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Pharmacological, Temperature, and Electrogram Studies on the Posterior Lymph Heart of the BullfrogOberndorfer, Carol E. 05 1900 (has links)
In view of the discrepancies and conflicts produced by previous studies on amphibian lymph hearts, a study was initiated to reinvestigate the pharmacological, temperature, and electrical aspects of lymph heart physiology. Bullfrogs were chosen as the experimental animal, All lymph heart responses to experimentation were physiographically recorded as myograms and electrograms.
The results are in agreement with previous studies on some aspects and in conflict on others. From the results obtained, lymph heart muscle appears to possess both skeletal and cardiac muscle properties as evidenced by drug responses and reactions to temperature. The precise components of the electrogram remain unclear.
It is suggested that further investigation should be made to better determine the true nature of lymph hearts.
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Molecular cloning and characterization of a cardiac and skeletal muscle LIM domain protein family (FHL). / CUHK electronic theses & dissertations collectionJanuary 1999 (has links)
Simon, Ming-yuen Lee. / Thesis (Ph.D.)--Chinese University of Hong Kong, 1999. / Includes bibliographical references (p. 239-257). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Mode of access: World Wide Web. / Abstracts in English and Chinese.
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Mechanoelectric feedback in the mammalian heart.Kelly, Douglas Robert January 2008 (has links)
Stretch of cardiac muscle is known to activate various physiological processes that result in changes to cardiac function, contractility and electrophysiology. To date, however, the precise relationship between mechanical stretch and changes in the electrophysiology of the heart remain unclear. This relationship, termed mechanoelectric feedback (MEF), is thought to underlie many cardiac arrhythmias associated with pathological conditions. These electrophysiological changes are observed not only in the whole heart, but also at the single cardiomyocyte level, and can be explained by the presence of stretch-activated ion channels (SACs). Most investigations of the actions of stretch have concentrated on these sacrolemmal ionic currents thought responsible for the proposed MEF-induced changes in contractility. While these studies have provided some useful insight into possible mechanisms, the inappropriate use of solutions and non-physiological degrees of stretch, may have caused somewhat misleading results. Currently, little is known about the involvement or contribution of non-selective or K+ selective SACs to the normal cardiac cycle. Here, I investigate the concept that stretch-induced changes in cardiac electrophysiology (MEF) are important in normal cardiac cycle and demonstrate the effects of stretch on the Frank-Starling mechanism (stretch induced increases in cardiac contractility) while pharmacologically manipulating stretch-activated ion currents. Experiments were conducted using a number of agents known to influence stretch-activated channels either in a positive or antagonistic manner. Results proved somewhat negative toward MEF theory with only substantial or pathological levels of stretch being able to elicit any electrophysiological change in the heart. Furthermore, where electrophysiological changes were associated with pathological stretch they were not consistently modulated by stretch-activated ion channel activators or blockers. Of equal importance was the observation that smaller levels of myocardial stretch associated with positive changes in contractility via the Frank-Starling mechanism were not associated with any electrophysiological changes in the Langendorff perfused heart (as observed by monophasic action potentials) nor in isolated muscle preparations (as observed through transcellular membrane potential recordings). As such, the present research undertaken in this thesis confirms an absence of electrophysiological changes with stretch except under extreme conditions suggesting that MEF is not a robust and necessarily repeatable phenomenon in the mammalian heart. / http://proxy.library.adelaide.edu.au/login?url= http://library.adelaide.edu.au/cgi-bin/Pwebrecon.cgi?BBID=1320476 / Thesis (Ph.D.) - University of Adelaide, School of Molecular and Biomedical Science, 2008
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Genetic and environmental influences on heart rate and cardiac-related autonomic activity in five-month-old twinsDubreuil, Etienne January 2002 (has links)
