500 mg · kg-1. The rats were sacrificed 24 h after model establishment, and the pathology of brain tissue of the rats was observed by HE staining. The apoptosis of neurons was detected by TUNEL staining. The number of autophagy-related proteins Beclin-1 and LC3-II and the apoptosis factor Caspase-3 protein in hippocampus tissue of the rats in various groups were detected by immunohistochemistry and Western blotting methods. Results: Compared with SAH group, the neurological score of the rats in CEF group was significantly increased (P<0. 01). The HE staining results showed the nerve cells in the hippocampus area of the rats in SAH group were swollen with loose cytoplasm and had obvious nuclear condensation, fragmentation and dissolution; compared with SAH group, the neural degeneration of the rats in CEF group was improved, and more normal nerve tissue morphology appeared; while the nerve cell damage in 3-MA group was more serious, the dead nerve cells were in full filed, and the cell layers were disordered. The quantitative analysis results showed that the number of necrotic nerve cells in CEF group was significantly lower than that in SAH, and the number of necrotic nerve cells in 3-MA group was significantly higher than that in SAH group (P< 0. 05). The immunohistochemical staining results showed that the number of Beclin-1 and LC3-II posivive cells in hippocampus tissue of the rats in CEF group was significantly higher than that in SAH group (P<0. 05); the number of Beclin-1 and LC3-II positive cells in 3-MA group was significantly lower than that in SAH group (P< 0. 05). The TUNEL staining resluts showed that the number of apoptotic nerve cells in CEF group was significantly lower than that in SAH group (P<0. 05), and the number of apoptotic nerve cells in 3-MA group was higher than that in SAH group (P<0. 05). The Western blotting results showed that the expression level of Caspase-3 protein in hippocampus tissue of the rats in CEF group was significantly lower than that in SAH group (P<0. 05), while the expression level of Caspase-3 protein in 3-MA group was significantly higher than that in SAH group (P< 0. 05). Conclusion: CEF has the protective effect on the nerve injury of the rats with SAH, and its mechanism may be related to up-regulation of autophagy and reduction of apoptosis of the neurons.

Advances in the regulation of PI3K signaling pathway on acinar cells in acute pancreatitis / 解放军医学杂志

Acute pancreatitis (AP) is a systemic inflammatory disease with complex pathogenesis. Acinar cell injury is the earliest pathological change of AP. Elucidating its pathological mechanism is of great significance for the development of new therapeutic strategies for AP. Studies have shown that activation of PI3K/Akt signaling pathways plays an important role in the progression of acute pancreatitis. It can not only contribute to acinar cell necrosis by inhibiting apoptosis, but also regulate the autophagy, oxidative stress and inflammation of acinar cells. Here, we reviews the advances about the PI3K/Akt signaling pathway and its significance in modulation of acinar cells in acute pancreatitis.

Effects of Aspirin on the Growth and Autophagy of Human Gastric Cancer Cells SGC- 7901 and BGC- 823 / 中国药房

