Pingnian He, M.D., Ph.D.
	Professor of Physiology and Pharmacology Medical Degree: Tianjin Medical University, People’s Republic of China. Graduate Training: University of California, Davis. Fellowship: University of California, Davis.
	Office: 3072B-HSN Lab: 3075-HSN PO Box 9229 Morgantown, WV 26506	Email: phe@hsc.wvu.edu Phone: 304-293-1515 Fax: 304-293-3850

Research Interests:

The long-term goal of our research is to investigate the cellular mechanisms that regulate permeability in intact microvessels. Accumulated clinical and experimental evidence indicate that increased vascular permeability during inflammation is the initiating event for a variety of cardiovascular diseases, such as atherosclerosis, diabetes, as well as promoting tumor growth and tumor metastasis. A better understanding of the mechanisms that regulate microvessel permeability is crucial to defining the pathogenesis of many disease conditions, and aiding in the development of novel therapeutic approaches.

Description of Research:

• Signaling mechanisms of microvessel permeability regulation during inflammation. This includes the investigations of the roles of endothelial [Ca²⁺]_i, nitric oxide, caveolin-1, as well as cAMP and cGMP-dependent signaling pathways in the regulation of microvessel permeability.
• Leukocyte/platelet/endothelial cell interaction, the anticipated adhesion molecules and signaling cascade, and their roles in the regulation of microvessel permeability.
• The roles of cytoskeleton and adherent junctions in the regulation of microvessel permeability in intact microvessels.
• The roles of shear stress and the glycocalyx layer in the regulation of vascular functions.
• Lyme disease and microcirculation: Spirochete Borrelia burgdorferi (Bb) interaction with endothelium and Bb-induced vascular inflammatory reaction in intact microvessels.

Approaches:

Individually perfused intact microvessel is the main technique employed in our laboratory. This technique provides us a powerful tool to conduct quantitative assessments of the properties of the vascular walls for fluid and solute transports and to explore the cellular and molecular mechanisms in intact microvessels. Our combined fluorescence microscopy, confocal microscopy, and electron microscopy studies with quantitative measurements of permeability coefficients in intact microvessels bridges the studies of using whole animal, organ, or vascular beds and studies of using cultured endothelial cells in vitro.

Techniques:

• Quantitative measurements of permeability coefficients: Measurement of hydraulic conductivity, Lp (video) and permeability coefficients.
• Measurements of endothelial [Ca²⁺]_i, before and after the vessel was exposed to inflammatory mediators using fluorescence imaging (movie) and photometry system.
• Measurements of endothelial nitric oxide production (Images).
• Three-dimensional visualization of PAF-induced vascular leakage sites in an intact microvessel (animation of confocal images ).
• Three-dimensional visualization of VE-Cadherin distribution in a venular microvessel.
• Confocal imaging of vascular structure: illustrating endothelial structures in a venular wall using combined reflectance (silver staining to outline endothelial clefts) and fluorescence (nuclei staining) confocal images (animated image 1, animated image 2).
• Gene delivery and protein expression in intact microvessels.

Figure 1. Measurement of hydraulic conductivity in an individually perfused intact microvessel. A rat mesenteric venule (diameter 40 µm) was cannulated with a glass micropipette and perfused with a Ringer-albumin solution containing red blood cells as markers. Certain pressure was applied through the pipette and to the microvessel wall. During the measurement, the vessel was occluded downstream with a glass rod, and the velocity of the marker cells were measured. The hydraulic conductivity was calculated based on the velocity of the marker cell movement, the vessel diameter, vessel length, and the perfusion pressure (see equation). View video from Media Library

Figure 2. Confocal reflectance and fluorescence imaging of a post-capillary venule. The endothelial cell clefts (green color) were delineated with silver nitrate staining and the images were acquired with reflectance. The endothelial nuclei (red) were stained with fluorescent nucleic acid dye YO-PRO-1. This image is a projection of the half cylinder of the vessel. Click the image to see the three-dimensional structures of endothelial cells and endothelial nuclei in this intact veunular microvessel. Ref: He, P. and R. H. Adamson. Visualization of endothelial clefts and nuclei in living microvessels with combined reflectance and fluorescence confocal microscopy. Microcirculation 2: 267-276, 1995.

Research Support:

• NIH/NHLBI  RO1 (Pingnian He, Principal Investigator)
  Title: "Nitric Oxide and Microvessel Permeability in vivo".  

• NIH/NHLBI  RO1 (Pingnian He, Principal Investigator)
  Title: "Cellular Modulation of Microvessel Permeability in vivo".