The first chapter of this thesis consists in a critical review of the literature on cardiac and cardiac-related autonomic activity, and infant development. Empirical findings are presented on mechanisms of interaction between heart rate and the autonomic nervous system; their relationship with infant development; their genetic and environmental influences; their gender effects; and related quantification issues. / This is followed by a study of 322 5-month-old twin pairs that investigated the genetic and environmental influences on sleeping heart rate and cardiac-related autonomic activity, as indexed by spectral analysis of heart rate variability and response to postural change. The postural change elicited only minor changes in cardiovascular activity, perhaps due to immaturity of the baroreflex. As a result, analyses focused on supine cardiovascular activity. Multivariate genetic modeling indicated that individual differences in sleeping HR and high frequency HR variability were determined by unique environmental and distinct additive genetic factors. These variables, along with low frequency HR variability, were also affected by overlapping familial environmental influences. Familial influences on individual differences in high frequency HR variability were more pronounced for baby girls than boys. Estimates of relative low and high frequency HR variability were determined by common (familial) and unique environmental factors; familial influences on these estimates of HR variability did not overlap with familial influences on sleeping HR. / A second study using the same twin sample is then presented. Its objectives were to investigate the indices of genetic and environmental etiology of individual differences in five month-old twins' HR reactivity and to evaluate the possible overlap, if any, between the familial influences on HR in states of sleep and reactivity. Multivariate genetic modeling showed that the total variance of individual differences was decomposed in the following manner: Sleeping and awake HR were under the influence of shared additive genetic factors; sleeping HR also shared common environmental influences with the absolute power spectrum values; and absolute high frequency power additionally had phenotype-specific additive genetic factors influencing its expression. Relative power spectrum values were under the influence of phenotype-specific common environmental factors. / Overall, these results suggest the presence of important familial (genetic and environmental) influences on heart rate and cardiac-related autonomic activity at five months of age: There is an absence of overlap of these (familial) additive genetic influences but the presence of a partial overlap of the (familial) environmental influences.
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Preliminary Characterization of Mitochondrial ATP-sensitive Potassium Channel (MitoKATP) Activity in Mouse Heart MitochondriaAachi, Venkat Raghav 01 March 2009 (has links)
Myocardial ischemia, infarction, heart failure and arrhythmias are the manifestations of coronary artery disease. Reduction of ischemic damage is a major concern of cardiovascular biology research. As per recent studies, the mitochondrial ATP-sensitive potassium channel (mitoKATP) opening is believed to play key role in the physiology of cardioprotection, protection against ischemia-reperfusion injury or apoptosis. However, the structural information of mitoKATP is not precisely known. Elucidating the structural integrity and functioning of the mitoKATP is therefore a major goal of cardiovascular biology research. The known structure and function of the cell ATP-sensitive potassium channel (cellKATP) is functional in interpreting the structural and functional properties of mitoKATP.
The primary goal of my research was to characterize the activity of mitoKATP in the isolated mitochondria from the control mouse heart. The mitoKATP activity, if preliminarily characterized in the control strains through the light scattering technique, then the structure of the channel could possibly be established and analyzed by means of the transgenic model and with the help of immunological techniques such as western blotting and immunoflorescence.
With this experimental model it was possible to demonstrate that the mitoKATP activity in control mouse heart mitochondria is activated by potassium channel openers (KCOs) such as diazoxide and cromakalim and activators of mitoKATP such as PMA (phorbol12 myristate-13-acetate), and inhibited by KATP inhibitors such as glibenc1amide and 5-hydroxydecanoate (5 HD).
It was evident that the KATP activity in mouse heart mitochondria was comparable to that exhibited by the rat heart mitochondria. The various selective and non-selective activators and inhibitors of the channel elicited their activity at a similar concentration used for the rat heart mitochondria. The results were reproducible in five independent experiments for each combination, further reinforcing the significance of existing channel activity in the mouse heart mitochondria.
