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Objective::To investigate the protective effect of Ginseng Radix et Rhizoma, Notoginseng Radix et Rhizoma and Chuanxiong Rhizoma (GNC) extracts on myocardial fibrosis in diabetic mice by observing the degree of myocardial fibrosis and collagen types I (Collagen Ⅰ), collagen types Ⅲ (Collagen Ⅲ) and transforming growth factor-β1 (TGF-β1) protein expression in myocardial tissues. Method::A diabetic mice model was induced by streptozotocin (STZ) and high-fat diet. A normal control group was established. According to random number table method, diabetic mice were divided into model group, GNC low-dose and high-dose groups (0.819, 1.638 g·kg-1), and metformin group (150 mg·kg-1). Intragastrical administration was given in all groups, and the mice in normal control group received an equal dose of deionized water once a day for 9 weeks. The myocardial interstitial fibrosis in mice was observed by Masson trichromatic staining. Image-pro plus 6.0 analysis software was used to calculate the ratio of collagen area to total area. Immunohistochemistry was used to detect Collagen I, Collagen Ⅲ and TGF-β1 protein expression in myocardial tissues. The protein expression electrophoresis and gray value levels of Collagen I, Collagen Ⅲ and TGF-β1 in the myocardial tissues were detected by Western blot. Result::The results of Masson staining showed that as compared with the normal control group, the myocardial cells of diabetic mice were hypertrophic and disordered, and the myocardial stroma, especially the blue-stained collagenous fibers around the blood vessels, were heavily deposited and connected to each other in a network (P<0.01). As compared with the model group, the arrangement of myocardial cells was significantly improved in GNC low-dose and high-dose groups and metformin group, and the collagenous fibers in the myocardial stroma were significantly decreased (P<0.05). Immunohistochemistry and Western blot results showed positive expression of Collagen Ⅰ, Collagen Ⅲ and TGF-β1 in myocardial tissues, with significantly increased content of protein expression in diabetic mice (P<0.05, P<0.01). As compared with the model group, the positive protein expression decreased and the protein content tended to be normal in each administration group (P<0.05, P<0.01). Conclusion::High-fat diet combined with STZ can induce myocardial fibrosis in diabetic mice, and increase Collagen I, Collagen Ⅲ and TGF-β1 protein expression. Ginseng Radix et Rhizoma, Notoginseng Radix et Rhizoma and Chuanxiong Rhizoma extracts can improve myocardial fibrosis in diabetic mice by regulating the expression of Collagen I, Collagen Ⅲ and TGF-β1 protein.
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OBJECTIVE@#To verify the effect of the mutant gene vps4b on the expression of tooth development-related proteins, dentin sialophosphoprotein (DSPP) and collagenⅠ (COL-Ⅰ).@*METHODS@#Paraffin tissue sections of the first molar tooth germ were obtained from the heads of fetal mice at the embryonic stages of 13.5, 14.5, and 16.5 days and from the mandibles of larvae aged 2.5 and 7 days after birth. The immunohistochemical method was used to detect the expression and location of DSPP and COL-Ⅰ in wild-type mouse and vps4b knockout mouse.@*RESULTS@#DSPP and COL-Ⅰ were not found in the bud and cap stages of wild-type mouse molar germ. In the bell stage, DSPP was positively expressed in the inner enamel epithelium and dental papilla, whereas COL-Ⅰ was strongly expressed in the dental papilla and dental follicle. During the secretory and mineralized periods, DSPP and COL-Ⅰ were intensely observed in ameloblasts, odontoblasts, and dental follicles, but COL-Ⅰ was also expressed in the dental papilla. After vps4b gene knockout, DSPP was not expressed in the dental papilla of the bell stage and in the dental papilla and dental follicle of the secretory phase. The expression position of COL-Ⅰ in the bell and mineralization phase was consistent with that in the wild-type mice. Moreover, the expression of COL-Ⅰ in the dental papilla changed in the secretory stage.@*CONCLUSIONS@#Gene vps4b plays a significant role in the development of tooth germ. The expression of DSPP and COL-Ⅰ may be controlled by gene vps4b and regulates the development of tooth dentin and cementum together with vps4b.
