Protective Effects of Danmu Extract Syrup on Acute Lung Injury Induced by Lipopolysaccharide in Mice through Endothelial Barrier Repair / 中国结合医学杂志

OBJECTIVE@#To investigate the effects of Danmu Extract Syrup (DMS) on lipopolysaccharide (LPS)-induced acute lung injury (ALI) in mice and explore the mechanism.@*METHODS@#Seventy-two male Balb/C mice were randomly divided into 6 groups according to a random number table (n=12), including control (normal saline), LPS (5 mg/kg), LPS+DMS 2.5 mL/kg, LPS+DMS 5 mL/kg, LPS+DMS 10 mL/kg, and LPS+Dexamethasone (DXM, 5 mg/kg) groups. After pretreatment with DMS and DXM, the ALI mice model was induced by LPS, and the bronchoalveolar lavage fluid (BALF) were collected to determine protein concentration, cell counts and inflammatory cytokines. The lung tissues of mice were stained with hematoxylin-eosin, and the wet/dry weight ratio (W/D) of lung tissue was calculated. The levels of tumor necrosis factor-α (TNF-α), interleukin (IL)-6 and IL-1 β in BALF of mice were detected by enzyme linked immunosorbent assay. The expression levels of Claudin-5, vascular endothelial (VE)-cadherin, vascular endothelial growth factor (VEGF), phospho-protein kinase B (p-Akt) and Akt were detected by Western blot analysis.@*RESULTS@#DMS pre-treatment significantly ameliorated lung histopathological changes. Compared with the LPS group, the W/D ratio and protein contents in BALF were obviously reduced after DMS pretreatment (P<0.05 or P<0.01). The number of cells in BALF and myeloperoxidase (MPO) activity decreased significantly after DMS pretreatment (P<0.05 or P<0.01). DMS pre-treatment decreased the levels of TNF-α, IL-6 and IL-1 β (P<0.01). Meanwhile, DMS activated the phosphoinositide 3-kinase/protein kinase B (PI3K/Akt) pathway and reversed the expressions of Claudin-5, VE-cadherin and VEGF (P<0.01).@*CONCLUSIONS@#DMS attenuated LPS-induced ALI in mice through repairing endothelial barrier. It might be a potential therapeutic drug for LPS-induced lung injury.

Effect of South African Herb Hoodia gordonii on Liver Glucolipid Metabolism and PI3K/Akt/FoxO1 Signalling Pathway in db/db Mice / 中国实验方剂学杂志

ObjectiveTo observe the effects of the South African herb Hoodia gordonii （HG） on glucolipid metabolism in diabetic db/db mice and explore the possible mechanisms of HG on the liver of db/db mice based on the phosphoinositide-3 kinase （PI3K）/protein kinase B （Akt）/factor forkhead protein O1 （FoxO1） signaling pathway. MethodA total of 30 db/db mice were randomly divided into five groups according to fasting blood glucose： model group， metformin group （0.195 g·kg-1）， and low dose （0.39 g·kg-1）， medium dose （0.78 g·kg-1）， and high dose （1.56 g·kg-1） HG groups， with six m/m mice in each group， and another six m/m mice were set as normal group. The mice in the normal and model groups were given saline of 9 mL·kg-1 by gavage. Body weight， water intake， and fasting blood glucose of the mice in each group were measured weekly. After six weeks of continuous administration， serum insulin （FINS）， low-density lipoprotein cholesterol （LDL）， total cholesterol （TC）， triglyceride （TG）， alanine aminotransferase （ALT）， aspartate aminotransferase （AST）， urea， and creatinine （CREA） were measured， and liver sections were embedded and stained with hematoxylin-eosin （HE）， periodic acid-Schiff （PAS）， and oil red O. Protein expression of PI3K p85， p-Akt， and p-FoxO1 in liver was detected by immunohistochemistry. The mRNA expression of PI3K， Akt， and FoxO1 in liver tissue was detected by real-time polymerase chain reaction （Real-time PCR）. ResultAfter six weeks of administration intervention， it was found that fasting blood glucose was significantly downregulated in mice in the three HG groups （P<0.05）. The level of islet resistance index was significantly reduced in both the low and medium dose HG groups （P<0.05）. The expression levels of TC， TG， and LDL were reduced in all HG groups （P<0.05， P<0.01）. Pathologically， HG could alleviate hepatocyte steatosis， reduce the volume and content of lipid droplets in liver， and increase the distribution of glycogen granules in liver to some extent in mice. Immunohistochemical assays revealed that PI3K p85 protein expression was significantly increased in the low， medium， and high dose HG groups compared with the model group （P<0.01）. p-Akt protein expression was significantly increased in the medium and high dose HG groups （P<0.05， P<0.01）. p-FoxO1 protein expression was significantly increased in the low， medium， and high dose HG groups （P<0.05， P<0.01）. Compared with the model group， PI3K mRNA was increased in low dose， medium dose， and high dose HG groups （P<0.05）， and Akt mRNA was increased in high dose HG group （P<0.05）. FoxO1 mRNA was decreased in low dose， medium dose， and high dose HG groups （P<0.05）. ConclusionHG can ameliorate the disorder of glucolipid metabolism in db/db mice， which may be related to its activation of the hepatic PI3K/Akt/FoxO1 signaling pathway.

