Corpus cavernous smooth muscle cells are the main functional component of the corpus cavernosum penis, whose phenotypic modulation is the key initial step in the proliferation and migration of smooth muscle cells. Therefore, an insight into the mechanism of the phenotypic modulation of smooth muscle cells and its influencing factors is important for the prevention and management of penis erectile dysfunction. Smooth muscle cells are generally divided into contracting (differentiated) and composing (undifferentiated, proliferated or dedifferentiated) types. It is found that TGF-beta, transcription factor E2F1, BTEB2 and insulin may affect the phenotypic modulation of smooth muscle cells. This paper presents an overview of the progress in the researches on the phenotypic modulation of corpus cavernous smooth muscle cells and its influencing factors.
Cells, Cultured ; Humans ; Male ; Muscle, Smooth ; cytology ; Myocytes, Smooth Muscle ; classification ; cytology ; ultrastructure ; Penis ; Phenotype

OBJECTIVETo investigate an effective method to produce large numbers of pure corporal smooth muscle cells in vitro according to the requirement of study.

METHODSIn this study, we used the primary tissue culture technique to isolate and culture the corpus cavernosum smooth muscle cells (CCSM) from human males with normal erectile function and New Zealand white rabbits. The cells were identified in regard to morphological and growing characteristics via immunohistochemical methods (including alpha-smooth muscle actin, desmin, myocin and factor VIII related antigen), special dye techniques (including Masson and Van Gieson) and transmission electron microscope.

RESULTSCCSM were isolated and cultured successfully with high purity. Morphologically, the cells were spindle shaped and grow on top of each other, resembling a "hill and valley" in appearance. When characterized in immunohistochemistry, the cells were stained with alpha-smooth muscle actin, desmin and myocin, but not with anti-factor VIII, an endothelial marker.

CONCLUSIONThe CCSM, which can be isolated and cultured successfully, may be used for further studying their biological function. The CCSM cultured in vitro was proved to be useful to evaluate and investigate the effect of some new medicine for penile erection. There is also a clinical and theoretical significance in further studying the experimental mechanisms of erectile dysfunction.

Animals ; Cell Division ; Cells, Cultured ; Erectile Dysfunction ; etiology ; Humans ; Immunohistochemistry ; Male ; Myocytes, Smooth Muscle ; chemistry ; cytology ; ultrastructure ; Penis ; cytology ; Rabbits

OBJECTIVETo investigate the role of ICC-like cells in bladder neuromodulation in rat urinary bladder.

METHODS14 SD rats and 1 guinea pig were sacrificed in this study. The ultra structural relationships among interstitial cells, nerves and detrusor smooth muscle cells (DSMCs) of urinary bladder were investigated by transmission electron microscopy (TEM). c-kit immunofluorescence was used to identify ICC-like cells in SD rat urinary bladder and the structural relationship between ICC-like cells and nerve terminals was studied by immunofluorescence (double-label).

RESULTSGap junction between ICC-like cells and DSMCs was confirmed by TEM. ICC-like cells were very close apposition with nerve terminals under TEM. ICC-like cells were identified to exist in sub-urothelium layer, along the longitude of smooth muscle bundles and among detrusor smooth muscle in SD rat urinary bladder by c-kit immunofluorescence. Double-labeled tissue with c-kit and PGP9.5 antibodies also showed that ICC-like cells were very close apposition with nerve terminals in SD rat bladder.

CONCLUSIONSMorphological study indicated that ICC-like cells in rat urinary bladder may play an important role in detrusor neuromodulation. Further study on function will be helpful for elucidating the mechanism of bladder neuromodulation clearly.