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Effect of adenosine and lidocaine on cardiac functional and metabolic recovery after global ischemia and reperfusionVos, Lynette C. 01 January 1994 (has links)
This study investigated if exogenous adenosine (ADO) improves recovery of cardiac function during repetfusion (RPF) after global ischemia (ISC), and if lidocaine is required for. ADO-mediated cardioprotection during reperfusion. Isolated rabbit hearts, retrogradely perfused with erythrocyte-enriched Krebs-Henseleit buffer at constant left ventricular (LV) volume and physiologic flow rates, were subjected to 20 min. of global no-flow ischemia, and reperfused at the same rate as before ischemia. Hearts received one of the following treatments: 1) control (CON; no drug treatment), 2) adenosine (ADO; 200J.1M before and after ISC), or 3) adenosine+lidocaine (NL; 200 JlM ADO before and after ISC, 1 J.Lg/ml/min LIDO during first 20 min. of RPF). Myocardial function (e.g., using developed LV pressure, DP) declined as expected during no-flow ischemia and gradually returned during reperfusion. Functional recovery in ADO and NL groups were significantly improved from CON during early RPF (p<0.05 at 2 min RPF), but not at later RPF times(> 10 min). Differences did not exist between ADO and NL groups at any RPF time except at 10 min. RPF. Additionally, myocardial ATP content was measured before ischemia, after ischemia, and after 10 and 30 min of reperfusion. ATP content decreased significantly during ischemia; ADO hearts showed a increased repletion (85% of pre-ischemia level) of ATP at 30 min. of reperfusionas compared to CON (60%). These data suggest that ADO alone improves cardiac functional recovery during early repetfusion; LIDO does not appear to be required for ADO-mediated cardioprotection. ADO and LIDO do not improve cardiac function, however, ADO appears to improve myocardial ATP repletion at later RPF times in this blood-perfused rabbit model of global myocardial ISC/RPF.
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Genetic and environmental influences on heart rate and cardiac-related autonomic activity in five-month-old twinsDubreuil, Etienne January 2002 (has links)
No description available.
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Signalling pathways involved in insulin cardioprotection : are they comparable in normoxic perfused isolated rat heart vs. ischaemia/reperfusion model?Manga-Manguiya, Edith Sylvie 12 1900 (has links)
Thesis (MSc)--University of Stellenbosch, 2006. / ENGLISH ABSTRACT: Introduction: It is well documented that insulin offers cardioprotection
against the consequences of ischaemia/reperfusion injury. Insulin-induced
improvements in cardiac functions are widely investigated in models of
ischaemia and reperfusion. It has been shown that many signalling pathways
may be involved in the cardioprotection properties of insulin under those
conditions. These pathways include PI3-K, PKB/Akt, p70S6k, ERK and many
others. However, little data exists on the effects of insulin on the heart under
normoxic condition. Some evidence has been presented that insulin has a
positive inotropic effect on the normoxic perfused rat heart, but no precise
cellular mechanism has been investigated or described in this regard. We
believe that an investigation into the effects of insulin on cardiac function and
pathways involved under normoxic conditions may help us to better
understand the mechanisms of insulin-induced cardioprotection. Aims: To
determine a suitable dose of insulin at which a positive inotropic response
could be detectable under normoxic conditions, to investigate the possible
mechanisms involved in insulin-induced increases in contractility with specific
reference to the vasculature and the coronary flow and to investigate a
possible involvement of PI3-K and its downstream effectors on the insulin
effects on cardiac functions under normoxic conditions. Materials and
methods: Isolated rat hearts were perfused retrogradely using the
Langendorff technique. After 10 minutes of stabilization hearts were perfused
for 30 minutes either with standard perfusion solution i.e. Krebs-Henseleit
buffer + glucose gassed with 95%O2, 5%CO2 (control hearts), or with
standard perfusion solution plus insulin alone or insulin together with the nitric oxide synthase inhibitor L-NAME or the PI3-K inhibitor wortmannin. Left
ventricular developed pressure (LVDevP), heart rate (HR) and coronary flow
(CF) as well as phosphorylated PI3-K and PKB/Akt in heart were measured.
Results: Administration of insulin alone at physiological concentrations
showed improved cardiac function compared to hearts in the control group.
