Increased plasma volume in two models of portal hypertension in the rat: cirrhosis of the liver and partial portal vein ligation

Portal hypertension has been studied in the rat to see if it is associated to altered blood volume composition, as it has been shown in other species. Plasma volume was measured by isotope dilution using 99mTc labelled albumin in three groups of male Sprague-Dawley rats: normal rats (controls), partially ligated portal vein rats and rats with Cl4C induced cirrhosis. Plasma volume was significantly higher in rats with portal hypertension due to partially ligated portal vein and cirrhosis than in control animals. Similarly, the calculated blood volume was also significantly higher in the portal hypertensive animals than in control group. Portal hypertension in the rat, therefore, has been demonstrated to be associated to a marked hypervolemia and this finding should be taken into consideration in haemodynamic and pharmacokinetic studies in portal hypertensive rat models.     

Proinflammatory liver and antiinflammatory intestinal mediators involved in portal hypertensive rats

Proinflammatory (TNF-alpha , IL-1beta, and NO) and antiinflammatory (IL-10, CO) levels were assayed in serum, liver, and small bowel in order to verify a hypothetic inflammatory etiopathogeny of portal hypertension that could be the cause of its evolutive heterogeneity. Male Wistar rats were divided into one control group (n=11) and one group with a triple stenosing ligation of the portal vein (n=23) after 28 days of evolution. In one subgroup of portal hypertensive rats, portal pressure, collateral venous circulation, mesenteric vasculopathy, and liver and spleen weights were determined. In the remaining rats with portal hypertension TNF-alpha, IL-1beta, and IL-10 were quantified in liver and ileum by enzyme-linked immunosorbent assay. NO synthase activity was studied in liver and ileum. CO and NO were measured in portal and systemic blood by spectrophotometry and Griess reaction, respectively. Portal hypertensive rats with mayor spleen weight show hepatomegaly and mayor development of collateral circulation. Ileum release of IL-10 (0.30 +/- 0.12 versus 0.14 +/- 0.02 pmol/mg protein; P< .01) is associated with a liver production of both proinflammatory mediators (TNF-alpha: 2 +/- 0.21 versus 1.32 +/- 0.60 pmol/mg protein; P< .05, IL-1beta: 19.17 +/- 2.87 versus 5.96 +/- 1.84 pmol/mg protein; P=.005, and NO: 132.10 +/- 34.72 versus 61.05 +/- 8.30 nmol/mL; P=.005) and an antiinflammatory mediator (CO: 6.49 +/- 2.99 versus 3.03 +/- 1.59 pmol/mL; P=.005). In short-term prehepatic portal hypertension a gut-liver inflammatory loop, which could be fundamental in the regulation both of the portal pressure and of its complications, could be proposed.     

Effect of portacaval surgical anastomosis on systemic and splanchnic hemodynamics in portal hypertensive, cirrhotic rats

The effect of surgical end-to-side portacaval anastomosis (PCSA) on systemic and splanchnic circulation has been studied in cirrhotic rats with portal hypertension (CCl4-phenobarbital method) and in control animals. Hemodynamics have been measured using the microsphere technique, with a reference sample for the systemic hemodynamic measurements, and intrasplenic injection for portal systemic shunting rate measurements. Compared with controls, sham-operated (SO) cirrhotic rats showed a hyperdynamic circulation with increased cardiac output (CO) and decreased mean arterial pressure and peripheral resistances. PCSA in control rats induced only a small change in systemic hemodynamics, with parallel decreases in arterial pressure and peripheral resistances, and a small, nonsignificant increase in CO. In cirrhotic rats, PCSA induced a decrease of CO to values similar to those of control rats, with an increase in total peripheral resistances. PCSA induced an increase in hepatic arterial blood flow in control and in cirrhotic rats, portal pressure becoming in this latter group not different from that of control rats. Blood flow to splanchnic organs was higher in SO cirrhotic than in SO control animals. Thus portal venous inflow was also increased in SO cirrhotic rats. PCSA induced an increase in portal venous inflow in control rats, which was only significant in cirrhotic rats when expressed as a percentage of CO. In SO control animals, a significant correlation was observed between total peripheral resistances and splanchnic arteriolar resistances and between CO and splanchnic blood flow. These correlations were not observed in cirrhotic rats. These results do not support the hypothesis that hyperdynamic circulation shown by cirrhotic rats is based on increases in splanchnic blood flow and (or) massive portal systemic shunting.     