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Animals , Mice , ATPases Associated with Diverse Cellular Activities , Genetics , Collagen , Metabolism , Endosomal Sorting Complexes Required for Transport , Genetics , Extracellular Matrix Proteins , Metabolism , Mice, Knockout , Molar , Odontoblasts , Phosphoproteins , Metabolism , Sialoglycoproteins , Metabolism , Tooth GermABSTRACT
Objective: To study the effect of Yangxinkang tablets on myocardial fibrosis in mice after heart failure, and to explore its mechanism.Method: The model of chronic heart failure in mice was established by thoracic aorta constriction (TAC). After successful modeling, mice were randomly divided into sham operation group, model group, 3-methyladenine(3-MA,15 mg·kg-1) autophagy inhibitor group, Yangxinkang tablets high, medium, and low dose groups (1 170,585,390 mg·kg-1).The sham operation group received equal volume of distilled water. After 30 days, cardiac ultrasound was performed to collect hemodynamic parameters. Cardiac paraffin slices were stained with Masson to observe the morphological changes and fibrosis of cardiomyocytes. Western blot was used to detect lysosome-associated membrane protein(LAMP), microtubule-associated protein light chain 3 (LC3), Beclin-1 autophagyportein, α-smooth muscle activin(α-SMA),Collagen Ⅰ,Collagen Ⅲ protein expression.Result: As compared with normal group, the left ventricular ejection fraction (LVEF) and fractional shortening(FS) were significantly decreased(PPPα-SMA, Collagen Ⅰ, Collagen Ⅲ, LAMP, LC3, and Beclin-1 were significantly increased in model group (PPPα-SMA,Collagen Ⅰ,Collagen Ⅲ,LAMP,LC3 and Beclin-1 were decreased in 3-MA group, Yangxinkang high and medium dose groups(PConclusion: Yangxinkang tablets can reduce myocardial fibrosis and improve cardiac function in mice with heart failure probably by down-regulating autophagy.
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Objective: To study the protective effect and mechanism of compound Danshen dropping pills on the heart of rats with type 2 diabetic cardiomyopathy. Method: Seventy-five male SD rats were randomly divided into blank group (15 rats) and model group (60 rats). The model group was fed with high-sugar and high-fat diet, high-sugar and high-fat milk and sucrose water. Six weeks later, rats with foodborne obesity (body mass ≥ the mean body mass of blank control group + 2 times standard deviation) were selected and intraperitoneally injected with 1% streptozotocin (STZ)28 mg·kg-1. After 12 weeks, rats with fasting blood glucose ≥ 11.1 mmol·L-1 and polydipsia, diuresis, polyphagia and weight loss were randomly divided into model group, compound Danshen dropping pills group (0.5 g·kg-1) and metformin group (0.5 g·kg-1) and intervened for 12 weeks. At the end of the experiment, echocardiography was used to evaluate cardiac function. Blood samples were taken to determine the corresponding biochemical indicators of rats. Histopathological changes of myocardium were observed by hematoxylin-eosin (HE) and Masson staining. The ultrastructure of myocardium was observed under electron microscope. Western blot was used to detect the expressions of vascular endothelial growth factor (VEGF), transforming growth factor-beta 1 (TGF-β1) and type Ⅰ collagen (Collagen Ⅰ). Real-time fluorescence quantitative analysis (Real-time PCR) was used to detect the expression of miRNA-200b. Result: Compared with the blank group, the heart weight index (HMI), fasting blood glucose (FBG), glycosylated hemoglobin (HbA1c), fasting insulin (FINS), total cholesterol (TC), triglyceride (TG), low-density lipoprotein cholesterin(LDL-C) levels in the model group were significantly increased (PPPPPβ1 and Collagen I increased (PPPPPPβ1 and Collagen I was decreased (PPConclusion: Compound Danshen dropping pills may play a protective role in the heart of type 2 diabetic cardiomyopathy rats by lowering lipid, up-regulating the expression of miR-200b and inhibiting the expression levels of VEGF, TGF-β1 and Collagen I.