Animals ; Cells, Cultured ; Female ; Gap Junctions ; Guinea Pigs ; Male ; Muscle, Smooth ; innervation ; Myocytes, Smooth Muscle ; cytology ; ultrastructure ; Nerve Endings ; ultrastructure ; Rats ; Rats, Sprague-Dawley ; Urinary Bladder ; cytology ; innervation ; ultrastructure

OBJECTIVEThe radial artery differs from internal mammary artery in its vascular biology and long-term patency after coronary artery bypass grafting (CABG). This study was designed to investigate their ultrastructural differences that may have implications in arterial remodeling and graft failure.

METHODSThirty-four radial artery and 11 internal mammary artery samples were obtained from patients underwent CABG, and subjected to routine electron microscopic examination. A semi-quantitative method was used to evaluate secretary endothelial cells, endothelial denudation, synthetic smooth muscle cells (SMCs), matrix accumulation, lipid deposition and medial submicroscopic calcification.

RESULTSCompared with internal mammary arteries, the radial arteries had more secretory endothelial cells (47.1%, 16/34 vs 27.2%, 3/ 11) and synthetic type SMCs in a background (14.4% vs 0.9%) that had more intimal lipid deposition and matrix accumulation (14.7%, 5/34 vs 9.1%, 1/11). Matrix vesicles and calcifications were frequently present in the media of both types of arteries. The calcifications, however, could not be visualized by routine histological stains, and therefore, named as submicroscopic calcification in this study. Fewer endothelial denudations were observed in the radial arteries, but no differences in medial lipid deposition and submicroscopic calcification were observed between these two types of arteries. The ultrastructural features and the arrangement of medial SMCs in radial arteries were similar to those of internal mammary arteries.

CONCLUSIONSRadial arteries have a higher SMC proliferative potential and more actively secretory status of endothelial cells, which may enhance the remodeling process and correlate with a decreased long-term patency. Better preservation of endothelial cells in radial arteries could be attributed to the "no touch" technique utilized in surgical harvesting. The significance of submicroscopic medial calcification during graft remodeling requires further investigations.

Calcinosis ; Coronary Artery Bypass ; methods ; Coronary Disease ; pathology ; surgery ; Endothelial Cells ; pathology ; ultrastructure ; Humans ; Male ; Mammary Arteries ; transplantation ; ultrastructure ; Microscopy, Electron ; Middle Aged ; Myocytes, Smooth Muscle ; pathology ; ultrastructure ; Radial Artery ; transplantation ; ultrastructure ; Tunica Intima ; pathology ; ultrastructure

BACKGROUNDAutophagy has been found to be involved in animal and cell models of atherosclerosis, but to date, it lacks general observation in human atherosclerotic plaques. Here, we investigated autophagy in smooth muscle cells (SMCs), endothelial cells (ECs), and macrophages in human atherosclerotic plaques via transmission electron microscopy (TEM), western blotting, and immunohistochemistry analysis.

METHODSThe histopathologic morphology of these plaques was observed via hematoxylin and eosin staining. The ultrastructural morphology of the SMCs, ECs, and macrophages in these plaques was observed via TEM. The localization of microtubule-associated protein 1 light chain 3 (MAP1-LC3), a relatively special maker of autophagy, in plaques was observed by double fluorescent immunochemistry and western blotting.

RESULTSAll of these human atherosclerotic plaques were considered advanced and unstable in histologically observation. By double fluorescent immunochemistry, the expression of LC3-II increased in the SMCs of the fibrous cap, the macrophages, and the microvascular ECs of the plaque shoulders. The protein level of LC3-II by western blotting significantly increased in plaques compared with normal controls. In addition, TEM observation of plaques revealed certain features of autophagy in SMCs, ECs, and macrophages including the formation of myelin figures, vacuolization, and the accumulation of inclusions in the cytosol. These results indicate that autophagy is activated in SMCs, ECs, and macrophages in human advanced atherosclerotic plaques.

CONCLUSIONSOur study is to demonstrate the existence of autophagy in human atherosclerotic plaques by different methods, which may contribute to the development of pharmacological approaches to stabilize vulnerable and rupture-prone lesions.