Digitalis attenuates arterial hypertrophy in experimental hypertension

Several investigators have reported that digitalis administration reduces cardiac hypertrophy in rats with experimental hypertension. To determine whether digitalis similarly affects growth of arteries, we studied young (5- to 14-week-old), male, one-kidney, one-clip hypertensive rats (1K1C; n = 14) and one-kidney normotensive control rats (1K; n = 26). Half of the rats received digoxin (150 mg/kg body wt/day) in chow starting 1-2 weeks before clipping (1K1C-D; 1K-D); the other half were pair-fed (1K1C-C; 1K-C). Serum digoxin levels averaging 488 ng/ml were documented in rats receiving digoxin. After 3-5 weeks of hypertension (conscious tail blood pressures), and at a similar time period in normotensive control rats, we measured direct femoral arterial pressure and weighed standardized segments of the thoracic aorta. At sacrifice body weights of the four groups did not differ. In the one-kidney control rats, mean +/- SE femoral arterial pressure (1K-D, 108 +/- 3; 1K-C, 111 +/- 4, mm Hg), thoracic aortic dry weight (1K-D, 36.6 +/- 0.6; 1K-C, 36.2 +/- 1.1. mg/kg body wt), and aortic water content (1K-D, 62.7 +/- 0.4; 1K-C, 62.4 +/- 0.4, % wet weight) did not differ between rats receiving or not receiving digoxin, respectively. As compared with pooled normotensive control rats, femoral arterial pressure (1K1C-D, 165 +/- 8; 1K1C-C, 153 +/- 5), aortic water content (1K1C-D, 64.8 +/- 0.4; 1K1C-C, 64.9 +/- 0.5), and aortic weight (1K1C-D, 44.8 +/- 2.1; 1K1C-C, 50.1 +/- 1.6) were increased (P less than 0.001) in the one-kidney, one-clip rats, on or off digoxin. Comparison of hypertensive rats receiving to those not receiving digoxin revealed no differences in arterial pressure or aortic water content, but aortic growth was significantly attenuated (-41%, P = 0.02) in the hypertensive rats receiving digoxin. These results provide evidence that digoxin reduces hypertensive arterial growth by a mechanism that does not affect normal growth.     

Direct Fick application for measurement of cardiac output in rat.

The direct Fick procedure for cardiac output determination in rat was validated by simultaneous comparison with electromagnetic flowmeter techniques. Significant coefficients of correlation were obtained between absolute cardiac output values (r = 0.789, P less than 0.001), increases (r = 0.768, P less than 0.001) and decreases (r = 0.672, P less than 0.01) in cardiac output detected by the two methods. As demonstrated in other species, cardiac output values of the Fick procedure in the rat were between 40 and 58% greater than respective electromagnetic flow probe values; however, percent changes in cardiac output obtained by the two methods were similar. The larger values of cardiac output obtained by the direct Fick method may be related, to a great extent, to the distribution of blood flow to the coronary and bronchial circulations. Fick cardiac output measurements were reproducible within rats, and the degree of variation in values among rats was similar to that obtained with the flowmeter procedure. The result indicate that the Fick meth od provides a valid estimation of cardiac output in the rat, with the ability to detect moderate changes (22-36%) in cardiac output.     

Response to nor-epinephrine, isoprenaline and spironolactone derivatives in normal and hypertensive rats

In normal as well as in experimentally hypertensive rats the Blood Pressure and Heart Rate response-time curves are investigated following administration of nor-epinephrine, isoprenaline, spironolactone and K-canrenoate. The results stress the pharmacological effect dependence on integrity of homeostatic blood pressure mechanisms. alpha and beta receptor systems, as well as renine-angiotensine-aldo sterone system show different response-kinetic in mononefrectomized and mononefrectomized plus controlateral renal stenosis rats (Grollmann's model), in comparison with normal. While in normal rats the spironolactones evoke an hypertensive effect in experimentally renal hypertensive rats they reduce pressure to normal values.     