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Objective: To investigate the effect of Kangxianling decoction on renal function and expression of extracellular matrix(ECM) in renal tissue of rats with renal fibrosis induced by 5/6 nephrectomy. Method: Totally 50 SD rats were randomly divided into control group (n=7), sham-operation group (n=7), operation group (n=36). The 5/6 nephrectomized renal fibrosis model was established in operation group. After successful modeling, rats were randomly divided into model group, Kangxianling group and losartan potassium group. The losartan potassium group and the Kangxianling group were given losartan potassium (33.3 g·kg-1) and Kangxianling decoction (21 g·kg-1) every day. The control group, sham-operation group and model group were all treated with equal volume of normal saline. Rats were put to death after 24 weeks of consecutive medication, and renal function, 24 h urine volume and 24 h-Pro quantitation of each group were measured. The expressions of collagen Ⅰ(ColⅠ) and ColⅢ in renal tissues were determined by immunohistochemistry and Western blot. Result: Compared with the normal group and sham-operation group, serum creatinine(SCr), blood urea nitrogen(BUN), 24 h urine volume, 24 h-Pro quantitation, renal tissue ColⅠ and ColⅢ expression levels were significantly increased in the model group (PPPConclusion: Kangxianling decoction can down-regulate the expressions of ColⅠ and ColⅢ in kidney tissue, there by inhibiting the accumulation of ECM, and exerting the effect in delaying renal fibrosis and protecting renal function.
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OBJECTIVE: To observe the effect of electroacupuncture (EA) at "Weizhong" (BL40) on histopathological changes and expression of extracellular matrix (ECM) component Collagen Ⅰ， matrix metalloproteinases 2 (MMP2), MyoD and Pax7 proteins of lumbar muscle tissues in rats with lumbar multifidus muscle injury (LMMI), so as to explore its underlying mechanisms in improving muscular injury. METHODS: A total of 24 male SD rats were equally randomized into blank control, model and EA groups. The LMMI model was established by injection of 0.5% Bupivacaine (100 µL/ point) into bilateral multifidus muscles of lumbar 4 and 5 (4 points). EA (2 Hz/100 Hz in frequency, 1－2 mA) was applied to BL40 for 20 min, once a day for 3 days. The morphological changes of the left lumbar multifidus muscle were observed under microscope after H．E. and Masson staining, and the expression of Collagen Ⅰ， MMP2, MyoD and Pax7 of the right lumbar multifidus muscle was determined by Western blot. RESULTS: H．E. staining showed large areas of degeneration and necrosis of muscle fibers, and vacuolar structure formed by degradation of muscle fibers in the model group, and newborn juvenile muscle fibers with different diameters in the EA group. Masson staining showed a large number of morphologically damaged muscle fibers and blue stained collagen fibers in the model group, and significantly reduced collagen fibers as well as new muscle fibers with uneven diameters in the EA group. The expression levels of Collagen Ⅰ， MMP2 and MyoD proteins were significantly up-regulated (P<0.01, P<0.05), and that of Pax7 was considerably down-regulated in the model group relative to the control group (P<0.01). After EA intervention, the expression levels of Collagen Ⅰ was significantly down-regulated (P<0.01), and those of MMP2, MyoD and Pax7 proteins were obviously or further obviously up-regulated in the EA group compared with the model group (P<0.01, P<0.05). CONCLUSION: EA at BL40 can reduce the degree of skeletal muscle fibrosis to promote the regeneration of the injured multifidus at the early phase, which may be related to its effect in regulating the expression of Collagen Ⅰ and MMP2 proteins.
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AIM To investigate the effect and mechanism of methanolic extract of Eupatorium (MEOE) to model rats with chronic soft tissue injury.METHODS The model rats were established by mechanical injury and a subsequent two-week normal feeding for respective administration of high,medium and small dosage of MEOE once a day successively for 14 days.An array of indices,the level of superoxide dismutase (SOD),malondialdehyde (MDA),prostaglandin E2 (PGE2),nitric oxide (NO),interleukin-6 (IL-6) and histamine,the expression of tumor necrosis factor-alpha (TNF-α),nitric oxide (NO) and Collagen-Ⅰ/Ⅲ,and the activity of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) were measured to analyze the effect of MEOE to model rats with chronic muscle injury.RESULTS MEOE resulted in apparent reduction of contents of MDA,PGE2 and NO,and the levels of TNF-α and IL-6 in muscular tissue (P ＜ 0.05),significantly increased of the SOD in muscular tissue (P ＜ 0.01),a remarkably inhibited expression of the tissue Collagen-Ⅰ/Ⅲ protein (P ＜ 0.01),and significantly improved activity of tissue VEGF and bFGF (P ＜ 0.01).CONCLUSION The certain therapeutic effects of MEOE to rats with chronic muscle injury may correlate with its influence to the levels of inflammatory factors inhibition,the oxidative stress relief,the overexpression of collagen-Ⅰ/Ⅲ inhibition,the VEGF and bFGF activity improvement,and the time spare from the repairing.
