OBJECTIVE: To investigate the protective efficacy of electroacupuncture(EA) pretreatment at Neiguan(PC6) on myocardial ischemia-reperfusion(I/R) in rats.METHODS: Fifty rats were randomly divided into five groups(n = 1...OBJECTIVE: To investigate the protective efficacy of electroacupuncture(EA) pretreatment at Neiguan(PC6) on myocardial ischemia-reperfusion(I/R) in rats.METHODS: Fifty rats were randomly divided into five groups(n = 10): sham operation group, model group(underwent in vivo myocardial I/R), EA pretreatment group [EA at Neiguan(PC 6) 1 week before I/R], wortmannin group(1 week before I/R, the PI3K inhibitor, wortmannin, was injected), EA pretreatment + wortmannin group(both pretreatments were performed simultaneously). After establishing the I/R model, 2,3,5-triphenyltetrazolium chloride(TTC) staining was used to analyze the weight and area of the myocardial infarction tissue.The biosignal and pressure test system was used to determine the left ventricular systolic mean pressure(LVSP), left ventricular end-diastolic pressure(LVEDP), fractional shortening(FS), and ejection fraction(EF). Ultraviolet spectrophotometry was used to determine the expression of creatine kinase(CK)-MB, inducible nitric oxide synthase(i NOS), and total antioxidant capacity(T-AOC) in the serum. The expression of autophagy-related protein 13(ATG13), mammalian target of rapamycin(m TOR), and phosphatidylinositol 3-kinase(PI3K) in cardiac muscle cells was determined by immunofluorescence. Hematoxylin and eosin staining was used to observe autophagy-related pathological changes in rat cardiomyocytes, and ultrastructural changes of cardiomyocytes were examined by transmission electron microscopy.RESULTS: In this study, the infarction size and tissue weight of the EA pretreatment group were decreased compared with the model group(P <0.0001). Furthermore, compared with the model group, the LVEDP values of the EA pretreatment group were significantly reduced(P = 0.0091), and the LVSP, FS, and EF values were slightly increased (P = 0.0007, 0.0020, 0.0031). EA pretreatment also significantly decreased the expression of CK-MB and i NOS, while it increased the expression of T-AOC in the serum of rats with I/R injury(P <0.0001). Furthermore, EA pretreatment slightly widened the myocardial fiber space, reduced necrosis and myocardial cell swelling and maintained the nucleus and mitochondria structure intact.CONCLUSION: EA pretreatment promoted autophagy flux and alleviated myocardial I/R injury through the PI3K-Akt-m TOR pathway.展开更多
OBJECTIVE:To investigate the efficacy of self-made Gengnian decoction on expressions of phosphatidylinositol 3-kinase(PI3 K),protein kinase B(Akt)and mammalian target of rapamycin(m TOR)in ovarian tissues of perimenop...OBJECTIVE:To investigate the efficacy of self-made Gengnian decoction on expressions of phosphatidylinositol 3-kinase(PI3 K),protein kinase B(Akt)and mammalian target of rapamycin(m TOR)in ovarian tissues of perimenopausal rats.They were identified with symptom pattern of kidney-Yang deficiency in terms of Traditional Chinese Medicine.METHODS:Female Sprague-Dawley rats aged10-12 months were selected.Estrous cycle was observed by vaginal smears of keratinocytes to screen the perimenopausal model rats.The chosen rats were randomly divided into five groups,including perimenopausal model of kidney-Yang deficiency group(24 rats),self-made Gengnian decoction of high-dose group(24 rats),self-made Gengnian decoction of middle-dose group(24 rats),self-made Gengnian decoction of low dose group(24 rats)and tibolone control group(24 rats).In addition,rats aged 4-6 months were selected as young control group.The perimenopausal model rats of kidney-Yang deficiency were prepared by alternative intramuscular injection of hydrocortisone 5 mg·kg^-1·d^-1The successfully prepared models in self-made Gengnian decoction of high-dose,middle-dose and low-dose groups and tibolone control group were given self-made Gengnian decoction 26.4,13.2 and 6.6 mg·kg^-1·d^-1,and tibolone tablets solvent 0.22 mg·kg^-1·d^-1,respectively,through intragastric administration.Models group and young control group were given the same