Locomotor Activity and Temperature Rhythms in Cirrhotic and Portal Hypertensive Rats

This study was performed to examine the circadian variations in body temperature and locomotor activity in two rat models of liver damage and portal-systemic collateralization, it is, cirrhosis by common bile duct ligature and portal vein ligation. Locomotor activity and temperature were measured telemetrically, and the degree of portal-systemic shunting was evaluated by the radioactive microsphere technique. In cirrhotic rats a significant increase in portal pressure and portal-systemic shunting occurred, with extensive liver damage and ascitis. These changes were accompanied by a derangement of the activity rhythm (decrease in total activity, night/day ratio and Qp) and an increase in the amplitude of the temperature rhythm. In portal vein-ligated rats, portal vein pressure and portal-systemic shunting increased significantly, with no changes in any of the rhythm parameters analyzed (total, diurnal and nocturnal activity pulses, night/day activity ratio and Qp for activity; mesor, amplitude and free-running period for temperature). The results indicate that liver dysfunction, and not merely portal hypertension or portal-systemic shunting, is the main factor affecting daily rhythms in cirrhotic rats.     

Hemodynamic effects of Salvia miltiorrhiza on cirrhotic rats

Salvia miltiorrhiza (Sm) administration has been shown to reduce hepatic fibrosis in rats. We investigated the hemodynamic effects of Sm on bile duct ligated (BDL) rats. Hemodynamic, histological, and vascular contractile studies were conducted in rats 4 weeks after bile duct ligation. An aqueous extract of Sm (0.2 g twice per day) or vehicle was administered for 4 weeks to BDL rats. Sm treatment in BDL rats significantly reduced histological grades of fibrosis and ameliorated the portal hypertensive state (including portal venous pressure, superior mesenteric artery blood flow, cardiac index, and total peripheral resistance) as compared with vehicle treatment. Moreover, Sm treatment enhanced the vascular sensitivity of mesenteric arteries to phenylephrine in BDL rats. Sm treatment had no effect on plasma biochemical profiles of either BDL or normal rats. Our results suggest that 4-week Sm treatment ameliorates the portal hypertensive state in BDL rats.     

In vivo pressure-flow curve in unilateral rat lung ischemia-reperfusion injury.

The pressure-flow (P-Q) curve has been widely used in many studies to describe the effects of various factors on vascular hemodynamics. It is not clear, however, whether unilateral ischemia-reperfusion (IR) alters the P-Q curve of the rat lung. In this study, we developed an in vivo P-Q curve using the unilateral (left) rat lung before and after IR. Animals were divided into two groups: sham and IR. The protocol of the IR group consisted of three periods: baseline, ischemia, and reperfusion. P-Q curves were obtained by altering blood flow of the left lung during the baseline and the reperfusion periods. The sham group received the same operation without IR procedure. An additional group was used to compare pulmonary blood flow measured by the microsphere and the ultrasonic methods. IR treatment rotated the P-Q curve toward the left, indicating an increase in resistance in the left lung. However, this rotation was not found in the sham group. A significant correlation (r = 0.87, P < 0.01) between percentages of blood flow obtained by the microsphere and ultrasonic methods in both right and left lungs was demonstrated. Therefore, we demonstrated a simple and useful technique to evaluate changes in the P-Q curves caused by IR in the unilateral rat lung model.     

Enhanced formation of PGI2, a potent hypotensive substance, by aortic rings and homogenates of the spontaneously hypertensive rat.

Intact rings and homogenates of aorta from spontaneously hypertensive rats (SHR) contain enhanced capacity over normal rats (NR) to convert arachidonic acid into PGI2. The PGI2 synthetic system in SHR is stimulated to a greater extent than NR by norepinephrine. Indomethacin blocks this stimulation. PGE2 and PGF2alpha were detected in much smaller amounts in homogenates (undetected in rings) but their formation was not enhanced by the hypertensive tissue. The identity of PGI2 was based on 1) direct pharmacological assay on the rat blood pressure. In this system identical vasodepressor responses to PGI2 are observed after intracarotid and intrajugular administration 2) indirectly as 6-keto PGF1alpha isolated after incubation of aortic homogenates with tritiated arachidonic acid and 3) indirectly by GC-MS assay of PGE2, PGF2alpha and 6-keto PGF1alpha formed during incubation of aortic homogenates with excess unlabeled arachidonic acid. These results provide additional support to our recent hypothesis that PGI2, of aortic origin, might actively participate in the regulation of systemic blood pressure. Its enhanced formation by intact hypertensive vascular tissue reflects an increase in the number of enzyme molecules immediately available to the substrate. This could probably be an adaptive response to the elevated levels of catecholamines in the circulation.     