Effect mechanism research of procyanidin on gingivitis rats by regulating the PI3K/Akt/VEGF signal pathway / 中国药房

OBJECTIVE To investigate the potential mechanism of procyanidin on rats with gingivitis by regulating phosphoinositide 3-kinase （PI3K）/protein kinase B （Akt）/vascular endothelial growth factor （VEGF） signaling pathway. METHODS The rat model of gingivitis was constructed by sewing the neck of the first maxillary molar with silk thread+applying maltose on the gum+feeding with 20% sucrose solution and soft food. Forth-eight model rats were randomly divided into model group， procyanidin group （160 mg/kg）， 740Y-P group （PI3K/Akt signaling pathway activator， 0.02 mg/kg）， and procyanidin+ 740Y-P group （procyanidin 160 mg/kg+740Y-P 0.02 mg/kg）， with 12 rats in each group； another 12 rats were selected as control group； each medication group was treated with corresponding drugs intragastrically or/and intraperitoneally， once a day， for 7 consecutive days. Twenty-four hours after the last administration， the gingival index of rats was measured； the levels of interleukin- 18 （IL-18）， inducible nitric oxide synthase （iNOS） and alkaline phosphatase （ALP） in gingival crevicular fluid， as well as the levels of superoxide dismutase （SOD）， catalase （CAT） and reactive oxygen species （ROS） in gingival tissues of rats were detected； the pathological changes in gingival tissues were observed； the expression levels of PI3K/Akt/VEGF signaling pathway- related proteins in gingival tissues of rats were detected. RESULTS Compared with control group， the gingival tissues of rats in the model group had severe pathological damage，which was manifested as local tissue expansion and congestion， new capillaries， degeneration and loss of collagen fibers and disorder of arrangement， and a large number of inflammatory cell infiltration in the gingival sulcus wall. The gingival index， the levels of IL-18， iNOS， ALP in gingival crevicular fluid， the level of ROS in gingival tissues， the phosphorylations of PI3K and Akt， as well as the protein expression of VEGF in gingival tissues were significantly increased； the levels of SOD and CAT in gingival tissues of rats in model group were significantly decreased （P＜0.05）. Compared with model group， the pathological damage to the gingival tissues of rats in procyanidin group was reduced， and all quantitative indicators were significantly improved （P＜0.05）； 740Y-P could reverse the improvement effect of procyanidin on various indicators （P＜0.05）. CONCLUSIONS Procyanidin may alleviate gingival tissue damage， and improve gingival inflammation and oxidative stress in rats with gingivitis by inhibiting PI3K/Akt/VEGF signaling pathway.