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Aim To explore the inhibitory effect of Euonymus alatus on hepatic fibrosis induced by carbon tetrachloride (CCl4) in mice and its mechanism. Methods Eighty C57BL/6 male mice were randomly divided into eight groups: normal group, CCl4model group, Euonymus alatus(EA) ethanol extracts groups in early stage(EAE), EA ethanol extracts groups in later stage(EAL),two drug groups with low/medium/high dose(EAE-L/M/H, EAL-L/M/H), with 10 mice in each group. Fibrosis model was established by injecting CCl4in peritoneal cavity,and the study lasted for 30 days. Different doses of drugs were given from 1 st day to 15 th day in EAE while from 16 th day to 30 th day in EAL,then all mice were sacrificed to for the observation of the morphological changes and collage-nous fiber by HE and Masson staining. Liver index, ALT,AST and TNF-α were tested by ELISA. The ex-pressions of α-SMA and CollagenⅠwere measured by immunohistochemistry and Western blot. Results Compared to normal group, liver index, ALT, AST, TNF-α, α-SMA and CollagenⅠ in EA groups were lower than those in model group in a dose-dependent manner(P<0.01 or P<0.05). Liver morphology and collagenous fiber in EAE and EAL were better than those in model group in a dose-dependent manner. The effect of EAE were superior to that of the EAL in HE, Masson, α-SMA, Collagen Ⅰ indexes(P <0.05). Conclusions Euonymus alatus may inhibit the process of hepatic fibrosis in mice with dose-effect de-pendence, and drug treatment in early stage performs better,which may be related to the decrease of TNF-α that affects the expression of α-SMA and Collagen Ⅰ.
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Objective To study the effect of S1P on HLF cell fibrosis and its mechanism. Methods (1) The expression of ECM in HLF cells was analyzed by using Western Blot after treatment by S1P(1 μmol/L), FTY720-P(5μmol/L),ponesimod(5μmol/L)and SEW2871(5μmol/L)24 h;(2)The HLF cells were pre-treated using selective S1PR antagonist W146(1 μmol/L),JTE-013(0.2 μmol/L),and TY-52156(1.25 μmol/L)1 h before incubation by S1P and S1PR agonists 24 h and then the expression of ECM was analyzed;(3)The HLF cells were pre-incubated using JTE-013(0.2μmol/L)and TY-52156(1.25μmol/L)for 1 h and then the expression of ECM was analyzedafter being treated by S1P and S1PR agonists 24 h. Results (1)S1P and selective S1P receptor agonist increased the expression of ECM to various extents;(2)The S1P1R antagonist W146 did not affectthe expression of ECM induced by S1P and S1PR agonists and S1P2R antagonist JTE-013 and S1P3R antagonist TY-52156 both decreased the expression of ECM induced by S1P and S1PR agonists;(3)The expression of ECM induced by S1P and S1PR agonists further decreased using both JTE-013 and TY-52156 but not using ponesimod. Conclusion S1P2R and S1P3R are activated under the influence of S1P so as to increase the synthesis of ECM and promote fibrosis gene expression in HLF cells.
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Objective To explore the effect of TLR2 on collagen Ⅰ(colⅠ) in adipose tissue of high-fat-diet induced obese mice.Methods Male C57bl/6J mice and TLR2 knockout mice were divided into groups according to high fat diet or normal chow.Total collagen, TLR2, colⅠ, MMP1, MMP2, TIMP1, colⅠα1 mRNA and colⅠα2 mRNA in adipose tissue were measured at the end of the experiments.Results Total collagen, TLR2, colⅠ, MMP2, TIMP1, colⅠα1 mRNA, and colⅠα2 mRNA in adipose tissue increased while MMP1 in adipose tissue decreased in mice with high fat diet.Decreased levels of total collagen, colⅠ, MMP2, TIMP1, colⅠα1 mRNA and colⅠα2 mRNA in adipose tissue were detected in TLR2 gene knockout mice with high fat diet.However, there was an increased level of MMP1 in TLR2 gene knockout mice with high fat diet.Conclusion In high-fat-diet induced obese mice, deposition of colⅠ in adipose tissue seems to be alleviated by TLR2 gene knockout via MMP1 and TIMP1.
