Limits...
The combined effects of sidestream smoke extracts and glycated serum albumin on endothelial cells and platelets.

Rubenstein DA, Morton BE, Yin W - Cardiovasc Diabetol (2010)

Bottom Line: In general, the endothelial cell culture conditions were reduced in the presence of AGE and SHS.Nicotine, did not play a role in this reduction.This study also provides a new experimental technique to monitor platelet-endothelial cell interactions.

View Article: PubMed Central - HTML - PubMed

Affiliation: School of Mechanical and Aerospace Engineering, Oklahoma State University, Stillwater, USA. david.rubenstein@okstate.edu

ABSTRACT

Background: The purpose of this study was to test the hypothesis that sidestream tobacco smoke extracts would inhibit the culture of endothelial cells and enhance platelet aggregation under diabetic vascular conditions. Sidestream tobacco smoke and advanced glycation end products are known cardiovascular risk factors and we aimed to determine the combined interaction between these two risk factors to promote cardiovascular diseases associated with diabetes.

Methods: Human umbilical vein endothelial cells were cultured in the presence of sidestream tobacco smoke extracts (SHS) or nicotine and glycated albumin (AGE) or non-glycated albumin. After 3 days, endothelial cell viability and density were investigated. Platelets were also incubated with these compounds for up to 6 hours. Platelet aggregation and the surface expression of CD41 and CD62P were examined. In some experiments, platelets were added to the endothelial cell culture to determine if an interaction between platelets and endothelial cells occurs that can alter the responses to SHS or AGE.

Results: In general, the endothelial cell culture conditions were reduced in the presence of AGE and SHS. Nicotine, did not play a role in this reduction. Platelet aggregation proceeded faster in the presence of AGE and SHS. Interestingly, with the combined culture of endothelial cells and platelets, the endothelial cell culture conditions were improved and the platelet functional changes were diminished in the presence of SHS and AGE, as compared with the individual incubations.

Conclusions: Our data suggests that diabetics that are exposed to SHS may have a higher likelihood for cardiovascular disease development through a diminished endothelial cell viability and an increased platelet activity, which are partially mediated by CD41 and not CD62P. This study provides support for an increased cardiovascular risk for diabetic patients that are exposed to SHS. This study also provides a new experimental technique to monitor platelet-endothelial cell interactions.

Show MeSH

Related in: MedlinePlus

Surface expression of GPIIb (CD41, A) or P-selectin (CD62P, B) on the platelet membrane after the incubation of SHS or nicotine with AGE or BSA. Data are the mean + SEM of 4 independent experiments. * differs from incubation with BSA (t-test, P < 0.05).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC2909174&req=5

Figure 3: Surface expression of GPIIb (CD41, A) or P-selectin (CD62P, B) on the platelet membrane after the incubation of SHS or nicotine with AGE or BSA. Data are the mean + SEM of 4 independent experiments. * differs from incubation with BSA (t-test, P < 0.05).

Mentions: The responses of platelets to individual incubations of SHS, nicotine or glycated albumin have previously been reported by us and were therefore not investigated here[10,11,15]. Instead we investigated the combined effects of SHS extract and glycated albumin incubation on platelet aggregation and the surface expression of CD41. Generally, after exposure to SHS extracts the rate of platelet aggregation was enhanced as compared to control samples and the paired non-glycated albumin controls (Figure 2A). With the addition of pure nicotine instead of SHS extracts, there was a general attenuation in the aggregation rate approaching that of control conditions. Interestingly, the percent change of aggregation was generally not affected by the incubation of SHS extracts, nicotine, glycated albumin or non-glycated albumin (Figure 2B). Combined, the aggregation data suggests that platelets that are exposed to SHS extracts aggregate faster, but the quantity of platelets that participate in aggregation remains the same, independent of the culture additive (SHS, AGE or nicotine). Also, nicotine returns that aggregation rate back to control levels, independent of albumin glycation. Flow cytometry was used to confirm the aggregation findings and investigate a possible mechanism for the changes in aggregation rate and percent change of aggregation. In general, the trend in GPIIb expression mimics the data found for the platelet aggregation rate (Figure 3A). With the addition of SHS to the culture media there is an enhanced surface expression of CD41, which was attenuated with the addition of nicotine. We also investigated the expression of CD62P and saw minimal changes to the expression of P-selectin, even in the presence of SHS or nicotine (Figure 3B).