Continuous measurement of aortic blood velocity,after cardiac surgery,by means of an extractable doppler ultrasound probe

A new method has been developed for the continuous measurement of aortic blood velocity in patients following cardiac surgery. Using an extractable Doppler ultrasound probe placed on the ascending aorta, the changes in aortic velocity were recorded up to 24 h postoperatively, in 14 patients undergoing coronary bypass surgery. Volume flow rate is calculated from the mean velocity, the diameter of the aorta and the angle between the ultrasound beam and the direction of the blood flow, by means of an analogue flow calculator. Estimation of aortic flow showed a correlation of r = 0.79 with cardiac output measured by a thermodilution technique. The main advantage of the system is that it allows continuous monitoring of cardiac output, as well as short and long-term trend analyses, during the early postoperative period.     

Impaired generation of prostaglandins from isolated gastric surface epithelial cells in portal hypertensive rats

We studied generation of prostaglandins E2 and 6-keto F1a by surface epithelial cell isolated from the gastric mucosa of portal hypertensive and sham-operated rats. Oxygenated cell suspensions containing 80 +/- 3% of surface epithelial cells were incubated for 30 min at 37 degrees C and the concentration of prostaglandin E2 and 6-keto-prostaglandin F1a in medium was measured by radioimmunoassay. Viability of the cells was assessed with Fast green exclusion at baseline and after 30-min and 60-min incubation. Within 30 minutes the surface epithelial cells obtained from portal hypertensive rats generated 22.0 +/- 1.6 (mean +/- SE) pg prostaglandin E2 and 40.7 +/- 4.7 pg 6-keto prostaglandin F1a, per 10(6) cells. These were significantly less than prostaglandin generation by cells obtained from sham-operated rats. The viability of the surface epithelial cells from portal hypertensive rats was also significantly reduced compared with sham-operated rats after 60 minute incubation. Reduced ability of the surface epithelial cells to generate prostaglandins may be one mechanism for increased susceptibility of portal hypertensive gastric mucosa to injury by noxious agents.     

Endothelin in the splanchnic vascular bed of DOCA-salt hypertensive rats

Vascular capacitance is reduced by endothelin-1 (ET-1) in deoxycorticosterone (DOCA)-salt hypertensive rats. This may contribute to hypertension development. Because the splanchnic blood vessels (especially veins) are important in determining vascular capacitance, we tested the hypothesis that ET-1 levels in the splanchnic vasculature are elevated in hypertensive DOCA-salt compared with normotensive rats. Tissue ET-1 content was measured by ELISA in aorta, vena cava, superior mesenteric artery and vein, and small mesenteric arteries and veins from normotensive sham-operated (sham) and 4-wk DOCA-salt rats. We also determined ET-1 concentration in aortic and portal venous blood (draining the nonhepatic splanchnic organs) in anesthetized and conscious sham and DOCA-salt rats before and after acute blockade of ETB receptor-mediated plasma clearance of ET-1. Results showed a higher ET-1 content in veins than in arteries of similar size. However, ET-1 content was similar in vessels from sham and DOCA-salt rats, except in aorta and superior mesenteric artery, where ET-1 content was greater in DOCA-salt rats. ET-1 concentration was significantly higher in portal venous than in aortic blood, indicating net nonhepatic splanchnic release (nNHSR) of ET-1. However, nNHSR of ET-1 was similar in sham and DOCA-salt rats. Although nNHSR of ET-1 increased significantly after ETB receptor blockade in sham rats, it was completely unchanged in DOCA-salt rats. These data suggest that, despite the absence of ETB receptor-mediated plasma clearance of ET-1, neither the venous peptide content nor the net release of ET-1 is increased in the splanchnic vasculature of DOCA-salt rats. These results argue against the hypothesis that increased venomotor tone in DOCA-salt hypertension is caused by increased ET-1 concentration around splanchnic venous smooth muscle cells.     