OBJECTIVE： To study the effects of aspirin on the growth and autoghagy of human gastric cancer cells SGC-7901 and BGC-823. METHODS： SGC-7901 and BGC-823 cells were selected as research objects， with phosphate buffer （PBS） as negative control treated for 48 h， MTT assay was used to detect the effects of 1， 2， 4， 6， 8， 10 mmol/L aspirin， 5 mmol/L aspirin alone or combined with 2.5 μmol/L chloroquine， 2.5 μmol/L 3-methyladenine （3-MA） on survival rate of gastric cancer cells. Flow cytometry was used to detect the effects of 2 and 5 mmol/L aspirin， 5 mmol/L aspirin alone or combined with 2.5 μmol/L chloroquine and 2.5 μmol/L 3-MA on the apoptosis rate and cell cycle distribution of gastric cancer cells. Hoechst33258 staining was used to observe the effects of 5 mmol/L aspirin on morphology of gastric cancer cell nucleus； Transwell chamber test was adopted to detect the effects of 5 mmol/L aspirin on the migration of gastric cancer cell. Laser confocal scanning microscopy was used to observe the effects of 5 mmol/L aspirin on autophagy formation in gastric cancer cells. Western blot method was used to detect the effects of 2 and 5 mmol/L aspirin on the protein expression of autophagy markers LC3-Ⅱin gastric cancer cells. RESULTS： Compared with negative control group， aspirin could inhibit the survival rates of SGC-7901 and BGC-823 cells in dose-dependent manner， but had no significant effects on apoptosis rate of SGC-7901 and BGC-823 cells； SGC-7901 and BGC-823 cells were blocked in G1 phase. Compared with aspirin alone group， the survival rates of SGC-7901 and BGC-823 were increased significantly after treated with aspirin+chloroquine and aspirin+3-MA， while the distribution rate of SGC-7901 and BGC-823 cells at G1 phase were decreased significantly， with statistical significance （P＜0.05 or P＜0.01）. Compared with negative control group， there were no obvious DNA fragmentation fragments， apoptotic bodies and fragments of dense bright blue， while the number of migration cells were decreased significantly in SGC-7901 and BGC-823 cells after treated with aspirin （P＜0.001）； the number of autophagosome was increased significantly and the protein expression of LC3-Ⅱ was enhanced significantly （P＜0.05）. CONCLUSIONS： Aspirin can significantly inhibit the growth of SGC-7901 and BGC-823 cells， and arrest cell cycle in G1 phase， the mechanism of which may be associated with the activation of autophagy.

Kaempferol suppresses human gastric cancer SNU-216 cell proliferation, promotes cell autophagy, but has no influence on cell apoptosis

Gastric cancer remains a serious threat to human health worldwide. Kaempferol is a plant-derived flavonoid compound with a wide range of pharmacological activities. This study aimed to investigate the effects of kaempferol on gastric cancer SNU-216 cell proliferation, apoptosis, and autophagy, as well as underlying potential mechanisms. Viability, proliferation, and apoptosis of SNU-216 cells after kaempferol treatment were evaluated using cell counting kit-8 assay, 5-btomo-2′-deoxyuridine incorporation assay, and annexin V-FITC/PI staining, respectively. Quantitative reverse transcription PCR was performed to measure the mRNA expressions of cyclin D1 and microRNA-181a (miR-181a) in SNU-216 cells. Cell transfection was used to down-regulate the expression of miR-181a. The protein expression levels of cyclin D1, bcl-2, bax, caspase 3, caspase 9, autophagy-related gene 7, microtubule-associated protein 1 light chain 3-I (LC3-I), LC3-II, Beclin 1, p62, mitogen-activated protein kinase (MAPK), extracellular regulated protein kinases (ERK), and phosphatidylinositol 3 kinase (PI3K) in SNU-216 cells were detected using western blotting. Results showed that kaempferol significantly suppressed SNU-216 cell viability and proliferation but had no influence on cell apoptosis. Further results suggested that kaempferol significantly induced SNU-216 cell autophagy. The expression of miR-181a in SNU-216 cells after kaempferol treatment was enhanced. Kaempferol significantly inactivated MAPK/ERK and PI3K pathways in SNU-216 cells. Suppression of miR-181a significantly reversed the kaempferol-induced MAPK/ERK and PI3K pathways inactivation in SNU-216 cells. This research demonstrated that kaempferol suppressed proliferation and promoted autophagy of human gastric cancer SNU-216 cells by up-regulating miR-181a and inactivating MAPK/ERK and PI3K pathways.