Atherosclerosis ; metabolism ; physiopathology ; Autophagy ; physiology ; Endothelial Cells ; pathology ; Humans ; In Vitro Techniques ; Microscopy, Electron, Transmission ; Microtubule-Associated Proteins ; metabolism ; Myocytes, Smooth Muscle ; pathology ; Plaque, Atherosclerotic ; metabolism ; physiopathology ; ultrastructure

OBJECTIVETo investigate the changes in the morphology and cytoskeleton (CSL) content of the CSL in the colonic smooth muscle cells (SMCs) of rats in early postscald stage, so as to elucidate the mechanism of dysfunction of gastrointestinal motility.

METHODSSeventy Wistar rats were randomly divided into normal control (n = 10, without scald) , and scald ( n = 60, with 10 cm x 7 cm wound inflicted on the back) groups. The colonic smooth muscle tissue of 10 normal rats and scalded rats were harvested at 1, 3, 6, 12, 24 and 48 postscald hours( PSH) and divided into two parts: one for histologic examination, and the other for the detection of CSL changes in the colonic smooth muscle tissue by flowcytometry method.

RESULTSThe electron microscope examination showed that the arrangement of cytoskeleton of SMC of the scalded rats during 1 to 3 PSH was disordered, and sparse, and the condensed area was uneven, with fragmentation. But the morphology and distribution of CSL gradually restored to normal state during 6 to 12 PSH, and it approached that of normal group at 48 PSH. The CSL content in the colonic smooth muscle tissue of scalded rats was obviously increased at 1 PSH (610+/-23) , decreased thereafter, evidently lower than that in control group at 3 PSH (92+/-17) , and then it started to increase at 12 PSH, exceeding the normal value at 24 PSH, and continued to rise until 48 PSH. There was significant difference in CSL content in the colonic smooth muscle tissue of the rats between scald and control group ( P < 0.05 or 0. 01).

CONCLUSIONChanges in the morphology and CSL content in the colonic smooth muscle tissue can be observed at early stage after a scald, which imply the kinetic balance between damage and repair in the body. In addition, changes in CSL content in the colonic smooth muscle tissue may be important factors in producing colonic dysfunction, damage of intestinal wall structure, and dynamic abnormalities of the colonic smooth muscle.

Actins ; metabolism ; Animals ; Burns ; metabolism ; pathology ; Colon ; cytology ; Cytoskeleton ; ultrastructure ; Disease Models, Animal ; Microtubules ; Myocytes, Smooth Muscle ; cytology ; metabolism ; Rats ; Rats, Wistar

PURPOSE: Clinical studies have reported a correlation between pelvic ischemia and voiding dysfunction in elderly men. The aim of this study was to identify and compare prostate structural modifications in cultured cells and in a rabbit model after exposure to hypoxia, oxidative stress, and chronic ischemia. MATERIALS AND METHODS: Cultured human prostate smooth muscle cells (SMCs), epithelial cells (ECs), and stromal cells (SCs) were incubated under normoxia, hypoxia, and oxidative stress conditions by use of a computerized oxycycler system. We developed a rabbit model of chronic prostate ischemia by creating aorto-iliac arterial atherosclerosis. Markers of oxidative stress were examined by using fluorometric analysis and enzyme immunoassay. Prostate structure was examined by using Masson's trichrome staining and transmission electron microscopy (TEM). RESULTS: Lipid peroxidation was found in SMCs exposed to hypoxia and in all cell types exposed to oxidative stress. We identified protein oxidation in ECs exposed to hypoxia and in all cell types exposed to oxidative stress. Markers indicating oxidative damage were present in chronically ischemic rabbit prostate tissue. These reactions were associated with DNA damage. Prostate ischemia resulted in epithelial atrophy, loss of smooth muscle, and diffuse fibrosis. TEM showed swollen mitochondria with degraded cristae, loss of membrane, loss of Golgi bodies, degenerated nerves, and disrupted cell-to-cell junctions. CONCLUSIONS: Human prostate cells exhibited differential reactions to hypoxia and oxidative stress with widespread DNA damage. Structural modifications in ischemic prostate tissue were similar to those in cells exposed to oxidative stress. Structural changes due to ischemia and oxidative stress may contribute to prostatic noncompliance in aging men.
Animals ; Anoxia/*complications ; Atherosclerosis/complications ; Biomarkers ; Cells, Cultured ; DNA Damage ; Disease Models, Animal ; Epithelial Cells/ultrastructure ; Fibrosis ; Humans ; Ischemia/*complications ; Lipid Peroxidation ; Male ; Myocytes, Smooth Muscle/ultrastructure ; Nerve Degeneration ; *Oxidative Stress ; Prostate/*anatomy & histology/*cytology ; Rabbits ; Stromal Cells/ultrastructure ; Urinary Bladder Neck Obstruction/complications