Portal hypertension produces an evolutive hepato-intestinal pro- and anti-inflammatory response in the rat

An inflammatory etiopathogeny can be suggested in portal hypertensive enteropathy since infiltration of the intestinal wall by mononuclear cells has been described in this condition. This work was carried out with the intention of shedding light on this matter. Male Wistar rats were divided into 4 control groups and 4 groups with partial portal vein ligation at 1, 2, 3 and 15 months. TNF-alpha, IL-1beta and IL-10 were quantified in liver and ileum by ELISA. CO and NO were measured in splanchnic and systemic vein by spectrophotometry and Griess reaction, respectively. Expression of constitutive and inducible isoforms of NO and HO were assayed by Western blot in liver and ileum. An increased hepatic release of proinflammatory mediators (TNF-alpha, IL-1beta and NO) associated with intestinal release of anti-inflammatory mediators (IL-10, CO) occurs in an early evolutive phase (1 month) of experimental portal hypertension. On the contrary, in the long-term (15 months), the increase in the intestinal release of proinflammatory mediators (TNF-alpha, IL-1beta) is associated with an increase in the hepatic release of anti-inflammatory mediators (IL-10, CO). These results suggest that experimental prehepatic portal hypertension presents changes in the serum and tissular (liver and small bowel) concentrations of mediators which are considered as pro- and anti-inflammatory.     

Splanchnic hemodynamics and gut mucosal-arterial PCO(2) gradient during systemic hypocapnia.

The effects of hypocapnia [arterial PCO(2) (Pa(CO(2))) 15 Torr] on splanchnic hemodynamics and gut mucosal-arterial P(CO(2)) were studied in seven anesthetized ventilated dogs. Ileal mucosal and serosal blood flow were estimated by using laser Doppler flowmetry, mucosal PCO(2) was measured continuously by using capnometric recirculating gas tonometry, and serosal surface PO(2) was assessed by using a polarographic electrode. Hypocapnia was induced by removal of dead space and was maintained for 45 min, followed by 45 min of eucapnia. Mean Pa(CO(2)) at baseline was 38.1 +/- 1.1 (SE) Torr and decreased to 13.8 +/- 1.3 Torr after removal of dead space. Cardiac output and portal blood flow decreased significantly with hypocapnia. Similarly, mucosal and serosal blood flow decreased by 15 +/- 4 and by 34 +/- 7%, respectively. Also, an increase in the mucosal-arterial PCO(2) gradient of 10.7 Torr and a reduction in serosal PO(2) of 30 Torr were observed with hypocapnia (P < 0.01 for both). Hypocapnia caused ileal mucosal and serosal hypoperfusion, with redistribution of flow favoring the mucosa, accompanied by increased PCO(2) gradient and diminished serosal PO(2).     

Echocardiographic measurement of cardiac output in rats

The systematic evaluation of different transthoracic echocardiographic (TTE) methods to determine cardiac output (CO) and the effect of changes in intravascular volume on echocardiographically determined indexes of cardiovascular structure in the rat has not been documented. With the use of 11 Wistar rats, simultaneous echocardiographic and thermodilution measurements of CO were compared at baseline and after blood withdrawal or transfusion at 43 different levels of intravascular volume and using 10 different echocardiographic approaches. The best correlation (r = 0.93; P < 0.0001), least bias (-3 ml/min), and best precision (16 ml/min) between thermodilution and echocardiographic methods were obtained at the level of aortic annulus using pulsed Doppler. In conclusion, CO could be accurately assessed in rats using TTE and pulsed Doppler at the level of the aortic annulus. This annulus was demonstrated to remain stable, but pulmonary annulus, thoracic aorta, mitral valve, and left ventricular diameters were found to be more modifiable during volumic changes.     

Exposure of neonatal rats to carbon monoxide alters cardiac adaptation to aortic constriction.

We tested the hypothesis that a 32-day exposure of newborn rats to 500 ppm carbon monoxide (CO) would alter the adaptive response of the heart to aortic constriction in adulthood. At 110 days of age aortic constriction or sham operations were performed, and hearts were studied 28 days later. Aortic constriction increased left ventricular (LV) mass by 40% over the control value of 611 +/- 27 mg; this adaptive response was not altered by CO exposure. Aortic constriction and CO exposure increased right ventricular (RV) mass by 10 and 11%, respectively, over the control value of 185 +/- 10 mg. The effects of both experimental procedures on RV mass were additive (23%). Peak LV pressure development (dP/dtmax) in vitro increased 29% after aortic constriction in the nonexposed rats. CO exposure blunted the increase in peak LV systolic pressure due to aortic constriction. Maximum positive and negative dP/dtmax decreased by 19% after aortic constriction and were unaffected by CO exposure. The percentage of alpha-myosin heavy chain (MHC) in the ventricles was 94 +/- 2% in the control group and was decreased to 81 +/- 3% by aortic constriction. In contrast, the percentage of alpha-MHC was 87 +/- 2% for CO-exposed rats and was not significantly altered after aortic constriction. In vitro coronary flow was increased 18% in hearts of adult rats exposed to CO as neonates. Exposure of neonatal rats to CO induced chronic adaptations in the myocardium, some of which became evident in adulthood only when hearts were challenged by aortic constriction.     