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Aim To investigate the effect of thioester-ase superfamily member 4(THEM4)expression on col-lagen secretion in human renal proximal tubular epithe-lial cells (HKC)treated with high glucose. Methods In order to examine the direct effect of THEM4 ex-pression vector on PI3K/ Akt pathway and collagen se-cretion,pYr-ads-4-THEM4 expression vector was con-structed and transfected into the HKC with lipo-fectamine 2000 in vitro. HKC cells were randomly di-vided into four groups:normal glucose group (Con-trol),high glucose group (HG),high glucose plus pYr-ads-4-THEM4 vector group (HG + THEM4 vec-tor) and high glucose plus pYr-adshuttle-4 vector group (HG + V vector). After 48 h with HG stimula-tion,the cells were collected for extraction of protein and phospho-Akt (Ser 473),THEM4,TGF-β1 andα-SMA protein expression were examined by Western blot and immunofluorescence staining respectively. Col Ⅰ and Col Ⅲ were detected using the competitive sandwich ELISA kit according to the manufacturer's instructions. Results High glucose inhibited THEM4 expression,and induced increased phospho-Akt (Ser 473),TGF-β1,α-SMA and secreted ColⅠand secre-ted Col Ⅲ in HKC cells. Up-regulation of THEM4 re-versed high glucose-induced decreased THEM4,in-creased phospho-Akt (Ser 473),TGF-β1,α-SMA, secreted Col Ⅰ and secreted Col Ⅲ in HKC cells. Conclusion The up-regulation of THEM4 may de-crease Col Ⅰ and Col Ⅲ secretion by inhibiting the phosphorylation of Akt and down-regulating the expres-sion of TGF-β1 and α-SMA in high glucose-induced HKC cells.
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AIM:To explore the effects of chloroquine (CQ) on collagen Ⅰ and collagen Ⅲ expression in activated rat hepatic stellate cell line HSC-T6 and the possible mechanism.METHODS:Transforming growth factor-β1 (TGF-β1) was used to activate HSC-T6 cells and 3 doses of CQ was administered for 24 h.The cells were divided into 5 groups as follows:control group,TGF-β1 group,TGF-β1 + CQ (15 μmol/L) group,TGF-β1 + CQ (30 μ mol/L) group and TGF-β1 + CQ (60 μmol/L) group.Western blot was used to determine the expression of LC3-Ⅱ/LC3-Ⅰ,P62 and o-SMA in activated HSC-T6 cells.The expression of collagen Ⅰ and collagen Ⅲ was detected by immunocytochemical staining,Western blot and RT-qPCR.Western blot and RT-qPCR were also used to detect the expression of matrix metalloproteinase-13 (MMP-13),tissue inhibitor of metalloproteinase-1 (TIMP-1) and TIMP-2 at mRNA and protein levels.RESULTS:The ratio of LC3-Ⅱ / LC3-Ⅰ and P62 expression were increased after CQ intervention.Moreover,they were significantly higher in the TGF-β1 + CQ groups than those in TGF-β1 group (P ＜ 0.01).The expression of collagen Ⅰ and collagen Ⅲ at mRNA and protein levels was significantly increased in all TGF-β1 + CQ groups as compared with TGF-β1 group (P ＜0.01),and it was markedly increased among TGF-β1 + CQ groups in a dose-dependent manner.The expression of MMP-13 at mRNA and protein levels was significantly lowered and that of TIMP-1 and TIMP-2 was significantly increased in TGF-β1 + CQ groups as compared with TGF-β1 group (P ＜0.05).CONCLUSION:Inhibition of autophagy by CQ in activated HSC-T6 cells up-regulates the expression of collagen Ⅰ and collagen Ⅲ in a dose-dependent way,probably due to reduction of MMP-13 and enhancement of TIMP-1 and TIMP-2 expression.