Hearts that received insulin+L-NAME showed a significant decrease in
function compared to the control hearts and the hearts that received insulin
alone (p<0.05). Phosphorylated PKB/Akt (Thr308) was increased in hearts
that received insulin alone and insulin+L-NAME compared to the control
hearts. Phosphorylated PI3-K tended to be higher in hearts where insulin was
administered alone compared to the hearts that received insulin+L-NAME or
insulin+wortmannin. Conclusion: This study confirmed that physiological
concentrations of insulin exert positive inotropic effects on cardiac function in
normoxic perfused rat hearts as seen with the improved LVDevP. Inhibition of
PI3-K by wortmannin induced a decrease in phosphorylated PKB/Akt in
hearts that received insulin+wortmannin and administration of L-NAME
impaired the beneficial effects of insulin on cardiac functions. Therefore these
results may indicate that nitric oxide may have a role in the positive effect of
insulin on cardiac function in the healthy heart perfused under normoxic
conditions. L-NAME as well as wortmannin reversed the positive inotropic
effects of insulin. Both inhibitors also unmasked effects of insulin via nitric
oxide and PI3-K on heart rate and coronary flow. / AFRIKAANSE OPSOMMING: Inleiding: Dit is welbekend dat toediening van insulien die hart beskerm
teen ischemie/reperfusie-beserings, wat lei tot verbeterde hartfunksie.
Hierdie effek word wyd ondersoek in modelle van ischemie en reperfusie. Dit
is bewys dat ‘n verskeidenheid seintransduksie paaie, insluitend PI3-K,
PKB/Akt, p70S6k en ERK, betrokke is by hierdie beskermende effek van
insulien op die hart. Baie min data is egter beskikbaar rakende die effek van
insulien tydens normoksiese toestande. Alhoewel dit bekend is dat insulien ’n
inotropiese effek op die normale geperfuseerde hart het, is die presiese
sellulêre meganismes wat dit bewerkstellig nog nie nagevors nie. Om dus ‘n
beter begrip van hierdie meganismes te verkry is dit dus noodsaaklik om die
effekte van insulien onder normoksiese perfusie toestande na te vors.
Doelstellings: Om ‘n geskikte dosis, waarby insulien sy positiewe
inotropiese effek onder normale toestande het, vas te stel, om die moontlike
meganismes betrokke by insulien-geïnduseerde verbetering in
hartsametrekbaarheid te bestudeer, met spesifieke verwysing na die
bloedvoorsiening en koronêre vloei, en om die moontlike betrokkenheid van
die PI3-K pad en sy teiken effektore onder normale suurstof-toestande te
ondersoek. Materiaal en metodes: Geïsoleerde rotharte is geperfuseer
deur gebruik te maak van die Langendorff tegniek. Na ‘n stabilisasie periode
van 10 minute is rotharte blootgestel aan 30 minute perfusie met een van vier
oplossings: ‘n standaard perfusie oplossing (Krebs-Henseleit buffer met
glukose onder spesifieke gaskondisies van 95% O2, 5% CO2 – kontrole
harte); standaard perfusie oplossing en insulien; standaard perfusie oplossing met insulien en die stikstofoksied sintase inhibitor L-NAME, of
standaard perfusie oplossing, met insulien en die PI3-K inhibitor wortmannin.
Met verloop van die perfusie protokol, is ontwikkelde linker ventrikulêre druk
(LVDevP), harttempo (HR) en koronêre vloei (CF), sowel as PI3-K en
PKB/Akt fosforilasie, gemeet. Resultate: Toediening van insulien teen
fisiologiese konsentrasies het ‘n verbeterde hartfunksie tot gevolg, in
vergelyking met harte in die kontrole groep. In teenstelling hiermee het harte
wat insulien+L-NAME ontvang het ‘n betekenisvolle verlaagde funksie getoon
in vergelyking met die kontrole harte en harte wat slegs insulien ontvang het
(p<0.05). Harte wat slegs insulien, of insulien+L-NAME ontvang het, het ‘n
verhoging in gefosforileerde PKB/Akt (Thr308) getoon in vergelyking met
kontrole harte. Gefosforileerde PI3-K het ook geneig om hoër te wees in
harte wat insulien+L-NAME of insulien+wortmannin ontvang het, as in harte
wat slegs insulien ontvang het. Gevolgtrekking: Hierdie studie bewys dat
fisiologiese konsentrasies van insulien, onder normale suurstof-toestande, ‘n
positiewe inotropiese effek op hartfunksie uitoefen, soos gesien in die
verbeterde LVDevP. Wortmannin-geïnduseerde inhibering van die PI3-K pad
het ‘n verlaagde PKB/Akt fosforilasie tot gevolg gehad in harte wat
insulien+wortmannin ontvang het, terwyl die toediening van L-NAME die
voordelige effekte van insulien op hartfunksie onderdruk het. Hierdie
resultate dui dus aan dat stikstofoksied ‘n rolspeler is in die positiewe
inotropsiese effek van insulien op hartfunksie tydens normoksiese toestande,
aangesien beide inhibitore hierdie effek onderdruk het. Beide inhibitore het
ook die betrokkenheid van stikstofoksied en die PI3-K pad by die effek van
insulien op harttempo en koronêre vloei onthul.