Effect of hydrogen-rich water on brain injury in rats with acute radiation based on PI3K/AKT/Caspase-9 signaling pathway / 环境与职业医学

Background At present, radiation therapy is widely used in clinical treatment of tumors. However, while radiation therapy damages tumor cells, it also injures surrounding normal tissues. Studies have shown that hydrogen is a potential radiation-protective agent. Objective To investigate the neuroprotective mechanisms of hydrogen-rich water activating phosphoinositide 3-kinase (PI3K)/protein kinase B (AKT)/cysteinyl aspartate specificproteinase-9 (Caspase-9) signaling pathway in acute radiation-induced brain injury. Methods Forty male SD rats were randomly divided into four groups: control group, irradiation only group (IR), high-dose hydrogen-rich water intervention group (IR+HHRW), and low-dose hydrogen-rich water intervention group (IR+LHRW), 10 rats in each group. Except for the control group, animals in each group received a single 20 Gy whole brain irradiation. Animals in all groups were gavaged once a day from 3 d before irradiation to 7 d after irradiation, pure water (20 mL·kg−1) was given to the control and the IR groups, and hydrogen-rich water (20 mL·kg−1, 10 mL·kg−1) was given to the IR+HHRW and the IR+LHRW groups. After 7 d of intervention, 5 rats in each group were selected for the Morris water maze experiment for behavioral evaluation. Autopsies were conducted after anesthesia for the remaining animals and blood samples were collected for hematological analysis. Rat brains were harvested for TUNEL staining to observe neuronal apoptosis. HE staining was performed to observe histopathological changes, enzyme-linked immunosorbent assay was adopted to detect oxidative stress-related indicators, and real-time PCR and Western blotting were used to measure the expressions of PI3K/AKT/Caspase-9 pathway-related genes and proteins. Results The body weight of rats receiving irradiation decreased after 7 d of irradiation compared with the control group (P<0.05), and the symptoms such as arched back and malaise occurred to varying degrees, and the symptoms of rats in the IR+HHRW group were significantly milder than those in the IR group. The behavioral test results showed that the escape latency of rats in the IR+HHRW group or the IR+LHRW group was shorter than that in the IR group from day 2 to day 5 (P<0.05), and it took less time for rats in the IR+HHRW group to reach the original position after removing the platform on day 6 (P<0.05). The hematological test results showed that red blood cell (RBC) count, hemoglobin (HGB) level, and white blood cell (WBC) count were significantly decreased in the IR group (P<0.05), and the changes in the IR+HHRW group were improved (P<0.05). The HE staining results showed that the number of abnormal nerve cells, broken and dissolved nuclei, and the degree of damage in the IR+HHRW group were significantly reduced than those in the IR group. The results of oxidative stress evaluation showed that the ability of the IR group to inhibit free radicals decreased, the level of malondialdehyde (MDA) increased (P<0.01); the MDA level decreased after LHRW intervention (P<0.05); the SOD activity was elevated after HHRW intervention (P<0.05). The TUNEL staining results showed that the apoptosis signals in the IR+HHRW group were sparser than those in the IR group (P<0.05). The real-time PCR results showed that compared with the IR group, the mRNA expression levels of PI3K and AKT in the IR+HHRW group and the IR+LHRW group increased (P<0.05), while the mRNA expression levels of Cytc and Caspase-9 decreased (P<0.05). The Western blotting results showed that compared with the IR group, the phospho-AKT (pAKT) protein expression level in the IR+HHRW group increased significantly (P<0.05), while the expression of Caspase-9 and Cytc proteins decreased significantly (P<0.05). Conclusion Hydrogen-rich water can significantly reduce inflammation and oxidative stress caused by acute irradiation-induced brain injury, and decrease neuronal apoptosis. The mechanism may be related to the PI3K/AKT/Caspase-9 signaling pathway.

Mechanism of Buyang Huanwutang Combined with Bone Marrow Mesenchymal Stem Cell Transplantation in Treatment of Spinal Cord Injury Based on PI3K/Akt Signaling Pathway / 中国实验方剂学杂志

