BEPI LAB 1: BAROREFLEX CONTROL OF CARDIOVASCULAR FUNCTION

THE EXPERIMENTAL PLATFORM

These experiments will be conducted on a large-scale simulation of the cardiovascular, respiratory, and
renal systems called SimBioSys. The simulation simultaneously solves hundreds of differential equations
that simulate physiological systems. We will concentrate on the cardiovascular portions of the simulation in
this lab. Provisions exist for delivering many different drugs, infusing saline or drawing blood, and both
measuring and manipulating several intrinsic cardiovascular parameters.

OVERVIEW/STARTING THE SIMULATION

1. Click on the link in Blackboard to save a ZIP file with program to your computer. Open the ZIP file, and
click on the “sbs3labs” icon; this will actually save the simulation to your hard drive. Once this step is
finished, click on the “sbs3labs” icon that looks like a person in a white shirt with a heart where the head
should be. This will open the simulation.

2. Click “OK” to bypass the sign-in form.

3. Click on the “File” menu, then “Load Physiology State.”

4. Double click on the “sbs3template2013” file, and answer “yes” to the dialog box that appears. A panel
will appear across the top of the screen that has aortic pressure (top) and heart rate (bottom) plotted
against time.

5. Below the aortic pressure and heart rate panel of tracings, you should see some “Viewer” boxes. Box 1
will be very important for everything you do in this lab, so expand that box until you can see all variables
and output values. (If a “Viewer 2” box also opened when you started the stimulation, you can click the
red “X” in the corner to it—you won’t need it today.)

6. The “Drug and Fluid Infusor” and “Blood Withdrawal” viewer boxes should also be open, down next to the
Viewer 1 box. If they are not, go over to the “Tools” menu box in the lower left corner of the screen and
check the box next to the “Blood Withdrawal” option, and also check the box next to the “Drug and Fluid
Infusor” option. This should open those 2 additional boxes for you, making their options now available;
you will need these 2 boxes later on in the lab. (If these boxes disappear when you periodically need to
re-load the physiology state to start a new simulation, that’s ok—you now know how to make them re-
appear.)

7. To better see all data tracings and boxes, collapse the left-side panel that contains the “Tools” and
“Curriculum,” and expand the tracing panels and boxes until you can see everything comfortably.

8. During the lab, you may want to open a Word or PowerPoint document, because there will be several
moments where you will want/need to do a screen capture and then paste that image into the open
document.

9. There are some handy features in the simulation:

• First, you can click on the camera icon on the data panel or in the Viewer 1 box to capture (in gray)
the values at a specific moment while letting the simulation continue to run.
 • Second, you can click on the “Simulation” button at the top of the screen, then go to “Set Simulator
Speed,” and then click on “Pause.” This gives you a frozen screen, in case you want to take a
screen capture image or manually record values at that moment before continuing on with a
particular lab protocol. When you want to resume simulation, you can go back to this button and
click the speed at which you would like to have the simulation run. (We recommend 4x.)

EXERCISE 1: CAROTID SINUS ISOLATION

We begin the lab by studying the neural regulation of the circulation through the arterial baroreflex. (Once
we are familiar with this classic cardiovascular control system, we will then proceed to manipulate various
internal elements of the system, as well as external stimuli, to further explore how the baroreflex
compensates for changes in pressure and what happens in cases of baroreflex dysfunction or failure.)

Usually, carotid sinus pressure (i.e., the baroreflex stimulus) and mean arterial pressure (i.e., the baroreflex
response) are similar; indeed, the reflex depends on the assumption that carotid sinus pressure is
representative of arterial pressure in general. However, our simulation permits us to uncouple mean arterial
pressure from carotid sinus pressure (isolating the carotid sinus from the rest of the circulation), enabling us
to manually clamp carotid sinus baroreceptor pressure at any level we wish and then monitor how the rest of
the system responds. The brain in the simulation thinks that this carotid sinus input is the actual arterial
pressure and compensates accordingly, using the autonomic nervous system. (We should keep in mind that
the aortic baroreceptors are still intact in this simulation, and because they are still sensing actual arterial
pressure, they will blunt the effects of our changes in carotid sinus pressure somewhat.)