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AIM:To explore the effects of chloroquine (CQ) on collagen Ⅰ and collagen Ⅲ expression in activated rat hepatic stellate cell line HSC-T6 and the possible mechanism.METHODS:Transforming growth factor-β1 (TGF-β1) was used to activate HSC-T6 cells and 3 doses of CQ was administered for 24 h.The cells were divided into 5 groups as follows:control group,TGF-β1 group,TGF-β1 + CQ (15 μmol/L) group,TGF-β1 + CQ (30 μ mol/L) group and TGF-β1 + CQ (60 μmol/L) group.Western blot was used to determine the expression of LC3-Ⅱ/LC3-Ⅰ,P62 and o-SMA in activated HSC-T6 cells.The expression of collagen Ⅰ and collagen Ⅲ was detected by immunocytochemical staining,Western blot and RT-qPCR.Western blot and RT-qPCR were also used to detect the expression of matrix metalloproteinase-13 (MMP-13),tissue inhibitor of metalloproteinase-1 (TIMP-1) and TIMP-2 at mRNA and protein levels.RESULTS:The ratio of LC3-Ⅱ / LC3-Ⅰ and P62 expression were increased after CQ intervention.Moreover,they were significantly higher in the TGF-β1 + CQ groups than those in TGF-β1 group (P ＜ 0.01).The expression of collagen Ⅰ and collagen Ⅲ at mRNA and protein levels was significantly increased in all TGF-β1 + CQ groups as compared with TGF-β1 group (P ＜0.01),and it was markedly increased among TGF-β1 + CQ groups in a dose-dependent manner.The expression of MMP-13 at mRNA and protein levels was significantly lowered and that of TIMP-1 and TIMP-2 was significantly increased in TGF-β1 + CQ groups as compared with TGF-β1 group (P ＜0.05).CONCLUSION:Inhibition of autophagy by CQ in activated HSC-T6 cells up-regulates the expression of collagen Ⅰ and collagen Ⅲ in a dose-dependent way,probably due to reduction of MMP-13 and enhancement of TIMP-1 and TIMP-2 expression.
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This study aimed to use the sika deer as a model to study the influence of IGF-1 on the expression of Col Ⅰ in antler chondrocytes.The chondrocytes were separated from sika deer antlers,cultured and were treated with recombinant human IGF-1 protein (rIGF-1),both rIGF-1 and PQ401,and transfected with IGF-1 over-expression plasmid or IGF-1 siRNA,respectively.The expression of Col Ⅰ which,a well-known marker for chondrocytes dedifferentiation,was detected by real-time PCR.The results showed that administration of rIGF-1 to antler chondroctyes resulted in an obvious decrease of Col Ⅰ mRNA levels,while PQ401 pretreatment could dramatically attenuate the effects of rIGF-1 on the expression of Col Ⅰ mRNA.After transfection with IGF-1 over-expression plasmid,the expression of Col Ⅰ mRNA was obviously reduced in antler chondrocytes compared with control.Conversely,knockdown IGF1 with specific siRNA could increase the expression of Col Ⅰ in antler chondrocytes.These results indicate that IGF-1 may play an important role in process of antler chondrocyte dedifferentiation.
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Objective To investigate the effect of estrogen on the fibrosis process of intrauterine adhesions and the expression of forkhead box F2 (FoxF2).Methods Primary human endometrial stromal cells (HESCs) were obtained by separation with 0.2％ collagenase Ⅰ digestion-mesh filtration-differential adherence,and identified by immunocytochemistry.HESCs affected with 10ng/ml transforming growth factor β1 (TGF-β1) for 48 hours.HESCs in model group were affected with 0,10-6,10-8,10-10 and 10-12mol/L estrogen,the expressions of smooth muscle actin alpha (α-SMA),Collagen I (COL Ⅰ) and FoxF2 were detected by quantitative PCR (qPCR) and Western blotting.Results HESCs with high purity and good activity were obtained by using 0.2％ collagenase Ⅰ digestion-mesh filtration-differential adherence separation method.Immunocytochemistry showed positive vimentin and negative cytokeratin 18 in HESCs.The results of qPCR and Western blotting showed that the mRNA and protein expression levels of α-SMA,COL Ⅰ and FoxF2 were higher in model group than in control group (P＜0.05),the model was built successfully.qPCR revealed that the mRNA expression levels ofα-SMA,COL Ⅰ and FoxF2 were significantly lower in 10-6,10-8 and 10-10mol/L estrogen groups than in model group (P＞0.05 in 10-10mol/L estrogen group,P＜0.05 in other groups),while in 10-12mol/L estradiol group,the expression levels of FoxF2 mRNA significantly decreased (P＜0.05),and of α-SMA and COL Ⅰ mRNA increased,but no significant difference were found (P＞0.05).Compared with the model group,the protein expression levels of α-SMA,COL Ⅰ and FoxF2 in 10-6,10-8 and 10 10mol/L estrogen groups decreased,but no significant difference was found (P＜0.05),while in 10-12mol/L estradiol group,the expression levels ofα-SMA protein increased (P＞0.05),and of COL Ⅰ and FoxF2 proteins decreased (P＜0.05).Conclusions The expression of FoxF2 in intrauterine adhesions is increased.Estrogen can reverse the fibrosis process of intrauterine adhesions in a certain range and inhibit the expression of FoxF2.
