In this paper, 2 cases of collagen Type Ⅲ glomerulopathy were analyzed. The clinical manifestations mainly included nephrotic syndrome, proteinuria, hypertension and renal dysfunction. One patient showed that the complement factor H-related protein 5 (CFHR5) gene was likely a disease-causing mutation. The pathological examination of renal tissues showed hyperplasia of mesangial matrix, sub-endothelial insertion, and double-track formation. Immunohistochemistry of Type III collagen was positive. Electron microscopy revealed that massive collagen fibers (40-70 nm in diameter) deposited in the mesangial matrix and basement membrane. As for the follow-up results, the normal renal function had kept steady and the proteinuria was moderate in 1 case treated with angiotensin Ⅱ receptor blocker. Due to other system disease, another case developed into acute kidney injury and then received hemodialysis. The clinical manifestations of collagen Type Ⅲ glomerulopathy was atypical, the light microscope pathological features were various, and the disease was mainly diagnosed by electron microscopy and immunohistochemistry.
Collagen Type III ; genetics ; Glomerular Mesangium ; Humans ; Kidney Diseases ; Kidney Glomerulus ; Proteinuria

OBJECTIVEThe aim of this study was to investigate the expression of collagen in the process of masseter muscle reattachment to the cortical and cancellous bones of mandible.

METHODSA total of nine adult goats were used in the study. One was the control. The other eight were treated with bilateral detachment of the masseter muscles. The biopsies of bone and muscle were taken at 2, 4, 8 and 12 weeks after the operation. The characteristics of the healing muscle-bone interfaces were examined using immunohistochemical techniques.

RESULTSImmunohistochemical analysis illustrated that the locations of collagen type I, II and III were different during the healing process, but similar in the cortical and cancellous bones.

CONCLUSIONThis study demonstrates that the distribution of the three types of collagens at the muscle-bone interfaces is associated with time, but not related with their locations.

Animals ; Collagen ; biosynthesis ; genetics ; Collagen Type I ; biosynthesis ; genetics ; Collagen Type II ; biosynthesis ; genetics ; Collagen Type III ; biosynthesis ; genetics ; Female ; Goats ; Male ; Mandible ; metabolism ; pathology ; surgery ; Masseter Muscle ; metabolism ; pathology ; surgery ; Wound Healing ; physiology

OBJECTIVETo investigate the feasibility of transfection of recombinant human endothelial nitric oxide synthase (eNOS) into human hypertrophic scar fibroblasts (HSFbs), and to observe NO secretion and the synthesis of collagen I and III.

METHODSRecombinant human eNOS with karyocyte expressive vector was constructed in vitro, then was transfected into HSFbs which was isolated from hypertrophic scar tissues and cultured in vitro (T group). The HSFbs untransfected (normal culture) or transfected with empty-vector was used as control group and empty-vector group respectively. The mRNA expression of eNOS, collagen I and III was determined by Realtime PCR. The content of NO was determined by NO assay kit.

RESULTSThe expression of eNOS mRNA in T group was 5.92 +/- 0.21, which was obviously higher than that in empty-vector group (0.98 +/- 0.13, P < 0.05). The expression of collagen I mRNA (0.76 +/- 0.15), and collagen III (0.79 +/- 0.08) in T group was significantly lower than those in empty-vector group (0.98 +/- 0.15, 1.02 +/- 0.12, P < 0.05, respectively). The content of NO in T group (36.1 +/- 0.8 micromol/L) was obviously higher than that in empty-vector group (28.4 +/- 1.0 micromol/L, P < 0.01) and control group (27.7 +/- 1.3 micromol/L, P < 0.01).

CONCLUSIONHSFbs can be the target cells for eNOS gene transfection. The transfected cells can express eNOS and produce NO, which inhibit the synthesis of collagen.

Cicatrix, Hypertrophic ; metabolism ; Collagen Type I ; genetics ; metabolism ; Collagen Type III ; metabolism ; Fibroblasts ; metabolism ; Humans ; In Vitro Techniques ; Nitric Oxide ; metabolism ; Nitric Oxide Synthase Type III ; genetics ; RNA, Messenger ; metabolism ; Transfection

Thoracic aortic dissection (TAD) without familial clustering or syndromic features is known as sporadic TAD (STAD). So far, the genetic basis of STAD remains unknown. Whole exome sequencing was performed in 223 STAD patients and 414 healthy controls from the Chinese Han population (N = 637). After population structure and genetic relationship and ancestry analyses, we used the optimal sequence kernel association test to identify the candidate genes or variants of STAD. We found that COL3A1 was significantly relevant to STAD (P = 7.35 × 10
Aneurysm, Dissecting/genetics* ; Case-Control Studies ; Cluster Analysis ; Cohort Studies ; Collagen Type III/genetics* ; Computational Biology ; Genetic Predisposition to Disease ; Humans

AIMTo observe the effects of hypoxia on DNA synthesis and the expression of collagen type I and III mRNA in cultured adult rat cardiac fibroblasts.