ObjectiveTo explore the mechanism of Buyang Huanwutang combined with bone marrow mesenchymal stem cell （BMSC） transplantation in the treatment of spinal cord injury （SCI）. MethodDifferent concentrations （12.5， 25， 50 g·kg-1） of Buyang Huanwutang were administrated to rats by gavage. The spinal cord function of rats was measured by modified Tarlov score， and the most suitable concentration of Buyang Huanwutang was screened out. SD rats were then divided into 6 groups， namely， the sham operation group （gavage of equal amount of normal saline）， the model group （gavage of equal amount of normal saline）， the Buyang Huanwutang group （gavage of 25 g·kg-1 Buyang Huanwutang）， the BMSC transplantation group （tail vein injection of BMSCs 1 mL）， the Buyang Huanwutang+BMSC group （gavage of 25 g·kg-1 Buyang Huanwutang and tail vein injection of BMSCs 1 mL）， the Buyang Huanwutang+BMSC+LY294002 group （gavage of 25 g·kg-1 Buyang Huanwutang and tail vein injection of BMSCs 1 mL and 40 mg·kg-1 LY294002）， with 10 rats in each group. The spinal cord function was measured by the modified Tarlov score， inclined plate test， and latency of cortical somatosensory evoked potential. Immunohistochemistry was used to detect the number of 5-bromo-2-deoxyuracil nucleoside （Brdu）-labeled positive cells in the spinal cord tissue. The protein expression levels of phosphorylated protein kinase B （p-Akt）， glycoprotein 130 （gp130）， and interleukin-6 （IL-6） in spinal cord were detected by Western blot. ResultAs compared with the sham operation group， the Tarlov score and the critical angle of tilt plane in the model group were significantly decreased （P<0.05）， and the latency of cortical somatosensory evoked potential wave and the protein expression levels of p-Akt， gp130， and IL-6 were significantly increased （P<0.05）. As compared with the model group， the Tarlov score and the critical angle of tilt plane in the sham operation group and each treatment group were significantly increased （P<0.05）， and the latency of cortical somatosensory evoked potential wave and the protein expression levels of p-Akt， gp130， and IL-6 were significantly decreased （P<0.05）. As compared with the BMSC group， the Tarlov score and the critical angle of inclined plane in the Buyang Huanwutang+BMSC group increased （P<0.05）， the latency of cortical somatosensory evoked potential wave and the protein expression levels of p-Akt， gp130， and IL-6 decreased （P<0.05）， and the number of Brdu-labeled positive cells increased 5 weeks after transplantation （P<0.05）. As compared with the Buyang Huanwutang+BMSC group， the Tarlov score and the critical angle of the inclined plane in the Buyang Huanwutang+BMSC+LY294002 group increased （P<0.05）， and the latency of cortical somatosensory evoked potential wave and the protein expression levels of p-Akt， gp130， and IL-6 decreased significantly （P<0.05）. Five weeks after transplantation， the number of Brdu-labeled positive cells increased significantly in the Buyang Huanwutang+BMSC+LY294002 group （P<0.05）. ConclusionBuyang Huanwutang can promote BMSCs migration and restore spinal cord function by inhibiting phosphatidylinositol 3-kinase （PI3K）/protein kinase B （Akt） signal.

Inhibitory effects of formononetin on lipopolysaccharide-induced apoptosis and inflammatory response in alveolar epithelial cells / 中国药房

OBJECTIVE To investigate the inhibitory effects of formononetin on lipopolysaccharide （LPS）-induced apoptosis and inflammatory response in alveolar epithelial cells through phosphoinositide 3-kinase （PI3K）/protein kinase B （Akt） signaling pathway. METHODS Human lung cancer alveolar basal epithelial cells A549 were cultured in vitro and divided into control group （no intervention）， model group （1 μg/mL LPS）， different concentrations of formononetin groups （1 μg/mL LPS+6.25， 12.5， 25， 50 μmol/L formononetin）. The levels of inflammatory factors （interleukin-8， tumor necrosis factor-α） and cell viability were detected in each group. Another A549 cells were divided into control group， model group （1 μg/mL LPS）， LPS+25 group （1 μg/mL LPS+25 μmol/L formononetin）， inhibitor group （1 μg/mL LPS+20 μmol/L LY294002）， formononetin+inhibitor group （1 μg/mL LPS+25 μmol/L formononetin+20 μmol/L LY294002） and formononetin+activator group （1 μg/mL LPS+25 μmol/L formononetin+ 10 μmol/L SC79）. The secretion levels and mRNA expressions of inflammatory factors， cell apoptosis， and expressions of the key proteins of PI3K/Akt signaling pathway were detected in each group. RESULTS Compared with model group， the levels of inflammatory factors were decreased significantly after the intervention of 25 μmol/L of formononetin， and the cell viability was increased significantly （P＜0.05）. Compared with the control group， the secretion levels and mRNA expressions of inflammatory factors， apoptotic rate， and relative expressions of phosphorylated Akt and phosphorylated PI3K of the model group were increased significantly （P＜0.05）. Compared with the model group， the above indexes of the LPS+25 group and the inhibitor group were decreased significantly （P＜0.05）. Compared with the LPS+25 group， the above indicators of formononetin+inhibitor group were further decreased， while those of formononetin+activator group were increased significantly （P＜0.05）. CONCLUSIONS Formononetin can inhibit LPS-induced epithelial cell apoptosis and improve inflammatory response， and the mechanism may be related to the inhibition of the PI3K/Akt signaling pathway.