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Hyperglycemia-induced activation of the hexosamine biosynthetic pathway causes myocardial cell deathRajamani, Uthra 12 1900 (has links)
Thesis (PhD (Physiological Sciences))--University of Stellenbosch, 2009. / ENGLISH ABSTRACT: OBJECTIVE – Oxidative stress increases flux through the hexosamine biosynthetic pathway
(HBP) resulting in greater O-GlcNAcylation of target proteins. Since increased oxidative stress
and HBP flux are associated with insulin resistance, we hypothesized that its activation leads to
greater O-GlcNAcylation of BAD (pro-apoptotic) and increased myocardial apoptosis.
RESEARCH DESIGN AND METHODS – To investigate our hypothesis, we employed two
experimental models: 1) H9c2 cardiomyoblasts exposed to high glucose (33 mM glucose) ± HBP
modulators ± antioxidant treatment vs. matched controls (5.5 mM glucose); and 2) a rat model of
high fat diet-induced insulin resistance and hyperglycemia. We evaluated apoptosis in vitro by
Hoechst nuclear staining, Annexin-V staining, caspase activity measurements and
immunoblotting while in vivo apoptosis was assessed by immunoblotting. In vitro reactive
oxygen species (ROS) levels were quantified by H2DCFDA staining (fluorescence microscopy,
flow cytometry). We determined overall and BAD O-GlcNAcylation, both by immunoblotting
and immunofluorescence microscopy. As BAD-Bcl-2 dimer formation enhances apoptosis, we
performed immunoprecipitation analysis and immunofluorescence microscopy (co-localization)
to determine BAD-cl-2 dimerization. In vivo overall O-GlcNAcylation, BAD O-GlcNAcylation
and BAD-Bcl-2 dimerization was determined by immunoprecipitation and immunoblotting.
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RESULTS – High glucose treatment of cells significantly increased the degree of apoptosis as
revealed by Hoechst nuclear staining (54 ± 9%, p<0.01 vs. 5.5 mM), Annexin-V staining (43 ±
5%), caspase activity assay (26 ± 2%) and immunoblotting. In parallel, overall OGlcNAcylation
(p<0.001 vs. 5.5 mM), BAD O-GlcNAcylation (p<0.05 vs. 5.5 mM) and ROS
levels were increased (fluorescence microscopy – p<0.05 vs. 5.5 mM; flow cytometry – p<0.001
vs. 5.5 mM). HBP inhibition using DON and antioxidant treatment (α-OHCA) attenuated these
effects while HBP activation by PUGNAc exacerbated it. Likewise, insulin resistant rat hearts
exhibited significantly higher caspase-3 (p<0.05 vs. controls), overall O-GlcNAcylation (p<0.05
vs. controls) and BAD O-GlcNAcylation levels (p<0.05 vs. 5.5 mM). BAD-Bcl-2 dimer
formation was increased in cells exposed to hyperglycemia [immunoprecipitation analysis and
co-localization] and in insulin resistant hearts.
CONCLUSIONS - Our study identified a novel pathway whereby hyperglycemia results in
greater oxidative stress, resulting in increased HBP activation and increased BAD OGlcNAcylation.