dose of normal saline,1 time a day for 15 consecutive days.24 h after the last administration,blood and ovarian tissues were collected after anesthesia with 20%ethyl carbamate.The follicles of different levels in ovarian tissue were observed and counted by histopathological hematoxylin-eosin staining.Enzyme linked immunosorbent assay was applied to test insulin-like growth factor-1(IGF-1)level in the serum of experimental rats.The expression levels of PI3 K,phosphorylated-Akt(p-Akt)and phosphorylated-m TOR(p-m TOR)m RNA in ovarian tissue were detected by quantitative real-time polymerase chain reaction.RESULTS:The total follicle counts of perimenopausal model rats with kidney-Yang deficiency were significantly reduced,and the number of follicles(mainly increased in preantral follicles and antral follicles)in perimenopausal model rats with kidney-Yang deficiency was significantly increased after intervention of high and middle doses of Gengnian decoction and tibolone(P<0.05).Compared with normal rats in young control group,the levels of IGF-1 in serum of perimenopausal rats with kidney-Yang deficiency were significantly decreased(P<0.01),and those intervened by high dose of Gengnian decoction and tibolone were significantly up-regulated.The relative expression levels of PI3 K,p-Akt,p-m TOR m RNA in ovarian tissues of perimenopausal rats with kidney-Yang deficiency were significantly lower than those of young rats(P<0.01),and those intervened by high dose of Gengnian decoction and tibolone were significantly up-regulated(P<0.05).CONCLUSION:Self-made Gengnian decoction can increase the levels of IGF-1,PI3 K,Akt and m TOR m RNA expression in serum.展开更多
OBJECTIVE:To explore the protective mechanisms of the Traditional Chinese Medicine Bushenhuoxue(BSHX)in a rat model of vascular dementia(VD).METHODS:A rat model of VD was developed using bilateral common carotid arter...OBJECTIVE:To explore the protective mechanisms of the Traditional Chinese Medicine Bushenhuoxue(BSHX)in a rat model of vascular dementia(VD).METHODS:A rat model of VD was developed using bilateral common carotid artery occlusion(BCCAO).Rats were administered BSHX(10.14 or 5.07 g/kg),nimodipine(11.06 mg/kg;positive control),or saline(control)by gavage daily for 30 d post-surgery.Learning and memory abilities were assessed using the Morris water maze.Morphological changes in the hippocampus were observed using light microscopy(hematoxylin and eosin staining)and transmission electron microscopy(TEM).The m RNA and protein expression levels of brain-derived neurotrophic factor(BDNF),tyrosine receptor kinase B(Trk B),phosphatidyl inositol 3-kinase(PI3 K),serine/threonine kinase(AKT),and c AMP response element binding protein(CREB)were measured by real-time polymerase chain reaction(RT-PCR)and Western blot,respectively.RESULTS:Compared with the sham group,rats with BCCAO exhibited impaired learning and memory abilities(Morris water maze)and showed abnormalities in neuronal morphology(light microscopy)and ultrastructure(TEM)in the hippocampus.They also had decreased m RNA and protein expressions of BDNF,Trk B,PI3 K,AKT,and CREB in hippocampal tissue(all P<0.05).In rats with BCCAO,administration of BSHX attenuated deficits in learning and memory,improved the morphology and ultrastructure of hippocampal neurons,and enhanced m RNA and protein expression levels of BDNF,Trk B,PI3 K,AKT,and CREB(all P<0.05).CONCLUSION:BSHX may protect hippocampal neurons and improve learning and memory abilities,at least in part via the activation of BDNF/Trk B/PI3 K/AKT/CREB signaling.展开更多
基金Supported by Hubei Provincial Department of Education Science and Technology Research Program Youth Talent Project:Mechanism of Electroacupuncture Preconditioning on Myocardial Ischemia-Reperfusion Injury Cell Apoptosis Through MiR-133-3p-Mediated Rho A/p38MAPK Pathway(No.Q20192012)。