• American Heart Association, Great River Affiliate Grant-In-Aid  (Pingnian He, Co-Investigator,  P.I Michel Miller)
  Title: "Lyme carditis: Borrelia burgdorferi interaction with microvessel in vivo".

Selected Publications:

1. Zeng, M, H. Zhang, C. Lowell, and P. He. Tumor necrosis factor-alpha-induced leukocyte adhesion and microvessel permeability. Am. J. Physiol. Heart and Circ Physiol 283: H2420-2430. Published on Aug. 8, 2002 as DOI 10.1152/ajpheart.00787.2001. PMID: 12388263 PubMed Article
2. Zhu, L, Schwegler-Berry D, Castranova V, and He P. Internalization of caveolin-1 scaffolding domain facilitated by Antennapedia homeodomain attenuates PAF-induced increase in microvessel permeability. Am J Physiol, Heart and Circ Physiol. 286: H195-H201. 2005 PMID: 12946927 PubMed Article
3. Bernatchez PN, Bauer PM, Yu J, Prendergast JS, He P, Sessa WC. Dissecting the molecular control of endothelial NO synthase by caveolin-1 using cell-permeable peptides. Proc Natl Acad Sci U S A. 102: 761-766. 2005 PMID: 15637154 PubMed Article
4. Minnear, F., L. Zhu, and P. He. Sphingosine 1-phosphate prevents platelet activating factor-induced increase in hydraulic conductivity in mesenteric venules: pertussis toxin-sensitive. Am J Physiol, Heart and Circ Physiol 289: H840-H844. 2005 PMID: 15778280 PubMed Article
5. Zhu, L and He P. Platelet activating factor increases endothelial [Ca2+]i and nitric oxide production in individually perfused intact microvessels. Am J Physiol, Heart and Circ Physiol 288: H2869-H2877. 2005 PMID: 15665052 PubMed Article
6. Zhu, L, Castranova V, and P He. FMLP-stimulated neutrophils increase endothelial [Ca2+]i and microvessel permeability in the absence of adhesion: Role of reactive oxygen species. Am. J. Physiol.Heart and Circ Physiol. 288: H1331-H1338. 2006 PMID: 15498822 PubMed Article
7. Zhu, L and He P. FMLP-Stimulated Release of Reactive Oxygen Species from TNF-a-Induced Adherent Leukocytes Increases Microvessel Permeability. Am J Physiol, Heart and Circ Physiol. 290: H365-H372. 2006 PMID: 16455097 PubMed Article
8. He, P, Zhang H, Zhu L. and Jiang Y, Zhou, X. Leukocyte/Platelet Aggregate Adhesion and Vascular Permeability: Role of Activated Endothelial Cells. Am J Physiol, Heart and Circ Physiol. 291: H591-H599. 2008 PMID: 16517944 PubMed Article
9. Jiang Y, Wen K, Zhou X, Schwegler-Berry D, Castranova V, He P. Three-dimensional localization and quantification of PAF-induced gap formation in intact venular microvessels. Am J Physiol, Heart and Circ Physiol. 295: H898-H906. 2008 PMID: 18515648 PubMed Article
10. Zhou X, Miller MR., Motaleb M, Charon N and He P. Borrelia burgdorferi directly increases permeability of individually perfused microvessels of rat mesentery via a plasmid-encoded mediator. Infection and Immunity. PLoS One, 3(12): e4101. doi:10.1371/journal.pone.0004101. 2009 PMID: 19116656 PubMed Article
11. Zhou X, Wen K, Yuan Dong; Ai L, and He P. Calcium influx-dependent differential actions of superoxide and hydrogen peroxide on microvessel permeability. Am J Physiol, Heart and Circ Physiol. 296: H1096-H1107, 2009. PMCID: PMC2670695 PubMed Article
12. He, P. Beyond tie-ing up endothelial adhesion: New insights into the action of Angiopoietin-1 in regulation of microvessel permeability. Cardiovascular Research, 83(1):H1-2, 2009. PubMed Article
13. Zhou X and He, P. Endothelial [Ca2+]i and caveolin-1 antagonistically regulate eNOS activity and microvessel permeability in rat venules. Cardiovascular Research doi:10.1093/cvr/cvq006 2010 PMID: 20080986 PubMed Article

Lab Personnel: Dong Yuan - Postdoc Sulei Xu - Ph.D. Student Xueping Zhou - Ph.D. Student Mingxia Wang - Research Scholar Christian Stork - Postdoc Jixiu Zhang - Research Scholar
	From left: Dong Yuan, Xueping Zhou, Mingxia Wang, Dr. Ping He, Sulei Xu, Christian Stork and Jixiu Zhang