Effect of Shengjiang Tonglong Prescription Hollow Suppository on Rats with Prostatic Hyperplasia Based on PI3K/Akt Signaling Pathway / 中国实验方剂学杂志

ObjectiveTo investigate the effect of Shengjiang Tonglong prescription hollow suppository on rats with prostate hyperplasia， and the effect of the proteins related to phosphoinositide 3-kinase/protein kinase B （PI3K/Akt） signaling pathway in the prostate， thus exploring the mechanism of Shengjiang Tonglong prescription hollow suppository in the treatment of rats with prostate hyperplasia. MethodTen SD male rats were randomly selected from 60 SD male rats to form a sham operation control group， and the rest rats were subcutaneously injected with testosterone propionate for 4 consecutive weeks after castration to induce the rat model of prostatic hyperplasia. According to the random number table method， the 50 rats were randomly divided into a model group， a finasteride group （0.45 mg·kg-1）， and three high， middle， and low-dose Shengjiang Tonglong prescription hollow suppository groups （3.98， 1.99， 0.99 g·kg-1）， with ten rats in each group. After castration for 7 d， the sham operation control group and the model group used the blank hollow suppositories， and the finasteride group and the Shengjiang Tonglong prescription hollow suppository groups used the corresponding hollow suppositories. The drugs were given to the rats by anal plugs continuously for 28 d. The rats were then killed， and the prostate tissues were separated and weighed to observe the effects of drugs on the prostate index of rats in each group. The hematoxylin-eosin （HE） staining was used for the pathological observation of the prostate tissues. The level of dihydrotestosterone （DHT） was detected by enzyme-linked immunosorbent assay （ELISA）. Western blot was used to detect the expression levels of PI3K/Akt signaling pathway protein， B-cell lymphoma-2 （Bcl-2）， cysteine aspartate-specific protease-3 （Caspase-3）， Bcl-2-associated X protein （Bax）， and αB-crystallin （CRYAB） protein in the prostate tissues. ResultAs compared with the sham operation control group， the protein expression levels of prostate index， DHT level， CRYAB， Bcl-2， PI3K， and Akt in the model group were increased， and the protein expression levels of Caspase-3 and Bax were decreased （P<0.05， P<0.01）. As compared with the model group， the prostate index in the high-dose Shengjiang Tonglong prescription hollow suppository group was decreased （P<0.05）， and the level of DHT and the protein expression levels of CRYAB， Bcl-2， PI3K， and Akt in the prostate of the Shengjiang Tonglong prescription hollow suppository groups were decreased， and the protein expression levels of Caspase-3 and Bax were increased （P<0.05， P<0.01）. ConclusionShengjiang Tonglong prescription hollow suppository decreases the expression of CRYAB protein， negatively regulates the PI3K/Akt signaling pathway， down-regulates the level of DHT and the protein expression levels of Bcl-2， PI3K， and Akt， and up-regulates the protein expression levels of Caspase-3 and Bax， thereby inhibiting cell proliferation and promoting cell apoptosis， which plays a therapeutic role in the benign prostate hyperplasia （BPH）. The high-dose Shengjiang Tonglong prescription hollow suppository significantly improves prostatic hyperplasia in rats.