文摘OBJECTIVE: To investigate the protective efficacy of electroacupuncture(EA) pretreatment at Neiguan(PC6) on myocardial ischemia-reperfusion(I/R) in rats.METHODS: Fifty rats were randomly divided into five groups(n = 10): sham operation group, model group(underwent in vivo myocardial I/R), EA pretreatment group [EA at Neiguan(PC 6) 1 week before I/R], wortmannin group(1 week before I/R, the PI3K inhibitor, wortmannin, was injected), EA pretreatment + wortmannin group(both pretreatments were performed simultaneously). After establishing the I/R model, 2,3,5-triphenyltetrazolium chloride(TTC) staining was used to analyze the weight and area of the myocardial infarction tissue.The biosignal and pressure test system was used to determine the left ventricular systolic mean pressure(LVSP), left ventricular end-diastolic pressure(LVEDP), fractional shortening(FS), and ejection fraction(EF). Ultraviolet spectrophotometry was used to determine the expression of creatine kinase(CK)-MB, inducible nitric oxide synthase(i NOS), and total antioxidant capacity(T-AOC) in the serum. The expression of autophagy-related protein 13(ATG13), mammalian target of rapamycin(m TOR), and phosphatidylinositol 3-kinase(PI3K) in cardiac muscle cells was determined by immunofluorescence. Hematoxylin and eosin staining was used to observe autophagy-related pathological changes in rat cardiomyocytes, and ultrastructural changes of cardiomyocytes were examined by transmission electron microscopy.RESULTS: In this study, the infarction size and tissue weight of the EA pretreatment group were decreased compared with the model group(P <0.0001). Furthermore, compared with the model group, the LVEDP values of the EA pretreatment group were significantly reduced(P = 0.0091), and the LVSP, FS, and EF values were slightly increased (P = 0.0007, 0.0020, 0.0031). EA pretreatment also significantly decreased the expression of CK-MB and i NOS, while it increased the expression of T-AOC in the serum of rats with I/R injury(P <0.0001). Furthermore, EA pretreatment slightly widened the myocardial fiber space, reduced necrosis and myocardial cell swelling and maintained the nucleus and mitochondria structure intact.CONCLUSION: EA pretreatment promoted autophagy flux and alleviated myocardial I/R injury through the PI3K-Akt-m TOR pathway.
基金Supported by National Natural Science Foundation of China(No.81560786),project funds from"Study on the Protective Mechanism of Tonifying Kidney,Warming Yang,Dispelling Wind and Cold Method Based on PI3K/Akt/m TOR Signaling Pathway on Ovarian Function in Rats with Kidney-Yang Deficiency"Project of Guizhou Provincial Administration of Traditional Chinese Medicine,"Regulation of IGF-1 Expression on P13K/Akt/m TOR Signaling Pathway in Ovarian Granulosa Cells of Kidney-Yang Deficiency Rats and Intervention of Gengnian Decoction"(No.QZYY-2016-021)issued by Guizhou Provincial Administration of Traditional Chinese MedicineGuizhou Provincial Joint fund for Science and Technology"Study on the Protective Mechanism of Tonifying Kidney,Warming Yang,Dispelling Wind and Cold Method based on PI3K/Akt/m TOR Signaling Pathway on Ovarian Function in Rats with Kidney-Yang Deficiency"(No.Guizhou[2015]7225)
文摘OBJECTIVE:To investigate the efficacy of self-made Gengnian decoction on expressions of phosphatidylinositol 3-kinase(PI3 K),protein kinase B(Akt)and mammalian target of rapamycin(m TOR)in ovarian tissues of perimenopausal rats.They were identified with symptom pattern of kidney-Yang deficiency in terms of Traditional Chinese Medicine.METHODS:Female Sprague-Dawley rats aged10-12 months were selected.Estrous cycle was observed by vaginal smears of keratinocytes to screen the perimenopausal model rats.The chosen rats were randomly divided into five groups,including perimenopausal model of kidney-Yang deficiency group(24 rats),self-made Gengnian decoction of high-dose group(24 rats),self-made Gengnian decoction of middle-dose group(24 rats),self-made Gengnian decoction of low dose group(24 rats)and tibolone control group(24 rats).In addition,rats aged 4-6 months were selected as young control group.The perimenopausal model rats of kidney-Yang deficiency were prepared by alternative intramuscular injection of hydrocortisone 5 mg·kg^-1·d^-1The successfully prepared models in self-made Gengnian decoction of high-dose,middle-dose and low-dose groups and tibolone control group were given self-made Gengnian decoction 26.4,13.2 and 6.6 mg·kg^-1·d^-1,and