METHODSCardiac fibroblasts(CFs) were isolated from adult Wistar rat ventricule and cultured in vitro either in normoxic or hypoxic condition. Studies were conducted with the second passage of CFs. The changes of DNA synthesis was determined by measuring the incorporation of 3H-TdR into DNA and the changes of expression of pro-alpha1 (I) collagen, pro-alpha1(III) mRNA were measured by in situ hybridization respectively.

RESULTSThe 3H-TdR incorporation of CFs was increased by 34% (P < 0.05) and 36% (P < 0.01) after 6 h, 12 h hypoxia (2% O2) exposure respectively. The level of pro-alpha1(I) collagen mRNA expression was significantly elevated in the cells under hypoxia for 4 h, 8 h, and 12 h. The expression of pro-alpha1(III) mRNA increased when cells were cultured under hypoxia for 2 h.

CONCLUSIONThese results suggest that hypoxia alone can upregulate DNA synthesis and expression of collagen type I and III mRNA in adult rat cardiac fibroblasts. It may be one of the important mechanisms by which hypoxic myocardial fibrosis occur.

Animals ; Cell Hypoxia ; Cells, Cultured ; Collagen Type I ; metabolism ; Collagen Type III ; metabolism ; DNA ; biosynthesis ; Fibroblasts ; metabolism ; Male ; Myocytes, Cardiac ; cytology ; RNA, Messenger ; genetics ; Rats ; Rats, Wistar

OBJECTIVETo observe the in vitro effects of gamma-interferon (IFNgamma) on gene expression of collagen I (Col I), III (Col III) and tissue inhibitor of metalloprotenase 1 (TIMP1) of HSC-T6 cells.

METHODSCultured HSC-T6 cells were exposed to IFNgamma at concentrations of 0.1, 1, 10, 10(2), 10(3), 10(4), 2.5 x 10(5), 5 x10(5) U/ml for 48 hours. 4,5-simethylthiazaoly colormetric assay was used to evaluate the effect of IFNgamma on HSC-T6 cell proliferation. After incubating with IFNgamma (1 U/ml, 10(2) U/ml and 10(4) U/ml) for 48 hours, HSC-T6 cells were harvested to detect Col I, Col III and TIMP1 steady state mRNA levels by quantitative reverse-transcription polymerase chain reaction (RT-PCR).

RESULTSThe Col I, Col III and TIMP1 mRNA levels of the control group were 2.86+/-0.21, 2.00+/-0.23 and 3.90+/-0.81, respectively. Col I and Col III mRNA levels in HSC-T6 cells treated by different concentrations of IFNgamma were lower than that of the controls (P < 0.01). There was no significant difference in TIMP1 mRNA levels between IFNgamma groups and controls.

CONCLUSIONIFNgamma suppresses expression of Col I and Col III whereas it has no effect on TIMP1 mRNA expression. The antifibrotic mechanism of IFNgamma may be partly due to its down-regulation of Col I and Col III mRNA levels in HSC-T6 cells.

Animals ; Cell Line ; Collagen Type I ; biosynthesis ; genetics ; Collagen Type III ; biosynthesis ; genetics ; Hepatocytes ; cytology ; metabolism ; Interferon-gamma ; pharmacology ; RNA, Messenger ; biosynthesis ; genetics ; Rats ; Rats, Sprague-Dawley ; Tissue Inhibitor of Metalloproteinase-1 ; biosynthesis ; genetics

BACKGROUNDThis study was designed to investigate changes in mRNA levels of transforming growth factor-beta (TGF-beta), collagen I, and collagen III in autogenous vein grafts.

METHODSTwenty-four New Zealand rabbits were randomly divided into 4 groups with 6 rabbits each. The external jugular veins of the New Zealand rabbits were harvested and grafted into the ipsilateral carotid artery. All rabbits were fed with a standard diet. After the operation, the rabbits were sacrificed at 1, 2, 3, or 4 weeks. TGF-beta, collagen I, and collagen III mRNA levels in the venous grafts were measured by semiquantitative methods at every time point. The contralateral external jugular veins were also harvested and analyzed as controls. Glyceraldehyde-3-phosphate dehydrogenase was used as an internal standard to normalize all samples for potential variations in mRNA content. In order to observe the expression of TGF-beta protein, immunohistochemical SABC methods were used.

RESULTSOne week postoperation, the mRNA level of TGF-beta was upregulated to 1.73 +/- 0.19 in the vein graft and 1.21 +/- 0.16 in the control vein (P < 0.01). High mRNA levels were maintained until week 4 postoperation. The mRNA levels of collagen I and collagen III were also significantly increased to 2.18 +/- 0.21 versus 1.12 +/- 0.24 and 1.08 +/- 0.13 versus 0.83 +/- 0.12, respectively (P < 0.05). Immunohistochemical staining revealed a higher density of TGF-beta expression in the vein grafts.

CONCLUSIONSAn uninterrupted increase in mRNA levels of TGF-beta, collagen I, and collagen III is observed in autogenous vein grafts. This increase may be the major cause of intimal hyperplasia, sclerosis, and even graft failure.