Moxibustion improves cardiac function by up-regulating autophagy-related proteins of cardiomyocytes in rats with chronic heart failure / 针刺研究

OBJECTIVE: To observe the effect of moxibustion on cardiac function and the expression of autophagy-related proteins microtubule-associated protein 1 light chain 3 (LC3) and selective autophagy receptor signaling adaptor sequestosome 1 (SQSTM1/p62) in rats with chronic heart failure (CHF), so as to explore its underlying mechanisms in preventing and treating CHF. METHODS: Sixty male SD rats were randomly divided into normal, model, moxibustion, autophagy inhibitor 3-methyladenine (3-MA) and autophagy agonist rapamycin (RAPA) groups (n＝12 rats/group). The CHF model was established by intrape-ritoneal injection of adriamycin (ADR, 2 mg/kg, once every week for 12 weeks). Mild moxibustion was applied to bilateral "Feishu" (BL13) and "Xinshu" (BL15) for 20 min every time. Rats of the 3-MA group were treated by intraperitoneal injection of 3-MA suspension (15 mg/kg), and those of the RAPA group treated by gavage of RAPA suspension (2 mg/kg). All the treatments were given once a day for 3 weeks. The heart rate (HR), cardiac output (CO), left ventricular systolic pressure (LVSP), left ventricular end-diastolic pressure (LVEDP) and maximum rising and lowering rates of left ventricular pressure (±dp/dtmax) were measured for assessing the cardiac performance. Histopathological changes of the left ventricular myocardium were observed by HE staining. The expression levels of LC3-Ⅰ， LC3-Ⅱ and p62 proteins of the left ventricle myocardium tissue were detected by Western blot. RESULTS: After modeling, the pathological changes of myocardium (as myocardial cell swelling with vacuoles, myocardial fibre breakage, etc.) were obvious, and the HR, LVEDP, LC3-Ⅱ and LC3-Ⅱ/Ⅰ protein expression levels were significantly increased in the model group compared with the normal group (P<0.01), while the CO, LVSP, ±dp/dtmax, and the expression of p62 protein were significantly down-regulated (P<0.01). Following the interventions, the myocardial injury was reduced, the HR, LVEDP, LC3-Ⅱ and LC3-Ⅱ/Ⅰ levels in both moxibustion and 3-MA groups were significantly decreased (P<0.05, P<0.01), while the CO, LVSP, ±dp/dtmax and p62 expression level were significantly increased relevant to the model group (P<0.05, P<0.01). In addition, the ratio of LC3-Ⅱ/Ⅰ was significantly increased, and the expression level of p62 significantly down-regulated in the RAPA group compared with the model group (P<0.01). CONCLUSION: Moxibustion can improve cardiac function in CHF rats, which may be related to its effects in down-regulating the ratio of LC3-Ⅱ/Ⅰ and up-regulating the expression of p62 protein to inhibit cardiomyocyte autophagy.

Mechanisms and effects of PM2.5 on diseases of respiratory system: Latest findings / 西安交通大学学报(医学版)

The chemical composition of PM2.5 is extremely complex and it can carry some viruses and bacteria, gather many toxic and hazardous substances. The respiratory system is the main target organ of PM2.5 exposure and impact. There are many different kinds of respiratory system diseases, which have complex etiologies and unpredictable outcomes. Previous reports have shown that PM2.5 plays an important role in the occurrence and development of respiratory obstructive, infectious, allergic and neoplastic diseases; of which mechanisms may be related to oxidative stress, inflammatory reaction, genetic toxicity, cell apoptosis, cell autophagy and cell cycle disorders.

Significance of cell autophagy in resorption of lumbar disc herniation / 中国骨伤

As a self-protective mechanism of cells, autophagy of cells can maintain cell stability by degrading self-aging substances, and it can be highly induced. The ability of autophagy to degraded cells will decrease with age. The resorption phenomenon after lumbar disc herniation is one of the effective mechanisms in conservative treatment of lumbar disc herniation. The degenerative lesion of intervertebral disc is one of the main reasons of lumbar disc herniation. Cell autophagy is extensive participation in the degeneration of lumbar intervertebral disc, delaying the occurrence of degenerative disease. Futhermore, cell autophagy can potentially induce the occurrence of reabsorption. The study of cell autophagy has great significance to the degenerative disease of intervertebral disk and the reabsorption of lumbar disc herniation. And it is also of great significance for the clinical treatment of patients with lumbar disc herniation. For this reason, we should pay more attention to the study of cell autophagy in resorption.