Effect of Shengjiang Tonglong Prescription Hollow Suppository on Rats with Prostatic Hyperplasia Based on PI3K/Akt Signaling Pathway / 中国实验方剂学杂志

ObjectiveTo investigate the effect of Shengjiang Tonglong prescription hollow suppository on rats with prostate hyperplasia， and the effect of the proteins related to phosphoinositide 3-kinase/protein kinase B （PI3K/Akt） signaling pathway in the prostate， thus exploring the mechanism of Shengjiang Tonglong prescription hollow suppository in the treatment of rats with prostate hyperplasia. MethodTen SD male rats were randomly selected from 60 SD male rats to form a sham operation control group， and the rest rats were subcutaneously injected with testosterone propionate for 4 consecutive weeks after castration to induce the rat model of prostatic hyperplasia. According to the random number table method， the 50 rats were randomly divided into a model group， a finasteride group （0.45 mg·kg-1）， and three high， middle， and low-dose Shengjiang Tonglong prescription hollow suppository groups （3.98， 1.99， 0.99 g·kg-1）， with ten rats in each group. After castration for 7 d， the sham operation control group and the model group used the blank hollow suppositories， and the finasteride group and the Shengjiang Tonglong prescription hollow suppository groups used the corresponding hollow suppositories. The drugs were given to the rats by anal plugs continuously for 28 d. The rats were then killed， and the prostate tissues were separated and weighed to observe the effects of drugs on the prostate index of rats in each group. The hematoxylin-eosin （HE） staining was used for the pathological observation of the prostate tissues. The level of dihydrotestosterone （DHT） was detected by enzyme-linked immunosorbent assay （ELISA）. Western blot was used to detect the expression levels of PI3K/Akt signaling pathway protein， B-cell lymphoma-2 （Bcl-2）， cysteine aspartate-specific protease-3 （Caspase-3）， Bcl-2-associated X protein （Bax）， and αB-crystallin （CRYAB） protein in the prostate tissues. ResultAs compared with the sham operation control group， the protein expression levels of prostate index， DHT level， CRYAB， Bcl-2， PI3K， and Akt in the model group were increased， and the protein expression levels of Caspase-3 and Bax were decreased （P<0.05， P<0.01）. As compared with the model group， the prostate index in the high-dose Shengjiang Tonglong prescription hollow suppository group was decreased （P<0.05）， and the level of DHT and the protein expression levels of CRYAB， Bcl-2， PI3K， and Akt in the prostate of the Shengjiang Tonglong prescription hollow suppository groups were decreased， and the protein expression levels of Caspase-3 and Bax were increased （P<0.05， P<0.01）. ConclusionShengjiang Tonglong prescription hollow suppository decreases the expression of CRYAB protein， negatively regulates the PI3K/Akt signaling pathway， down-regulates the level of DHT and the protein expression levels of Bcl-2， PI3K， and Akt， and up-regulates the protein expression levels of Caspase-3 and Bax， thereby inhibiting cell proliferation and promoting cell apoptosis， which plays a therapeutic role in the benign prostate hyperplasia （BPH）. The high-dose Shengjiang Tonglong prescription hollow suppository significantly improves prostatic hyperplasia in rats.

Effect of Juantong Decoction on PI3K/Akt/mTOR Signaling Pathway Proteins in Rats with Endometriosis / 中国实验方剂学杂志