If you haven’t not already done so, click “File” > “Load Physiology State” and get your viewer windows all set
(see p.1 for instructions on how to do this). Wait ~ 20 seconds for the system to come to an initial steady
state. For this protocol, you will set carotid sinus pressure at a high level (160 mmHg) and, after recording
the relevant data, you will then incrementally reduce carotid sinus pressure to progressively lower levels
(recording data below as you go). To set carotid sinus pressure: click on the lock icon for the sinus
pressure entry. This will “unlock” carotid sinus pressure, meaning that it can no longer be controlled/affected
by the simulation—you will now control it instead.1 Click on the numeric value of carotid sinus pressure. It
will then become available for setting. Type “160.” (do not forget the decimal point at the end!) and then hit
“Enter.” Allow the simulation to come to a new steady state (may take 30 seconds or so), and then record
the relevant variable values in the “160 mmHg” row of the chart below.
• Since the values will vary somewhat from second to second, you may find it useful to pause the simulation
before recording these values. This is done by clicking on the “Simulation” menu > “Set Simulator Speed” > “Pause.”
When you want to restart the simulation, return to this menu option and click on the speed you want (4x is
recommended). After you click on 4X, “yes” your way past the warning message and the simulation will begin where
it left off.

Carotid Systemic Central

Parasympathetic Sympathetic Mean aortic Cardiac
sinus HR vascular venous
tone tone pressure output
pressure resistance pressure
160 mmHg I - ooo oooo
27 26 1.10 24.6 -
2-3
140 mmHg 26
0-730 • - ooo
31 I -22 22-3 -2-2
120 mmHg
0-333 0 - ooo
52 41 1.82 23-6 -2-3
110 mmHg
0-245 0-0%668 57 2-42 24-4 -2-1

1
“Locked” and “Unlocked” variables. The most common misunderstanding in running SBS3 is the concept of “locking” a variable.
When a variable is “locked,” it is locked into the simulation and controlled by the simulation. It is not fixed! If you want to fix or set a
variable yourself, unlock it! It will stay at whatever value it had when you unlocked it until you click on that value and either substitute your
own value (followed by “enter”) or click on the variable’s right arrow to set its value with a slider.
 100 mmHg 0.3225 a-
177 78 914 3.31 28-6 -
l - o
95 mmHg 0-368 0-318 92 126 3-86 32-7 -0-7
90 mmHg
80 mmHg
0-353 0-467 V4 161 4.31 37.5 -0-7
0-222 0-642 155 166 3-87 43 y -
- O -
G

Without resetting the simulation in any way, repeat this process for each level of sinus pressure in the
table (in descending order). At each carotid sinus pressure, permit the simulation to reach a steady state
(~ 30 seconds) before recording data in the chart. Don’t worry about the slow (and small) changes in some
variables once arterial pressure has leveled off. (Please note that you aren’t collecting data for SV, LVEDV,
or LVESV; these values tend to respond less naturally in this simulation, so you won’t be recording them.)

If you need to re-run the experiment, do not use the “Reset Physiological State” under the “Simulation”
menu. Instead, go to “File” > “Load Physiology State.” Please do not “Save Physiological State” under
any circumstances!
EXERCISE 2. CIRCULATORY EFFECTS OF DRUG INFUSIONS

CIRCULATORY EFFECTS OF NOREPINEPHRINE ADMINISTRATION

Now, you will increase arterial pressure with a dangerously large infusion of the sympathetic adrenergic
neurotransmitter norepinephrine (NE), simulating a gross clinical error. NE will activate b1 adrenergic
receptors on cardiac pacemaker cells in the SA node (leading to an increase in heart rate) and on cardiac
ventricular muscle (leading to an increase in cardiac contractility). NE will also bind to a1 adrenergic
receptors on vascular smooth muscle in both the arterial and venous sides of the circulation (increasing
systemic vascular resistance and decreasing venous capacitance, respectively).

Begin by going to “File” > “Load Physiology State.” While the simulation is taking about 20 seconds to find
an initial steady state, make sure that you have your “Drug and Fluid Infusor” viewer box visible for this next
part. (See p.1 for instructions on how to do that.) To infuse NE, click on the “+” in the Drug Infusor box and
scroll down to “norepinephrine” in the pharmacy and click on it. You are only permitted to deliver NE as an
infusion, so you won’t make a choice of mode of delivery on the left side of this window. So just click “Add
Drug.” Note: this does not infuse NE in the simulation yet—it just adds it to the bank of drugs you can give.