We also found greater BAD-Bcl-2 dimerization increasing myocardial
apoptosis, suggesting that this pathway may play a crucial role in the onset of the diabetic
cardiomyopathy. / AFRIKAANSE OPSOMMING: DOELWIT – Oksidatiewe stres verhoog fluks deur die heksosamien biosintetiese weg (HBW)
wat in „n groter O-GlcNAsetilering van teiken proteïene resulteer. Weens die feit dat verhoogde
oksidatiewe stres en HBW fluks verband hou met insulienweerstandigheid, hipotetiseer ons dat
die aktivering hiervan tot groter O-GlcNAsetilering van BAD (pro-aptoptoties) en verhoogde
miokardiale apoptose lei.
NAVORSINGS ONTWERP EN METODES – Om die hipotese te ondersoek het ons twee modelle
ontplooi: 1) H9c2 kardiomioblaste is blootgestel aan hoë glukose konsentrasie (33mM glucose) ±
HBW moduleerders ± antioksidant behandeling vs. gepaarde kontrole (5.5mM glucose); en 2) „n
hoë vet dieetgeïnduseerde insulienweerstandige rotmodel en hiperglukemie. Ons het apoptose in
vitro deur middel van Hoescht nukleuskleuring geëvalueer, kasapase aktiwiteit bepalings en
immunoblotting terwyl apoptose in vivo getoets is deur immunoblotting. Reaktiewe
suurstofspesie (RSS) vlakke is deur middel van H2DCFDA verkleuring (fluoresensie
mikroskopie, vloeisitometrie) bepaal. Algehele en BAD O-GlcNAsetilering is beide deur
immunoblotting en immunofluoresensie mikroskopie bepaal. BAD-Bcl-2 dimeervorming
bevorder apoptose, om BAD-cl-2 dimerisasie te bepaal is daar van immunopresipitering analise
en immunofluoresensie mikroskopie (ko-lokalisasie) gebruik gemaak. In vivo is algehele OGlcNAsetiliering,
BAD O-GlcNAsetiliering en BAD-Bcl-2 dimerisasie deur immunopresipitasie
en immunoblotting bepaal.
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RESULTE – Hoë glukose behandeling van selle het die graad van apotpose betekenisvol verhoog
soos blootgelê deur Hoechst nukleuskleuring (54 ± 9%, p<0.01 vs. 5.5 mM), Annexin-V kleuring
(43 ± 5%), kaspase aktiviteit assay (26 ± 2%) en immunoblotting. In parallel, algehele OGlcNAsetilering
(p<0.001 vs. 5.5 mM), BAD O-GlcNAsetilering (p<0.05 vs. 5.5 mM) en RSS
vlakke is verhoog (fluoresensie mikroskopie– p<0.05 vs. 5.5 mM; vloeisitometrie– p<0.001 vs.
5.5 mM). HBW inhibering deur van DON en van antioksidant behandeling gebruik te maak (α-
OHCA) het hierdie effekte verlaag terwyl HBW aktivering deur PUGNAc dit verhoog het.
Netso, het insulienweerstandige rotharte betekenisvolle hoë kaspase -3 (p<0.05 vs. kontrole),
algeheel O-GlcNAsetilering (p<0.05 vs. kontrole) en BAD O-GlcNAsetiliering vlakke (p<0.05
vs. 5.5 mM) getoon. BAD-Bcl-2 dimeervorming is verhoog in hiperglukemies blootgestelde selle
[immunopresipitering analise en ko-lokalisering] en in insulienweerstandige harte.
GEVOLGTREKKINGS – Ons studie het „n nuwe weg geïdenifiseer waar hiperglukemie in groter
oksidatiewe stres resulteer wat weer HBW aktivering verhoog en BAD O-GlcNAsetilering
verhoog het. Ons het verder bevind dat groter BAD-Bcl-2 dimerisasie miokardiale apoptose
verhoog wat voorstel dat hierdie weg „n belangrike rol in diabetiese kardiomiopatie speel.