Objective: To discuss the effect of Juantong decoction on phosphoinositide 3-kinase/protein kinase B/mammalian target of rapamycin(PI3K/Akt/mTOR)signaling pathway proteins in rats with endometriosis (EMS). Method: The EMS Rat model was established by autologous endometrial transplantation. And 48 rats were randomly divided into 6 group, namely the sham operation group, the model group, the low-dose Juantong decoction group, the middle-dose Juantong decoction group, high-dose Juantong decoction group, and the PI3K pathway blocker group (LY294002). Then, the high-dose Juantong decoction group, the middle-dose Juantong decoction group and the low-dose Juantong decoction group were given different doses (42.9,14.3,4.8 g·kg-1). The pathway blocker group (LY294002) was given LY294002 (0.04 g·kg-1) through peritoneal injection weekly. The sham operation group and the model group were given saline irrigation (10 mL·kg-1). The administration lasted for 28 days. At last, the ectopic endometrial tissues in rats were observed by transmission electron microscope, the proteins of PI3K, Akt and mTOR in the tissue were detected by immunohistochemical method, and the p70 ribosomal S6 kinase (p70S6K) mRNA expression of ectopic endometrial tissue was tested by Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) method. Result: Compared with the normal group, the protein expressions of PI3K, Akt and mTOR and the mRNA expression of p70S6K of ectopic endometrium in the model group increased significantly (PPConclusion: The proteins of PI3K/Akt/mTOR signaling pathway participate in the occurrence of endometriosis. Juantong decoction can inhibit the activity of epithelial mesenchymal cells and promote apoptosis by reducing the protein expressions of PI3K, Akt and mTOR and the mRNA expression of P70S6K in endometriotic tissues of model rats, thus inhibiting the PI3K/Akt/mTOR signaling pathway in the treatment of endometriosis.

Effects of dexmedetomidine on PI3K/Akt pathway in hippocampus of propofol anesthetized neonatal rats / 临床麻醉学杂志

Objective To explore the effect of dexmedetomidine on phosphoinositide 3-kinase/protein kinase B (PI3K/Akt ) pathway in hippocampus of propofol anesthetized neonatal rats. Methods Eighty Sprague-Dawley male rats,aged 7 days,weighing 10-1 5 g,were randomly divided into 8 groups (n= 10 each):normal saline group (group N),DMSO group (group D),intralipid group (group I),propofol group (group P),dexmedetomidine 25 μg/kg,50 μg/kg and 75 μg/kg +propofol 100 mg/kg groups (groups PD25 ,PD50 and PD7 5 ),LY294002 25 μg + dexmedetomidine 75μg/kg + propofol 100 mg/kg group (group LYPD).The hippocampus of rats in all groups were taken 2 h after the animals fully awake.The ultrastructure of hippocampal neurons was observed by transmission electron microscope.The pAkt-(ser473 )protein and Akt protein in the hippocampus were evaluated by Western blot analysis.Results There was no significant difference in the expression of Akt protein among the eight groups.Compared with group N,the expression of pAkt (ser473)protein was significantly down-regulated in groups P,PD25 ,PD50 ,PD7 5 and LYPD (P <0.05).Compared with group P,the expression of pAkt (ser473)protein was increased significantly in groups PD7 5 and LYPD (P <0.05).Compared with group PD7 5 ,the expression of pAkt (ser473) protein was significantly down-regulated in group LYPD (P <0.05 ).The structure of hippocampal neurons was normal in groups N,I and D.Nuclear nuclei swelling,chromatin decreasing and mito-chondrion vacuolar degeneration were observed in group P while improved gradually with dexmedeto-midine in a dose-dependent manner in groups PD25 ,PD50 and PD7 5 .Neurons karyopyknosis,partial dissolution of nuclear membrane,chromatin condensation,mitochondria vacuolar degeneration were observed in group LYPD.Conclusion Dexmedetomidine pretreatment provides neuroprotection against propofol-induced hippocampal destruction by preserving PI3K/Akt pathway activity in the de-veloping brains.

B-cell activating factor involved in the pathogenesis of lupus nephritis through regulating phosphoino-sitide 3-kinase/protein kinase B/mammalian target of rapamycin signaling / 中华风湿病学杂志