OBJECTIVETo study the effects of the divided functional recipes of Dahuang Zhechong pill( DHZCP) on atherosclerosis in rabbits.

METHODThe atherosclerotic model was established by the combination of hypercholesterol feeding and immune-injured endothelium in rabbits. Male New Zealand rabbits were randomly divided into nine groups: normal group, model group, Danshen group (0. 5 g x kg(-1) ), the low-dose(0. 5 g x kg(-1) ) and high-dose( 1.0 g - kg(-1) ) groups of the first divided recipe, the low-dose(0. 75 g x kg-' ) and high-dose(1. 5 g x kg(-1)) groups of the second divided recipe, the low-dose(0. 8 g x kg(-1) ) and high-dose( 1.6 g x kg(-1) ) groups of the third divided recipe. The effects of the divided functional recipes of DHZCP were observed in macropathology, histopathology and ultrastructure. Image analyzing system was used to determine atherosclerotic plaque area, intima thickness(IT) and intima-media thickness(IMT) in rabbit aorta.

RESULTThe divided functional recipes of DHZCP could significantly decreased the deposit of lipid and the atherosclerotic plaque area in aorta intima, relieve the histopathological changes of atherosclerosis, and inhibited the proliferation of vascular smooth muscle cells and collagen to reduce pachynsis of vascular intima. The divided functional recipes of DHZCP also reduced IT, IMT and IT/MT and reversed the contractive vascular remodeling.

CONCLUSIONThe divided functional recipes of DHZCP produce the different anti-atherosclerotic action, among which the first divided functional recipe exhibits more effective action.

Animals ; Aorta ; drug effects ; pathology ; ultrastructure ; Atherosclerosis ; pathology ; prevention & control ; Cell Proliferation ; drug effects ; Cockroaches ; chemistry ; Dose-Response Relationship, Drug ; Drug Combinations ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; therapeutic use ; Male ; Medicine, Chinese Traditional ; Microscopy, Electron ; Myocytes, Smooth Muscle ; drug effects ; pathology ; ultrastructure ; Plants, Medicinal ; chemistry ; Rabbits ; Random Allocation ; Rheum ; chemistry ; Tunica Intima ; drug effects ; pathology

OBJECTIVETo study the expression of nNOS and ultrastructural changes in the penile tissue of rats with prolactinoma-induced erectile dysfunction (ED).

METHODSWe established the model of prolactinoma in 20 male Westar rats by peritoneal injection of diethylstilbestrol (DES) and treated the control rats with normal saline (n = 10) or sterilized arachis oil (n = 10). After 8 weeks, we performed the apomorphine test and measured the weight of the pituitary gland and the levels of serum prolactin (PRL) and testosterone (T) to confirm the successful construction of the prolactinoma-induced ED model. Then we determined the expression of nNOS in the penile tissue by immunohistochemistry and examined the ultrastructural changes of the penile cavernosum under the transmission electron microscope.

