Sc12L8 Acid-Base Regulation pH & Buffers: Definition of pH -> -log[H+] -> pH = 7.4 -> a.k.a.

. 40 nM H+ Normal range -> 7.35 to 7.45 -> 35 to 45 nM H+ Plasma pH lower than 7.35 is acidosis, above 7.45 is alkalosis Plasma pH outside range of 7.1 to 7.7 may be incompatible with life -> pH affects enzyme rates Buffers -> weak acids or bases -> combine reversibly with H+ o Weak acid -> HA = H+ + Ao Weak base -> B + H+ = BH+ o Buffer effectiveness depends on conc & pK (log of dissociation constant of HA or BH+) o Main buffers are proteins His residues -> pK ~ 6 Ammonia -> pK 9.2 Phosphate -> pK 6.8 CO2 pK -> 6.1 Urate pK -> 5.8 Citrate -> pK 5.5 o Buffer cant prevent pH change in response to H+ or OH- -> only makes change smaller than without buffer H+ can enter cells readily, mainly in exchange for K+, then buffered by intracellular proteins

CO 2 /Bicarbonate Buffer System: - CO2/HCO3- system is v imp buffer in blood o CO2 + H2O = H2CO3 = H+ + HCO3o H2CO3 = H+ + HCO3- is fast o CO2 + H2O = H2CO3 slow, but catalysed by enzyme carbonic anhydrase -> present in erythrocytes & epithelia e.g. gut & kidney - Both reactions normally in equilibrium & Henderson-Hasselbalch equation applies - Allows calculation of pH, PCO2 & HCO3- conc if 2 are known o In practise, pH & PCO2 measured with blood-gas analyser & HCO3- conc calculated - Normal values o PaCO2 = 5.3 kPa o pH = 7.4 o HCO3- = 25 mM

Sc12L8 Acid-Base Regulation

Respiratory Buffering: - Ventilation inc as PaCO2 inc - If breathe faster, PCO2 dec & PO2 inc o Assumes O2 consumption & CO2 production rates are constant - Inc PaCO2 stimulates respiration -> inc CO2 excretion through lungs - Inc in H+ causes inc in PaCO2 o Due to reaction CO2 + H2O = H2CO3 = H+ + HCO3o Inc in PaCO2 will stimulate respiration & some extra CO2 expired o Final change in H+ conc smaller than otherwise would occur o Reverse applies if H+ dec - So CO2 + H2O = H2CO3 = H+ + HCO3- buffering system works with respiratory system to minimise pH changes o Mechanism may not be effect if respiratory function compromised o Cant replace kidneys in excreting acid substances e.g. NH4+, 2H+, SO42Metabolic Production of H + : - Oxidative metabolism of fats & carbohydrates produces H2O & CO2 -> excreted in lungs - CO2 production -> 12 to 20 moles per day -> sometimes called volatile acidity - Anaerobic metabolism of carbohydrates leads to production of lactate (weak acid) -> other metabolic processes lead to production of citrate, acetoacetate, urate & sulphate -> from oxidation of cysteine - Oxidation of neutral AA leads to production of NH4+ plus HCO3- Overall production of acid metabolites varies with diet -> usually equivalent to 50 to 100 mmol of H+ per day -> acid cant be excreted in lungs so has to go through kidney -> a.k.a. fixed acidity - Urine usually acidic -> pH range 5 to 7 -> acidity measured in titration with NaOH to pH 7-> value = titratable acidity - Production of acetoacetate & -hydroxybutyrate (ketone bodies) by liver inc greatly in starvation & diabetes mellitus -> inhibited by insulin Principles of H + Regulation by kidney: - Secreted into nephron by active transport in proximal & distal tubule & collecting duct - Secretion regulated according to bodys needs - Stimulated by inc PCO -> CO2 acts as source for H+ - Plasma normally contains 25 mM HCO3- -> same conc found in glomerular filtrate -> usually all reabsorbed by reaction with H+ ions which are excreted in tubule H+ + HCO3- = CO2 + H2O = H2CO3 (reabsorbed) - Excess H+ secreted in urine, buffer H2PO4- (pKa 6.8) or NH4+ (pKa 9.2) - H+ excretion req buffer -> unbuffered H+ lowers pH to below 5 -> H+ stop working
2