Objective To investigate whether B-cell activating factor (BAFF) involved in the patho-genesis of lupus nephritis (LN) by regulating phosphoinositide 3-kinase/protein kinase B/mammalian target of rapamycin (PI3K/Akt/mTOR) signaling. Methods Twenty-eight lupus nephritis patients and 20 controls were included in this study. The clinical data were collected. BAFF levels in plasma were measured by ELISA, and the relationship between systemic lupus erythematosus disease activity index (SLEDAI) and BAFF were analyzed. The mRNA and protein levels of BAFF, phosphorylated-PI3K (p-PI3K), phosphorylated-Akt (p-Akt), phosphorylated-mTOR (p-mTOR) and Bcl-2 in kidney tissues were measured using real-time polymerase chain reaction (RT-PCR) and Western blotting. Data were analyzed using Mann-Whitney U test and Spearman correlation analysis. Results ①Plasma BAFF levels were significantly higher in LN patients [(580 ±45) ng/L] compared with controls [(208 ±30) ng/L](Z=-5.856, P<0.01), and significant positive correlation was found between plasma BAFF levels with SLEDAI (r=0.723, P<0.01). ② Plasma BAFF level in LN patients was positively correlated with 24 h UP and anti-dsDNA titers (r=0.381, 0.461, P<0.05). The protein level of BAFF in kidney tissues was positively correlated with 24 h UP and anti-dsDNA titer (r=0.469, 0.489, P<0.05).③The mRNA levels of BAFF, p-PI3K, p-Akt, p-mTOR and Bcl-2 in kidney tissues were increased in patients compared to controls[5.8±1.8 vs 2.1±0.7, Z=-4.915, P<0.01;6.7±0.9 vs 1.71±0.53, Z=-5.857, P<0.01;5.6±0.9 vs 1.8 ±0.5, Z=-5.751, P<0.01; 5.6 ±1.4 vs 1.6 ±0.4, Z=-5.291, P<0.01; 2.11 ±0.36 vs 1.33 ±0.22, Z=-4.844, P<0.01].④The protein levels of BAFF, p-PI3K, p-Akt, p-mTOR and Bcl-2 in kidney tissues were increased in patients compared to controls [0.72±0.19 vs 0.31±0.05, Z=-4.747, P<0.01;0.73±0.11 vs 0.33±0.09, Z=-5.834, P<0.01;0.77±0.06 vs 0.22±0.07, Z=-5.855, P<0.01;1.18±0.27 vs 0.47±0.13, Z=-5.416, P<0.01;2.08±0.37 vs 1.32±0.18, Z=-4.998, P<0.01]. Conclusion The findings of this study indicate that BAFF may participate in the pathogenesis of LN by regulating PI3K/Akt/mTOR signaling.

Effects of Stromal Cell-Derived-Factor-1 on Endothelial Progenitor Cells of Peripheral Blood and Their Relationship with PI3K/AKT Signal Transduction Pathway in Patients with Diabetes / 天津医药

Objective To observe the effects of stromal cell-derived-factor-1(SDF-1) on the function of endotheli?al progenitor cells(EPCs)of peripheral blood in patients with diabetes, and to discuss the effects of PI3K/AKT signaling path?way on the role of SDF-1 in EPCs. Methods The peripheral blood samples (30 mL) were collected in 10 diabetes patients (DM group) and 10 healthy controls (HC group). (1) The 100μg/L SDF-1 was added in intervention group. EGM-2MV was added in non-intervention group. The Boyden chamber and in vitro angiogenesis kit were used to analyze the migration and in vitro angiogenesis of EPCs. (2) Cultured EPCs were divided into blank control group, 1μg/L SDF-1 group, 10μg/L SDF-1 group, 100μg/L SDF-1 group, pure AMD3100 group and 100μg/L SDF-1+AMD3100 group. AKT protein expression lev?els of endothelial progenitor cells were detected by Western blot assay in each group. Results (1) Without intervention with SDF-1, EPCs’migration and angiogenesis ability were lower in DM group than those in HC group. After intervention with SDF-1, the migration and angiogenesis ability were enhanced in two groups, but the increased level was higher in DM group than that of HC group. (2) Under the same concentration, AKT protein expression level was significantly lower in DM group than that in HC group (P<0.01). AKT protein expression levels were increased with the increased levels of SDF-1 in DM group and HC group (P<0.05). AKT protein expression was significantly lower in 100μg/L SDF-1+AMD3100 group than that of 100μg/L SDF-1 group (P<0.05). Conclusion SDF-1 can increase the chemotactic migration and angiogenesis ability of EPCs in peripheral blood, especially for patients with diabetes. The effects of SDF-1 on EPCs were related to the PI3K/AKT signaling pathway.