You will now set up the infusion rate for the norepinephrine. Being careful not to hit the red button (which
would cause the norepinephrine infusion to start), click the arrow in the norepinephrine box. On the dropdown
menu, click on dose, and increase it to 6.00 µg/kg/min. Close the dropdown menu.

Before you start the infusion, pause the simulation (or hit the camera icon in the Viewer 1 box) and record
baseline data in the chart below. At this point, you will now click on the red button on the “norepinephrine” line
of the Drug Infusor box. This will begin the infusion. Pause the stimulation after it has run for ~50 seconds in
“simulation time” (from one x-axis hashmark on the time scale to the next). Record the values of the variables
in the next column on the chart. (After ~50 seconds, the high dose of this powerful adrenergic agonist causes
severe instabilities in the model, just as it would in a person, so you wouldn’t use them.)

Pre-infusion Response after 50 sec

baseline NE infusion

Parasympathetic tone
0.343 0-591
 Sympathetic tone
0.190 0.005
HR
78 55
Mean aortic pressure
101 122
Cardiac output
3. 55 2.85
Systemic vascular resist.
28.4 44 / .

Central venous pressure

- 0.6 - 0.
/

You may notice that parasympathetic tone does not seem to change very much or very rapidly. This violates
the description of parasympathetic effects that you have heard in all of your lectures. Unfortunately, neither
models nor humans are perfect, and this is a property of the simulation model that needs some work.
Increasing the speed of the parasympathetic response to the heart causes this model to become unstable.
Therefore, it is deliberately set to be less responsive than it would be in a real person.

Re-start a new simulation by clicking “File” > “Load Physiology State” and allow the simulation to come to a
new initial baseline. (You may need to reinstate the drug and dose because you loaded a new simulation.)
You are going to run the same NE protocol again, except that this time you are going to prevent the
autonomic nervous system from compensating for the body’s response to the NE overdose—you’ll get to
see what happens when the efferent arm of the baroreflex is disabled. We do this by “unlocking” the
sympathetic and parasympathetic tone values. Click on the locks on the left sides of their respective rows in
the viewer box. When unlocked, the autonomic tone will stay clamped at normal resting levels, unable to
respond to any stimulus. Pause the simulation and record all baseline values (including the unlocked
parasympathetic and sympathetic values) in the chart below. If you have not yet re-loaded the Drug Infusor
with NE at 6.0 µg/kg/min, do that now. Un-pause the simulation and begin the NE infusion, again pausing
after ~ 50 sec. Record your data in the chart.
uncompensated Bano receptors
Response after 50 sec
NE infusion
Pre-infusion baseline

Unlocked at baseline value: Unlocked at baseline value:

Parasympathetic tone
0-345 0-345
Unlocked at baseline value: Unlocked at baseline value:
Sympathetic tone
0-189 o -
289
HR
77 88
Mean aortic pressure
too 199
Cardiac output
3- 54 4.07
Systemic vascular resist.
28.4 49 I .

Central venous pressure

- 0-6 - O -
3

CIRCULATORY EFFECTS OF NITROGLYCERIN ADMINISTRATION

Ent vasodilator w/ no
cardiac
Click “File” > “Load Physiology State.” You will now set up a similar kind of drug infusion scenario, only this effect
time you will be administering an overdose of nitroglycerin, a potent vasodilator with no cardiac effects. In

o
the “Drug and Fluid Infusor” box, scroll down to “nitroglycerin” and click “Add Drug.” Increase the dose up to
200 µg/min (continuous infusion), but don’t start the infusion yet. Pause the simulation after it has reached a
stable, pre-infusion baseline and record those values in the chart below. Now un-pause the simulation and
click on the red button next to “nitroglycerin,” allowing the infusion to proceed for 200 seconds of “simulation
time” (4 hashmarks along the x-axis time scale). Follow the effects of the drug on the arterial pressure &
heart rate (tracings) and on the other variables in the data table. Pause the simulation after 200 seconds
and record all data values in the middle column on the chart.