Cell autophagy attenuates PM2.5-induced apoptosis of human lung adenocarcinoma cell line H441 / 基础医学与临床

Objective To investigate the function of autophagy in the process of PM2.5-induced apoptosis. Methods PM2.5 was obtained from Zhanjiang in 2016. Human lung adenocarcinoma cells H441 were treated with PM2.5 at different concentrations for different times. Cell proliferation was analyzed by MTT assay; Cell apoptosis was assessed by PI and Annexin V double staining and TUNEL assay. The expression of autophagy marker LC3Ⅱ, AKT and P-AKT protein was examined by Western blot ( WB). H441 cells were treated with PM2.5 following treatment with rapamycin or 3-MA. Cell viability was evaluated by trypan blue staining. Results Compared with the control group, cell proliferation was significantly inhibited by PM2.5 at concentration of 100 μg/mL or more for 24 and 48 h. With the increase of PM2.5 concentration, the cells apoptotic rate significantly increased, the protein ex-pression of LC3Ⅱwas increased as well as the P-AKT was decreased; and the protein expression of LC3Ⅱwas in-creased significantly after AKT inhibitor treatment. Moreover, rapamycin decreased PM2.5-induced cell apoptosis, and 3-MA can promote PM2.5-induced cell apoptosis. Conclusions In H441 cells, PM2.5 activates autophagy by inhibiting activation of AKT pathway, and cell autophagy can mitigate PM2.5-induced apoptosis.

Relationship between Autophagy and Repair of Spinal Cord Injury Based on Pathogenesis of Stasis Obstructing Meridian (review) / 中国康复理论与实践

Spinal cord injury is a kind of central nervous system injury disease caused by trauma. It was found that the up-regulation of autophagy after spinal cord injury could promote the repair of spinal cord injury. Under the condition of stress, autophagy can reduce the number of misfolded proteins and damaged organelles in cells to maintain the stability of intracellular environment. Chinese medicine with Huoxue Quyu Tongluo (against stasis obstructing meridian) efficacy has attracted much attention in the treatment of spinal cord injury, which may associate with the role of regulation of autophagy.

Regulating effects of miR-144 on Beclin-1 gene expression in ovarian cancer cells SKOV-3 / 基础医学与临床

Objective To detect the influence of rapamycin on the expression of 4 kinds of miRNAs and the effect cell autophagy.To study the relationship of miR-144 and Beclin-1 gene.Methods SKOV-3 cells were treated with 50 ng/mL rapamycin 2 hours and 10 nmol/L 3-methyl adenine 12 hours,the expression of miR-17,miR-20a,miR-144 and miR-155 was detected by RT-qPCR in SKOV-3 cell of different groups,the protein expression of Beclin-1 was detected by Western blot.The targeting effect of miR-144 on Beclin-1 gene was verified by the dual-luciferase reporter assay,Western blot and RT-qPCR.Results The expression of miR-17,miR-144 and miR-155 were in creased compared with NC groups in rapamycin group (P＜0.05);miR-17,miR-20a and miR-144 were down regulated compared with NC group in 3-MA group(P＜0.05);the protein of Beclin-1 was down expression compared with NC group in rapamycin group.miR-144 could suppress Beclin-1 expression by targeting the specific 3'untranslated region sequence of Beclin-1 gene.Conclusions miR-144 can inhibit the autophagy-related gene Beclin-1 expression and regulate the autophagy process in SKOV-3 cells.