Autoregulation of renal medullary blood flow in rabbits

We examined the extent of renal medullary blood flow (MBF) autoregulation in pentobarbital-anesthetized rabbits. Two methods for altering renal arterial pressure (RAP) were compared: the conventional method of graded suprarenal aortic occlusion and an extracorporeal circuit that allows RAP to be increased above systemic arterial pressure. Changes in MBF were estimated by laser-Doppler flowmetry, which appears to predominantly reflect erythrocyte velocity, rather than flow, in the kidney. We compared responses using a dual-fiber needle probe held in place by a micromanipulator, with responses from a single-fiber probe anchored to the renal capsule, to test whether RAP-induced changes in kidney volume confound medullary laser-Doppler flux (MLDF) measurements. MLDF responses were similar for both probe types and both methods for altering RAP. MLDF changed little as RAP was altered from 50 to >or=170 mmHg (24 +/- 22% change). Within the same RAP range, RBF increased by 296 +/- 48%. Urine flow and sodium excretion also increased with increasing RAP. Thus pressure diuresis/natriuresis proceeds in the absence of measurable increases in medullary erythrocyte velocity estimated by laser-Doppler flowmetry. These data do not, however, exclude the possibility that MBF is increased with increasing RAP in this model, because vasa recta recruitment may occur.     

肾上腺髓质素的降压作用及其在组织中分布的观察

本实验用电生理和免疫组化方法观察了肾上腺髓质素（ａｄｒｅｎｏｍｅｄｕｌｌｉｎ,ＡＭ）对正常和高血压大鼠血压的影响和对主动脉环张力的作用以及ＡＭ在一些组织中的分布特点。结果显示ＡＭ对高血压大鼠的降压幅度远大于正常鼠;对主动脉环张力的降低作用具有剂量依赖性和内皮依赖性;ＡＭ免疫组化反应除在嗜铬细胞瘤和肾上腺髓质细胞内呈阳性反应外,在肺、肾和心肌组织内都显示出不同程度的阳性。提示ＡＭ在机体血压和循环调节中具有重要作用。     

Effects of chronic portal hypertension on small heat-shock proteins in mesenteric arteries

Previous studies have shown that impaired vasoconstrictor function in chronic portal hypertension is mediated via cAMP-dependent events. Recent data have implicated two small heat-shock proteins (HSP), namely HSP20 and HSP27, in the regulation of vascular tone. Phosphorylation of HSP20 is associated with vasorelaxation, whereas phosphorylation of HSP27 is associated with vasoconstriction. We hypothesized that alterations in the expression and/or phosphorylation of small HSPs may play a role in impaired vasoconstriction in portal hypertension. A rat model of prehepatic chronic portal hypertension was used. Studies were conducted in small mesenteric arteries isolated from normal and portal hypertensive rats. Protein levels of HSP20 and HSP27 were detected by Western blot analysis. Protein phosphorylation was analyzed by isoelectric focusing. HSP20 mRNA expression was determined by RT-PCR. To examine the role of cAMP in the regulation of small HSP phosphorylation and expression, we treated both normal and portal hypertensive vessels with a PKA inhibitor Rp-cAMPS. We found both an increased HSP20 phosphorylation and a decreased HPS20 protein level in portal hypertension, both of which were restored to normal by PKA inhibition. However, PKA did not change HSP20 mRNA expression. We conclude that decreased HSP20 protein level is mediated by cAMP-dependent pathway and that impaired vasoconstrictor function in portal hypertension may be partially explained by decreased expression of HSP20. We also suggest that the phosphorylation of HSP20 by PKA may alter HSP20 turnover.