tibolone tablets solvent 0.22 mg·kg^-1·d^-1,respectively,through intragastric administration.Models group and young control group were given the same dose of normal saline,1 time a day for 15 consecutive days.24 h after the last administration,blood and ovarian tissues were collected after anesthesia with 20%ethyl carbamate.The follicles of different levels in ovarian tissue were observed and counted by histopathological hematoxylin-eosin staining.Enzyme linked immunosorbent assay was applied to test insulin-like growth factor-1(IGF-1)level in the serum of experimental rats.The expression levels of PI3 K,phosphorylated-Akt(p-Akt)and phosphorylated-m TOR(p-m TOR)m RNA in ovarian tissue were detected by quantitative real-time polymerase chain reaction.RESULTS:The total follicle counts of perimenopausal model rats with kidney-Yang deficiency were significantly reduced,and the number of follicles(mainly increased in preantral follicles and antral follicles)in perimenopausal model rats with kidney-Yang deficiency was significantly increased after intervention of high and middle doses of Gengnian decoction and tibolone(P<0.05).Compared with normal rats in young control group,the levels of IGF-1 in serum of perimenopausal rats with kidney-Yang deficiency were significantly decreased(P<0.01),and those intervened by high dose of Gengnian decoction and tibolone were significantly up-regulated.The relative expression levels of PI3 K,p-Akt,p-m TOR m RNA in ovarian tissues of perimenopausal rats with kidney-Yang deficiency were significantly lower than those of young rats(P<0.01),and those intervened by high dose of Gengnian decoction and tibolone were significantly up-regulated(P<0.05).CONCLUSION:Self-made Gengnian decoction can increase the levels of IGF-1,PI3 K,Akt and m TOR m RNA expression in serum.
基金Supported by Hebei Province Natural Science Fund(Protection Mechanism Research of Bushenhuoxue on Hippocampal Nerve in Rats with Vascular Dementia Based on BDNF/Trk B Signaling Pathway,No.H2015423057)Hebei Provincial Department of Education Science and Technology Research Key Funding Project(Study on the Mechanism of Regulating the Autophagy and Apoptosis of Hippocampal Neurons in VD Rats by Bushenhuoxue Prescription,No.ZD2018009)Research Capacity Enhancement Project of Hebei University of Chinese Medicine(Improvement of Hippocampal Synaptic Remodeling in Rats with Vascular Dementia by Regulating BDNF/Trk B Signaling Pathway Based on the"renal essence"theory,No.2019-11).
文摘OBJECTIVE:To explore the protective mechanisms of the Traditional Chinese Medicine Bushenhuoxue(BSHX)in a rat model of vascular dementia(VD).METHODS:A rat model of VD was developed using bilateral common carotid artery occlusion(BCCAO).Rats were administered BSHX(10.14 or 5.07 g/kg),nimodipine(11.06 mg/kg;positive control),or saline(control)by gavage daily for 30 d post-surgery.Learning and memory abilities were assessed using the Morris water maze.Morphological changes in the hippocampus were observed using light microscopy(hematoxylin and eosin staining)and transmission electron microscopy(TEM).The m RNA and protein expression levels of brain-derived neurotrophic factor(BDNF),tyrosine receptor kinase B(Trk B),phosphatidyl inositol 3-kinase(PI3 K),serine/threonine kinase(AKT),and c AMP response element binding protein(CREB)were measured by real-time polymerase chain reaction(RT-PCR)and Western blot,respectively.RESULTS:Compared with the sham group,rats with BCCAO exhibited impaired learning and memory abilities(Morris water maze)and showed abnormalities in neuronal morphology(light microscopy)and ultrastructure(TEM)in the hippocampus.They also had decreased m RNA and protein expressions of BDNF,Trk B,PI3 K,AKT,and CREB in hippocampal tissue(all P<0.05).In rats with BCCAO,administration of BSHX attenuated deficits in learning and memory,improved the morphology and ultrastructure of hippocampal neurons,and enhanced m RNA and protein expression levels of BDNF,Trk B,PI3 K,AKT,and CREB(all P<0.05).CONCLUSION:BSHX may protect hippocampal neurons and improve learning and memory abilities,at least in part via the activation of BDNF/Trk B/PI3 K/AKT/CREB signaling.