The combined effects of sidestream smoke extracts and glycated serum albumin on endothelial cells and platelets.

Rubenstein DA, Morton BE, Yin W - Cardiovasc Diabetol (2010)

Surface expression of GPIIb (CD41, A) or P-selectin (CD62P, B) on the platelet membrane after the incubation of SHS or nicotine with AGE or BSA. Data are the mean + SEM of 4 independent experiments. * differs from incubation with BSA (t-test, P < 0.05).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2909174&req=5

Figure 3: Surface expression of GPIIb (CD41, A) or P-selectin (CD62P, B) on the platelet membrane after the incubation of SHS or nicotine with AGE or BSA. Data are the mean + SEM of 4 independent experiments. * differs from incubation with BSA (t-test, P < 0.05).
Mentions: The responses of platelets to individual incubations of SHS, nicotine or glycated albumin have previously been reported by us and were therefore not investigated here[10,11,15]. Instead we investigated the combined effects of SHS extract and glycated albumin incubation on platelet aggregation and the surface expression of CD41. Generally, after exposure to SHS extracts the rate of platelet aggregation was enhanced as compared to control samples and the paired non-glycated albumin controls (Figure 2A). With the addition of pure nicotine instead of SHS extracts, there was a general attenuation in the aggregation rate approaching that of control conditions. Interestingly, the percent change of aggregation was generally not affected by the incubation of SHS extracts, nicotine, glycated albumin or non-glycated albumin (Figure 2B). Combined, the aggregation data suggests that platelets that are exposed to SHS extracts aggregate faster, but the quantity of platelets that participate in aggregation remains the same, independent of the culture additive (SHS, AGE or nicotine). Also, nicotine returns that aggregation rate back to control levels, independent of albumin glycation. Flow cytometry was used to confirm the aggregation findings and investigate a possible mechanism for the changes in aggregation rate and percent change of aggregation. In general, the trend in GPIIb expression mimics the data found for the platelet aggregation rate (Figure 3A). With the addition of SHS to the culture media there is an enhanced surface expression of CD41, which was attenuated with the addition of nicotine. We also investigated the expression of CD62P and saw minimal changes to the expression of P-selectin, even in the presence of SHS or nicotine (Figure 3B).

Bottom Line: In general, the endothelial cell culture conditions were reduced in the presence of AGE and SHS.Nicotine, did not play a role in this reduction.This study also provides a new experimental technique to monitor platelet-endothelial cell interactions.

View Article: PubMed Central - HTML - PubMed

Affiliation: School of Mechanical and Aerospace Engineering, Oklahoma State University, Stillwater, USA. david.rubenstein@okstate.edu

ABSTRACT

Background: The purpose of this study was to test the hypothesis that sidestream tobacco smoke extracts would inhibit the culture of endothelial cells and enhance platelet aggregation under diabetic vascular conditions. Sidestream tobacco smoke and advanced glycation end products are known cardiovascular risk factors and we aimed to determine the combined interaction between these two risk factors to promote cardiovascular diseases associated with diabetes.

Methods: Human umbilical vein endothelial cells were cultured in the presence of sidestream tobacco smoke extracts (SHS) or nicotine and glycated albumin (AGE) or non-glycated albumin. After 3 days, endothelial cell viability and density were investigated. Platelets were also incubated with these compounds for up to 6 hours. Platelet aggregation and the surface expression of CD41 and CD62P were examined. In some experiments, platelets were added to the endothelial cell culture to determine if an interaction between platelets and endothelial cells occurs that can alter the responses to SHS or AGE.

Results: In general, the endothelial cell culture conditions were reduced in the presence of AGE and SHS. Nicotine, did not play a role in this reduction. Platelet aggregation proceeded faster in the presence of AGE and SHS. Interestingly, with the combined culture of endothelial cells and platelets, the endothelial cell culture conditions were improved and the platelet functional changes were diminished in the presence of SHS and AGE, as compared with the individual incubations.

Conclusions: Our data suggests that diabetics that are exposed to SHS may have a higher likelihood for cardiovascular disease development through a diminished endothelial cell viability and an increased platelet activity, which are partially mediated by CD41 and not CD62P. This study provides support for an increased cardiovascular risk for diabetic patients that are exposed to SHS. This study also provides a new experimental technique to monitor platelet-endothelial cell interactions.

Show MeSH
Related in: MedlinePlus