Virological characteristics of Zika virus and abnormal immune responses induced by Zika virus / 中华微生物学和免疫学杂志

Zika fever is a self-limiting and acute infectious disease caused by Zika virus infection. It is mainly transmitted through the bite of mosquitoes of the Aedes type. It can also be spread through verti-cal transmission. There is evidence that it can also be sexually transmitted from a man to his sex partners due to the presence of the virus in semen. The re-emergence of the virus in 2015 as a major endemic in the South American countries ( which may spread further) warrants better understanding of Zika virus. The outbreaks, transmission routes, virological characteristics and the diagnosis and treatment of Zika virus infection will be summarized in this review. Moreover, the potential correlations between newborn microcephaly and Zika vi-rus infection as well as the possible molecular mechanisms for causing microcephaly such as cell autophagy will also be discussed.

Pterostilbene induces retinoblastoma WERI-Rb-1 cell apoptosis via autophagy induction / 中国癌症杂志

Background and purpose:Pterostilbene is a natural antioxidant, whose role in retinoblastoma remains unclear. The aim of this study is to probe the effects of pterostilbene on the proliferation, apoptosis and autophagy in retinoblastoma WERI-Rb-1 cell lines.Methods:Cell counting kit-8 (CCK-8) assays were used to analyze the effects of pterostilbene on the proliferation of WERI-Rb-1 cells. Apoptosis rate was determined by Annexin V/PI. Autophagic vacuoles were observed by acridine orange staining. LC3 and P62 protein expressions were determined using Western blot.Results:Pterostilbene significantly inhibited the proliferation of WERI-Rb-1 cells (P<0.01). The cell viability were (93.02±0.47)%, (55.10±2.04)% and (30.33±1.45)% after WERI-Rb-1 cells were treated with 25, 50 and 100 μmol/L pterostilbene for 24 h, and the cell viability were (88.38±3.70)%, (53.37±1.17)%, (29.60±1.05)% after WERI-Rb-1 cells were treated with 50 μmol/L pterostilbene for 12, 24 and 48 h. Pterostilbene induced cell apoptosis (P<0.01), the apoptosis rates of control group, 24 h treated group and 48 h treated group were (4.08±0.79)%, (13.44±2.12)% and (23.49±2.01)%. Pterostilbene induced autophagy of WERI-Rb-1 cells, increased LC3 expression, downregulated P62 expression and increased the number of autophagic vacuoles in WERI-Rb-1 cells (P<0.01). 3-MA and Beclin1 were able to rescue pterostilbene-induced cell death (P<0.01). After 3-MA was used to blunt autophagosome formation, the apoptosis rate markedly decreased in 3-MA+pterostilbene-treated cells compared with cells treated with pterostilbene alone [(12.97±2.09)%vs (8.35±1.11)%], and after siRNA was used to knockdown Beclin1, the apoptosis rate had the same change [(13.80±2.19)%vs (9.62±0.52)%].Conclusion:Pterostilbene can inhibit the proliferation of WERI-Rb-1 cells and induce cell apoptosis via autophagy activation.

Blocking autophagy magnifies MK-2206-induced DNA damage in SGC-7901 cells / 中国病理生理杂志

AIM:To investigate the effect of MK-2206, an inhibitor of protein kinase B (Akt), on the DNA damage of SGC-7901 cells.METHODS: SGC-7901 cells were treated with different concentrations of MK-2206, and phosphorylated histone H2AX (γ-H2AX) foci formation was detected by immunofluorescence staining .Western blot analy-sis was used to exam the levels of DNA damage-related protein.The expression of LC3-Ⅱ was determined to evaluate the change of autophagy .RESULTS:MK-2206 treatment increased the formation of γ-H2AX foci and histone H2AX phospho-rylation in the SGC-7901 cells.The levels of DNA damage response protein were also increased .In addition, MK-2206-treated SGC-7901 cells increased the expression of LC 3-II, a hallmark of autophagy .Inhibition of autophagy significantly enhanced MK-2206-mediated histone H2AX phosphorylation.CONCLUSION:MK-2206 induces DNA damage and auto-phagy in SGC-7901 cells.Blocking autophagy potentiates the response of MK-2206-induced DNA damage .