Sc12L8 Acid-Base Regulation

Tubular Mechanisms: - Proximal tubule: o 80% of filtered bicarbonate reabsorbed by pathways in proximal tubule -> doesnt contribute to net H+ excretion o 2 mechanisms for H+ secretion at luminal surface Na+/H+ exchange (major) ATP dependent H+ pump (minor) + o H derives from CO2 & production inhibited by carbonic anhydrase e.g. diamox 2 bicarbonate exchangers at blood surface o Glutamine made in liver -> deaminated -> yields 2-oxoglutarate & 2NH4+ o NH4+ goes into urine -> either permeable NH3 & H+ or NH4+ on Na+/H+ exchanger o 1H+ used in 2-oxoglutarate metabolism & HCO3- to plasma o Each NH4+ excreted carries 1H+ ion - Thick ascending limb of loop of Henle o Normally ~ 80% of filtered bicarbonate reabsorbed in proximal tubule & 10 to 15% more in thick ascending limb Result of H+ secretion via Na+/H+ exchanger o Some of NH4+ secreted earlier in nephron can be reabsorbed in thick ascending limb via Na+/K+/2Cl- co-transporter - Collecting duct o Intercalated type A cell (more numerous than B) H+ pumped out mainly by H+/K+-ATPase For every H+ pumped out, HCO3- made in kidney & returned to blood o Intercalated type B cell work reverse -> transfer H+ to blood -> function unknown Renal Regulation of H + & Bicarbonate Excretion: - If plasma HCO3- conc varied acutely -> behaves as if there were Tm for reabsorption such that 25 mM HCO3- conc, its completely reabsorbed - If theres excess HCO3-, like alkalosis, excreted in urine -> not true Tm, as it varies with PaCO2 - Secretion of H+ into kidney tubules req H+ to be produced in cell from CO2 using carbonic anhydrase - Acute changes of PaCO2 in dog affect max capacity for renal bicarbonate reabsorption -> affect availability of H+ -> evidence that CO2 is source of H+ ions in kidney

Sc12L8 Acid-Base Regulation

Hepatic Regulation of H + Excretion: - Liver metabolises excess AA -> yields NH4+ & HCO3- & removed via 2 pathways, according to plasma pH o Excretion of urea -> urea products via ornithine-citrulline cycle -> many intermediate steps Overall reaction: 2HCO3- + 2NH4+ -> CO(NH2)2 + CO2 + 3H2O Mechanism predominated in alkalosis Consumes HCO3- & urea (product) excreted o Excretion of glutamine -> glutamine synthase present in perivenous hepatocytes NH4+ + glutamate = glutamine o Mechanism predominates in acidosis o Not consume HCO3- -> HCO3- produced by AA metabolism returned to blood - Kidney excretes H+ as NH4+ -> availability of NH4+ totally dependent on synthesis of glutamine in liver, which is pH dependent - In isolated, perfused rat liver, rate of urea & glutamine production vary according to pH - Glutamine synthesis predominates at low pH while urea production predominates at high pH - Glutamine synthesis & breakdown enzymes also show pH dependence o Glutaminase activity predominating at high pH o Have diff locations Glutamine synthase in perivenous hepatocytes Glutaminase in periportal hepatocytes Acid-Base Disorders: - If arterial pH below 7.35 -> acidosis disorder, if above 7.45 -> alkalosis disorder - 2 forms of acidosis & alkalosis distinguished -> respiratory & metabolic o Respiratory acidosis or alkalosis caused by fault in lungs Will show up in arterial PCO2 -> elevated in acidosis & reduced in alkalosis o Metabolic acidosis or alkalosis caused by fault not in lungs Arterial PCO2 likely to be changed in opposite direction -> less than 5.3 kPa in acidosis -> because of respiratory compensation for acidosis - Normally both kidneys & lungs involved in pH regulation o If pH disturbed, can both participate in dec pH change o Processes called respiratory compensation & renal compensation