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The mitogenic effect of radix ophiopogonis and radix astragali on neonatal primary rat cardiomyocytes and differentiated H9C2 cardiac cells.January 2003 (has links)
Law Sui-Lin. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2003. / Includes bibliographical references (leaves 99-109). / Abstracts in English and Chinese. / CONTENTS --- p.i / ABSTRACT --- p.v / 撮要 --- p.vii / ACKNOWLEDGEMENTS --- p.ix / LIST OF FIGURES & TABLES --- p.xi / ABBREVIATIONS --- p.xv / Chapter Chapter 1 --- INTRODUCTION --- p.1 / Chapter 1.1 --- The Transition of Hyperplastic to Hypertrophic Growth During Heart Development --- p.1 / Chapter 1.2 --- The Controversial Capability of Heart Regeneration --- p.3 / Chapter 1.3 --- Challenges in Treating Heart Diseases --- p.5 / Chapter 1.4 --- A New Insight Behind Traditional Chinese Medicine (TCM) for Treating Heart Diseases --- p.7 / Chapter 1.5 --- The Potential Mitogenic TCMs on Cardiomyocytes --- p.10 / Chapter 1.5.1 --- Radix Astragali --- p.11 / Chapter 1.5.2 --- Radix Ophiopogonis --- p.12 / Chapter Chapter 2 --- MATERIALS & METHODS --- p.14 / Chapter 2.1 --- Materials --- p.14 / Chapter 2.2 --- Cell Culture --- p.16 / Chapter 2.2.1 --- Primary neonatal rat cardiomyocytes cell culture --- p.16 / Chapter 2.2.1.1 --- Mayer's hemalum-eosin staining --- p.17 / Chapter 2.2.2 --- Primary rat fibroblasts cell culture --- p.18 / Chapter 2.2.3 --- H9C2 cardiac cell culture --- p.18 / Chapter 2.3 --- TCMs Preparation and Treatment --- p.19 / Chapter 2.3.1 --- Preparation of TCMs powder from aqueous extracts --- p.19 / Chapter 2.3.2 --- Preparation of culture medium with TCMs powder --- p.19 / Chapter 2.3.3 --- Pre-treatment of undifferentiated and differentiated H9C2 cardiac cells with TCMs --- p.20 / Chapter 2.3.4 --- Post-treatment of differentiated H9C2 cardiac cells with TCMs --- p.20 / Chapter 2.4 --- Assessment of DNA Synthesis and Proliferation --- p.21 / Chapter 2.4.1 --- Tritiated thymidine incorporation assay --- p.21 / Chapter 2.4.2 --- 5-Bromo-2'-deoxy-uridine (BrdU) assay --- p.22 / Chapter 2.4.3 --- Cell counting --- p.23 / Chapter 2.4.4 --- Statistical analysis --- p.23 / Chapter 2.5 --- Screening of Differentially Expressed Genes in H9C2 Cells after TCM Treatment by cDNA Microarray --- p.25 / Chapter 2.5.1 --- Total RNA extraction --- p.25 / Chapter 2.5.2 --- RNA labeling --- p.26 / Chapter 2.5.2.1 --- Synthesis of fluorescence labeled probe --- p.26 / Chapter 2.5.2.2 --- Purification of fluorescence labeled probe --- p.27 / Chapter 2.5.3 --- Microarray hybridization --- p.28 / Chapter 2.5.3.1 --- Concentration of fluorescence labeled probe --- p.28 / Chapter 2.5.3.2 --- Hybridization --- p.28 / Chapter 2.5.3.3 --- Post-hybridization treatment --- p.29 / Chapter 2.5.4 --- Data collection --- p.29 / Chapter 2.5.4.1 --- Scanning of slide --- p.29 / Chapter 2.5.4.2 --- Image processing: spots finding and quantification --- p.30 / Chapter 2.5.5 --- Data normalization and analysis --- p.30 / Chapter 2.6 --- Confirmation of Differentially Expressed Genes in H9C2 Cells after TCM Treatment by RT-PCR --- p.32 / Chapter 2.6.1 --- DNase I digestion of total RNA sample --- p.32 / Chapter 2.6.2 --- First-strand cDNA synthesis --- p.32 / Chapter 2.6.3 --- RT-PCR of the candidate genes --- p.33 / Chapter Chapter 3 --- RESULTS --- p.36 / Chapter 3.1 --- Neonatal Primary Rat Cardiomyocytes --- p.36 / Chapter 3.1.1 --- Preparation of high-purity neonatal primary rat cardiomyocytes --- p.36 / Chapter 3.1.2 --- Neonatal primary rat cardiomyocytes ceased to undergo DNA replication after 6-day in vitro culturing --- p.38 / Chapter 3.1.3 --- Both MD and HQ promoted the growth of day 1 primary rat cardiomyocytes in dose- and time-dependent manners --- p.40 / Chapter 3.1.4 --- HQ is more potent than MD in promoting the growth of day 7 primary rat cardiomyocytes --- p.43 / Chapter 3.2 --- H9C2 Cardiac cells --- p.45 / Chapter 3.2.1 --- Proliferative effect of MD and HQ on undifferentiated H9C2 cardiac cells --- p.45 / Chapter 3.2.2 --- Pre-treatment of HQ on H9C2 cardiac cells during differentiation --- p.50 / Chapter 3.2.3 --- Pre-treatment of MD and HQ on differentiated H9C2 cardiac cells --- p.52 / Chapter 3.2.4 --- Post-treatment of MD on differentiated H9C2 cardiac cells…… --- p.55 / Chapter 3.3 --- Primary Rat Fibroblasts --- p.57 / Chapter 3.3.1 --- Proliferative effect of MD and HQ on primary rat fibroblasts --- p.58 / Chapter 3.4 --- Screening of Differentially Expressed Genes in H9C2 Cells after HQ Treatment by cDNA Microarray --- p.60 / Chapter 3.4.1 --- Differentially expressed genes in undifferentiated H9C2 cardiac cells after HQ treatment --- p.60 / Chapter 3.4.2 --- Differentially expressed genes in differentiated H9C2 cardiac cells after HQ treatment --- p.66 / Chapter 3.4.3 --- Comparison of differentially expressed genes in both undifferentiated and differentiated H9C2 cardiac cells after HQ treatment --- p.72 / Chapter 3.5 --- Confirmation of Differentially Expressed Genes in H9C2 Cells after HQ Treatment by RT-PCR --- p.73 / Chapter 3.5.1 --- "Preferential up-regulation of N-G, N-G-dimethylarginine dimethylaminohydrolase mRNA expression level in undifferentiated H9C2 cardiac cells after HQ treatment " --- p.74 / Chapter 3.5.2 --- Preferential up-regulation of heme oxygenase-3 mRNA expression level in undifferentiated H9C2 cardiac cells after HQ treatment --- p.75 / Chapter 3.5.3 --- Preferential up-regulation of cyclin B mRNA expression level in differentiated H9C2 cardiac cells after HQ treatment --- p.76 / Chapter Chapter 4 --- DISCUSSION --- p.77 / Chapter 4.1 --- HQ Being a More Effective Mitogenic TCM than MD on Cardiomyocytes Exerted its Effect in Dose- and Time Dependent --- p.79 / Chapter 4.2 --- Mitogenic Effect of Both MD and HQ might Possibly Due to the Regulation of Intrinsic Factors --- p.82 / Chapter 4.3 --- HQ Rather Than MD Showed a Higher Specificity in Promoting DNA Synthesis in Cardiomyocytes --- p.83 / Chapter 4.4 --- The Differentially Expressed Genes were Supported by The Clinical Functions of HQ --- p.85 / Chapter 4.5 --- Relating the Differentially Expressed Genes with Cardiac Growth and Development --- p.87 / Chapter 4.6 --- The Hypothetic Mechanisms of Action that HQ Exerted on Cardiac Growth and Development --- p.92 / Chapter 4.7 --- Future Prospect --- p.94 / Chapter 4.7.1 --- In vivo study of HQ on the proliferation of rat cardiomyocytes from neonatal to postnatal development --- p.94 / Chapter 4.7.2 --- The study of transgenic mice carrying the target gene regulated by HQ on cardiac growth and development --- p.96 / Chapter 4.7.3 --- The determination of active component of HQ on cardiac growth and development --- p.97 / REFERENCES --- p.99 / APPENDIX --- p.110
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