Response after 200 sec

nitroglycerin infusion
Pre-infusion baseline

Parasympathetic tone o -
343 0-323

5
 Sympathetic tone 192
0.444
o -

HR
78 115
Mean aortic pressure
101 91
Cardiac output
3- 57 4.70
Systemic vascular resist.
28.5 19.4
Central venous pressure
-
G. 6 - o -
3

Click “File” > “Load Physiology State” and allow the simulation to come to a new baseline, and (if necessary)

0
reload nitroglycerin at 200 µg/kg/min back into the “Drug Infusor” window. You are going to run the same
nitroglycerin protocol again, except that this time you are going to unlock the parasympathetic and
sympathetic responses, thereby preventing the baroreflex from compensating for an overdose of a potent
vasodilator, much like you did in the NE infusion protocol. After you unlock parasympathetic and
sympathetic tones, pause the simulation and record all baseline values (including the unlocked
parasympathetic and sympathetic values) in the chart below. Then un-pause the simulation and start the
nitroglycerin infusion, allowing it to proceed for 200 seconds, as you did before. After 200 seconds, pause
the simulation again to record data values in the second column of the chart below.

Response after 200 sec

nitroglycerin infusion
Pre-infusion baseline

Unlocked at baseline value: Unlocked at baseline value:

Parasympathetic tone
0.345 0.345
Unlocked at baseline value: Unlocked at baseline value:
Sympathetic tone
0.186 o- 186
HR
77 77
Mean aortic pressure
too 36
Cardiac output
3. 54 2- 80
Systemic vascular resist.
28.3 15-5
Central venous pressure
- o. 6 -2-3

6
 EXERCISE 3. EXAMINING PARASYMPATHETIC AND SYMPATHETIC EFFECTS

Now, you will study the effects of sympathetic and parasympathetic control of the circulation separately. Keep
in mind that changes in parasympathetic activity in this simulation are a little more sluggish than you’d see in
a real human. However, the cardiac and systemic effects of sympathetic tone are nicely distinguished, so the
sympathetic tone portion of this exercise is a little more extensive.

PARASYMPATHETIC TONE

In this exercise, we are going to study the effect of vagal nerve stimulation on heart rate. (The vagus nerve
carries ~75% of all parasympathetic outflow to the body.) Start the simulation (“File” > “Load Physiology
State”) and allow it settle down into a nice steady state baseline. To prevent the baroreceptor system from
“muddying the waters” by compensating for the increasing parasympathetic tone, you will first start out by
unlocking sympathetic tone to clamp it as baseline levels. Then you will simulate increasing levels of vagal
stimulation by unlocking parasympathetic tone and successively setting parasympathetic tone at levels of
0.0, 0.4, and 0.8. After you manually set parasympathetic tone at each of these levels, wait for a new steady
state, and then record heart rate on the chart below before you move to the next parasympathetic tone level.

Parasympathetic Tone: 0.0 0.4 0.8 0.8 + atropine

Heart Rate: 126

To 39 116
Remember that vagal stimulation works when the nerve ending releases acetylcholine, which then binds to
muscarinic receptors on the tissue. Continue “stimulating the vagus” at the 0.8 level, and now add the
PEE
muscarinic receptor antagonist (blocker) atropine to the Drug Infusor. You only want to deliver this drug once,
a-
but the simulation will not let you give less than 2 doses. However, you will notice that the interval between
the doses is so large that, in effect, only one dose will be given during the period we are studying. Set the
dose of atropine to 2 doses of 2.0 mg each, at a 30-minute internal, and then administer the dose. Record the
new heart rate in the last column of the chart.

SYMPATHETIC TONE

Re-start the simulation (“File” > “Load Physiology State”) and wait for a good steady state baseline. Next, you
will unlock parasympathetic tone (to prevent it from compensating for any upcoming changes) and then
unlock sympathetic tone (so that you can manually control it). Once you have unlocked both of those
variables, record baseline cardiovascular values in the 1st column of the chart on the next page. Now set
sympathetic tone to 0.05. Wait for a new steady state, and record data in the 2nd column of the chart. Now
manually set SNS tone to 0.5, wait for steady state, and record data values in the 3rd column of the chart.
Maintaining this high level of sympathetic stimulation, and without resetting the simulation, you are
going to block the beta (b)-adrenergic receptors in the heart, and then you will add a blockade of the alpha
(a)-adrenergic receptors in the peripheral circulation. To do this, you will need to first administer the
betablocker esmolol as an infusion at 300 µg/kg/min. (Esmolol blocks beta-adrenergic receptors.) Wait for
the system to come to a new steady state after you give the infusion, and record that data in the 4th column of