Animals ; Collagen Type I ; genetics ; Collagen Type III ; genetics ; Female ; Immunohistochemistry ; Jugular Veins ; transplantation ; Male ; RNA, Messenger ; analysis ; Rabbits ; Reverse Transcriptase Polymerase Chain Reaction ; Transforming Growth Factor beta ; analysis ; genetics ; Transplantation, Autologous

OBJECTIVETo suppress COL1A1 and COL3A1 gene expressions in human skin fibroblasts (HSFs) by means of RNA interference (RNAi).

METHODSThree small interfering RNA (siRNA) expression cassette (SEC) sequences were designed for each of the COL1A1 and COL3A1 gene sequences available in GenBank. The synthesized SECs capable of effective gene suppression were transfected into cultured HSFs, either after cloning into the expression vector or mediated by Lipofectamine 2000, and the suppression of the target genes at both mRNA and protein levels was determined by quantitative fluorescence RT-PCR and Western blotting, respectively.

RESULTSTransfection of the SECs into HSFs resulted in specific depression of COL1A1 and COL3A1 expressions (down to 5.00% and 6.48%, respectively). The expression vector-mediated RNAi established a HSF cell line with persistent gene knockdown for over 30 days (to 25.21% and 22.12%, respectively).

CONCLUSIONCOL1A1 and COL3A1 gene expressions can be specifically and efficiently inhibited in HSFs by either liposome- or vector-mediated SEC transfection.

Blotting, Western ; Cells, Cultured ; Collagen Type I ; biosynthesis ; genetics ; Collagen Type III ; biosynthesis ; genetics ; Fibroblasts ; cytology ; metabolism ; Humans ; RNA Interference ; RNA, Small Interfering ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Skin ; cytology ; Transfection ; methods

OBJECTIVESTo study the effects of Sanqi qisodium hyaluronate gel on collagen-I and collagen-III expression in the process of rabbits' epidural scar formation after operation.

METHODSNinety-six white rabbits with 6-month-old, half males and half females, weighted from 2 to 2.5 kg, which were randomly divided into normal saline group (A), Sanqi group (B), qisodium hyaluronate group (C) and Sanqi qisodium hyaluronate gel group (D). The laminectomy of rabbits were performed in group A, B, C, D, the duras were surrounded with normal saline, Sanqi liquid, qisodium hyaluronate and Sanqi qisodium hyaluronate gel respectively. Animals of each group were killed at 1, 2, 4, 8 weeks after operation. Use Masson staining for histological observation of collagen, and in situ hybridization staining for the analysis of collagen-I and collagen-III expression.

RESULTSIn the Masson staining, Sanqi qisodium hyaluronate gel group was more regular than the control group in the shape of collagen texture. As to the expression of collagen-I, and Sanqi qisodium hyaluronate gel group was lower than normal saline group, the Sanqi group and qisodium hyaluronate group at 4 weeks after using medicine (P < 0.01); while the Sanqi qisodium hyaluronate gel group was higher than normal saline group, Sanqi group and qisodium hyaluronate group in the collagen-III expression (P < 0.01).

CONCLUSIONSSanqi qisodium hyaluronate gel could improve collagen's arrangement of the rabbit's epidural scar after operation, reduce its rigidity and increase flexibility.

Animals ; Cicatrix ; etiology ; metabolism ; Collagen Type I ; genetics ; metabolism ; Collagen Type III ; genetics ; metabolism ; Epidural Space ; Female ; Gels ; Gene Expression Regulation ; drug effects ; Hyaluronic Acid ; chemistry ; Laminectomy ; adverse effects ; Male ; RNA, Messenger ; genetics ; metabolism ; Rabbits

OBJECTIVETo investigate the effect of leptin on collagen systhesis in wounded rats.

METHODSThirty male Wistar rats, weight (180 +/- 20)g, were randomly divided into three groups (n = 10) by weight: normal depilation group, wound control group and leptin treatment group and ten rats were included in each group. A full-thickness defect measuring 2 x 2.5 cm was made in the back of rats in wound control group and leptin treatment group. Each wound in rats of leptin treatment group was applied topically with 0.1 ml leptin solution (2.0 microg leptin), daily for 7 days and that of wound control group with equivalent saline solution. All rats were killed and then granulation tissues samples and skin were collected to examine the synthesis of collagen.

RESULTSHydroxyproline content in granulation tissues of in leptin treatment group (33.92 +/- 3.09) mg/g were significantly increased than those in control group (29.55 +/- 3.59 mg/g, P < 0.05). The mRNA expressions of collagen I and III were significantly enhanced in leptin treatment group (0.96 +/- 0.09, 0.09 +/- 0.06) than those in control group (0.80 +/- 0.03, 0.08 +/- 0.03). The levels of type I and III collagen were significantly increased in leptin treatment group than those in control group.

CONCLUSIONLeptin applied topically can accelerate wound healing through enhancing gene expression of type I and III collagen and synthesis of collagen in wound tissue.

Administration, Cutaneous ; Animals ; Collagen Type I ; genetics ; metabolism ; Collagen Type III ; genetics ; metabolism ; Leptin ; administration & dosage ; therapeutic use ; Male ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Wistar ; Wound Healing ; drug effects ; Wounds and Injuries ; drug therapy