Sc12L8 Acid-Base Regulation

Respiratory Acidosis: - Caused by insufficient CO2 excretion by lungs -> caused by e.g. lung damage, respiratory muscle weakness, brainstem damage ->patient likely to be hypoxic (low PaCO2 as well) - Some of retained CO2 converted to HCO3- by reaction CO2 + H2O = H2CO3 = H+ + HCO3- -> arterial PCO2 inc, pH dec & HCO3- conc inc - Kidneys inc H+ secretion -> result in complete reabsorption of filtered HCO3- & H+ excretion at inc rates -> renal compensation for respiratory acidosis makes pH dec smaller than otherwise, but PCO2 & HCO3- conc elevated & still acidosis Respiratory Alkalosis: - Caused by hyperventilation -> caused by e.g. hypoxia -> high altitudes or dec alveolar diffusion or ventilation-perfusion mismatch or severe anaemia - Ventilation may also be stimulated voluntarily -> anxiety, pregnancy (progesterone), as side effect of salicylate treatment & other diseases - Excess CO2 lost in lungs, PCO2 falls & HCO3- conc dec as some reacts with H+ to make CO2 - Kidneys respond to fall in PCO2 with dec in H+ secretion into tubules - Now insufficient H+ to ensure reabsorption of all HCO3- in filtrate & HCO3- excreted in urine -> alkaline - Kidneys compensate for respiratory alkalosis by excreting alkaline urine - End result -> low PCO2, high pH & low HCO3- conc -> smaller changes, than otherwise Metabolic Acidosis: - Caused by: o Ingestion of acids or ammonium salts o Metabolism-producing acids -> lactate in excersise, ketoacids in starvation or diabetes mellitus o Renal failure or impairment of H+ transport o Excess loss of bicarbonate from gut -> e.g. diarrhoea - Inc in H+ causes HCO3- conc dec, PCO2 rises, due to reaction H+ + HCO3= H2CO3 = H2O + CO2 - Rise in PCO2 stimulate respiration & also stimulate H+ secretion by kidney (except in renal failure or impairment of H+ transport) - End result -> low pH (acidosis), low HCO3- conc & usually low PCO2 (could be elevated if mixed metabolic & respiratory acidosis)

Sc12L8 Acid-Base Regulation

Metabolic Alkalosis: - Caused by -> ingestion of bicarbonate, excess loss of acid from body via vomiting, high levels of aldosterone, which indirectly stimulates H+ excretion in collecting duct due to Na+ channels opening - Characterised by pH greater than 7.45 with high plasma HCO3- Inc in plasma HCO3- conc & inc in pH -> there wont be significant direct rise in PCO2 -> due to H+ conc only needs to dec by few nM - Rise in pH should inhibit respiration -> causes CO2 to retain in body - Resulting hypoxia may continue to stimulate respiration - Kidneys excrete excess bicarbonate unless prevented from doing so by high levels of aldosterone o These should help to dec the rising pH Anion Gap: - Calculated from conc of plasma electrolytes -> (Na+ + K+) (Cl- + HCO3-) - Normal range 6 to 16 mM - Anion gap inc in some types of metabolic acidosis -> can be used in diagnosis - If theres gut bicarbonate loss (e.g. diarrhoea), chloride replaces bicarbonate & anion gap normal -> same in some forms of renal tubular acidosis characterised by bicarbonate loss - If fresh acid added to body, anion gap enhanced o E.g. lactate acidosis or diabetic ketoacidosis o Acid converts bicarbonate to CO2 & no corresponding inc in chloride, so anion gap inc

Sc12L8 Acid-Base Regulation