RESULTSThe prolactinoma-induced ED model was successfully established in 15 rats. The weight of the pituitary gland was significantly increased in the rats treated with DES as compared with the normal saline and sterilized arachis oil controls ([46.7 ± 15.5] vs [11.7 ± 2.4] and [12.4 ± 2.3] mg, both P < 0.05). The level of serum PRL was markedly higher while that of T remarkably lower in the former than in the latter two groups ([1,744.9 ± 304.5] vs [11.5 ± 2.4] and [10.6 ± 1.9] ng/ml, both P < 0.0l; [1.54 ± 0.46] vs [3.11 ± 1.08] and [3.04 ± 1.11] ng/ml, both P < 0.05). The rate of penile erection was significantly reduced in the prolactinoma-induced ED model rats in comparison with the normal saline and arachis oil controls (16.7% vs 100% and 87.5%, both P < 0.05), and so was the expression of nNOS in the penile tissue (0.024 ± 0.011 vs 0.066 ± 0.019 and 0.058 ± 0.021, both P < 0.05). Transmission electron microscopy manifested significant ultrastructural changes in the endothelial and smooth muscle cells of the cavernous tissue in the prolactinoma-induced ED models.

CONCLUSIONThe ultrastructural changes of the penile cavernous tissue and the reduced expression of nNOS in penile tissue may be the most important mechanisms of prolactinoma-induced ED in rats.

Animals ; Apomorphine ; Carcinogens ; Diethylstilbestrol ; Erectile Dysfunction ; etiology ; Humans ; Male ; Myocytes, Smooth Muscle ; ultrastructure ; Nitric Oxide Synthase Type I ; metabolism ; Organ Size ; Penile Erection ; Penis ; enzymology ; ultrastructure ; Pituitary Neoplasms ; chemically induced ; complications ; Prolactin ; blood ; Prolactinoma ; chemically induced ; complications ; Rats ; Rats, Wistar ; Testosterone ; blood

BACKGROUNDAn important physiological feature of asthma is the phenotypic change of airway smooth muscle cells (ASMCs), but the precise mechanisms behind the ASMCs' change remains unknown. Our study assessed whether p21Ras can directly modulate the phenotype of ASMCs.

METHODSRat ASMCs were treated with FTP III, a highly specific p21Ras inhibitor. ASMCs were identified via immunocytochemistry. The ultrastructure of cells was observed by electron microscopy, and the expression of α-actin was evaluated by Western blotting analysis. The levels of IL-6 and RANTES were measured by enzyme linked immunosorbent assay (ELISA).

RESULTSIt was observed that ASMCs in asthma exhibited a proliferative/secretory phenotype and were larger, denser and had many pseudopods, as well as increased signs of secretory organelles. Additionally, the level of α-actin, a marker of ASMCs, was reduced in asthmatic ASMCs and the secretion of IL-6 and RANTES was increased. When FTP III was added to asthmatic ASMCs it induced a contractile phenotype, with increased α-actin levels and reduced secretion of IL-6 and RANTES.

CONCLUSIONSIt appears that p21Ras induces asthmatic ASMCs to a proliferative/secretory phenotype, but its inhibitor FTP III, can significantly reverse this phenotype. The role of p21Ras in the ASMCs may be a new target for asthma treatment.

Actins ; metabolism ; Animals ; Asthma ; metabolism ; Blotting, Western ; Cells, Cultured ; Chemokine CCL5 ; metabolism ; Enzyme-Linked Immunosorbent Assay ; Interleukin-6 ; metabolism ; Lung ; cytology ; Male ; Microscopy, Electron, Transmission ; Myocytes, Smooth Muscle ; drug effects ; metabolism ; ultrastructure ; Organophosphonates ; pharmacology ; Proto-Oncogene Proteins p21(ras) ; antagonists & inhibitors ; metabolism ; Rats ; Rats, Sprague-Dawley ; Trachea ; cytology