e-
the chart. While still continuing the esmolol infusion, add the alpha-blocker phentolamine to the Drug
Infusor at a dose of 25 mg (2 doses but 30 minutes apart—this is effective one dose on your timescale). Give
the dose of phentolamine. (Phentolamine blocks only a-adrenergic receptors, which are found in vascular
smooth muscle but not the heart.) Once the system has come to a steady state, record the data in the last
column of the chart.

7
end with lol is beta blocker
Anything a
 SNS = 0.5 + SNS = 0.5 +
b block only b block + a block
Baseline SNS tone = 0.05 SNS tone = 0.5

Unlocked at baseline Unlocked at baseline Unlocked at baseline Unlocked at baseline Unlocked at baseline
Parasympathetic tone
value: value: value: value: value:
0-343 0-343 8-343 0-343 0-345
Unlocked at baseline
Sympathetic tone 0.05 0.5 0.5 0.5
value:
0.190
HR
TO 58 121 Go 56
Mean aortic pressure
lol 59 172 123 65
Cardiac output
3.54 2-46 4.64 3-22 2-60
Systemic vascular resist.
28.6 24.8 37.1 38.1 25.4
Central venous pressure
- 0.6 -1.5 -0-3 0.3 -2-0

EXERCISE 4: HEMORRHAGE

Re-start the simulation (“File” > “Load Physiology State”). In this exercise, you are going to study the effect of
withdrawing blood from the circulation, first with—and then without—compensation by the baroreflex. From
the “Tools” window on the left side of your screen, click on “Blood Withdrawal.” Set the rate at its maximum.
Set the volume at 750 ml. After recording pre-hemorrhage baseline data values in first column of the chart
below (and with sympathetic and parasympathetic variables still locked) press the “Withdraw Blood” button on
the withdraw tool. A minute or so after the full 750 ml has been withdrawn, pause the simulation and record
your results in the second column of the chart. Now re-load the simulation and repeat this experiment,
this time withdrawing 1500 ml of blood. Record your results in the third column of the chart. (For some
perspective, note that blood donation at a blood drive only withdraws 500 ml from the body!) Finally, repeat
this experiment two more times, once at each level of hemorrhage (750 ml and 1500 ml), this time with the
sympathetic and parasympathetic tones unlocked at baseline levels, remembering to re-start the
simulation each time. Record your data in the last 2 columns of the chart.

Pre-hemorrhage
baseline
750 ml loss 1500 ml loss 750 ml loss 1500 ml loss

Unlocked at: Unlocked at:

0-120 02000
Parasympathetic tone 0.344 0-3450-345
Unlocked at: Unlocked at:
Sympathetic tone
191 1.000
G. 0-542 0.191 0-192
 HR
77 157 Zoo
-

77 78
Mean aortic pressure
lol 92 34 37 17
Cardiac output 3. 57 2.32 0-78 1.43 0.85
Systemic vascular resist.
28.4 37 -7 49.5 283 26.9
Central venous pressure -
0.5 -2.6 -4^8 -3-2 -6.5

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7

EXERCISE 5: GANGLIONIC BLOCKADE

Re-start the simulation (“File” > “Load Physiology State”). In this final exercise, you are going to study the
effect of inhibiting autonomic outflow to the cardiovascular system via blockade of all autonomic ganglia. After
recording your baseline data values in the chart below, add trimethaphan (a short-duration ganglionic blocker)
-
to the Drug Infusor window. Keeping the dose and infusion rate at default values, administer the trimethaphan
and allow the simulation to run for ~200 seconds (according to the x-axis time scale). Pause the simulation
and record your data in the chart.

Response after 200 sec

trimethaphan infusion
Pre-infusion baseline

0-344
Parasympathetic tone
o ooo
-

Sympathetic tone
0.192 oooh
HR
77 loss
Mean aortic pressure
101 75
3- 56 3.22
Cardiac output

Systemic vascular resist.

28.3 24
Central venous pressure -
0.5 -1-8
8