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Aim To investigate the effect of magnesi-um isoglycyrrhizinate (MgIG)on radiation -induced pulmonary fibrosis in mice and the mechanism.Meth-ods Fifty female C57BL/6 mice were randomly divid-ed into control group,irradiation (RT)group,MgIG group,RT +MgIG group and RT +dexamethasone (DXM)group,with 1 0 mice in each group.Except for control group and MgIG group,the remaining mice were given a single 1 5Gy 60 Co γray on whole lung. The mice in each group were administered 2 h before irradiation and each day after irradiation:MgIG group and RT +MgIG group were administered with MgIG (1 00 mg·kg -1 )by intraperitoneal injection;control group and RT group were administered with normal sa-line (20 mL·kg -1 )by intraperitoneal injection;RT+DXMgroup was administered with DXM(0.5 mg· kg -1 )by intraperitoneal injection.After 1 2 weeks,the mice were sacrificed and lung tissues were taken out. The degree of alveolitis and pulmonary fibrosis were observed by HE staining and Masson staining.The ex-pressions of type Ⅰ collagen,type Ⅲ collagen and TGF-β1 protein were detected by immunohistochem-isty.Results The alveolitis,pulmonary fibrosis and expressions of type Ⅰ collagen,type Ⅲ collagen, TGF-β1 ,p-Smad2,p-Smad3 increased significantly in RT group compared with control group (P <0.05 ), and were significantly lower in RT +MgIG group and RT +DXMgroup than those in RT group(P <0.05). Conclusion MgIG can improve radiation-induced pulmonary fibrosis in mouse lung tissue,and its mech-anism may be related to the influence of MgIG on TGF-βsignaling pathway.
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Background Sclera remodeling process in axial elongation is one of the main pathological mechanisms of axial myopia progression.Studies confirmed that transforming growth factor-β1(TGF-β1) participates in the sclera remodeling process,and Smad3 is one of TGF-β1 downstream signal gene transcriptive factors,so to explore its role in sclera remodeling process of myopic eyes is of great significance for pathogenesis and prevention research of myopia.Objective This study was to investigate the expressions of type Ⅰ collagen and Smad3,a TGF-31 downstream target,in sclera of form deprivation myopic (FDM) eyes and explore the impact of TGF-β1-Smad3-type Ⅰ collagen signaling pathway on collagen remodeling in myopic sclera.Methods Seventy-five 1-week-old guinea pigs were randomly divided into normal control group (25 guinea pigs) and FDM group (50 guinea pigs).Monocular FDM was induced by occluding the left eyes of guinea pigs in FDM group with translucent latex balloons for 2,4,6 weeks,respectively,and consecutive occluding for 4 weeks followed by uncovering for 1 week (4/-1 weeks).The refractive power was detected by retinoscopy and axial length was measured with A-type ultrasound.Immunohistochemistry and reverse transcription-PCR were employed to detect the dynamic expressions of type Ⅰ collagen and Smad3 protein ad mRNA in the sclera of guinea pigs with emmetropia and experimental myopia,ard the relationship between collagen Ⅰ and Smad3 levels was analyzed.Results The refraction was hypermetropic in both normal control group and FDM group before occluding of eyes (P＞0.05),and the hypermetropic power was gradually reduced over time in the normal control group.In the FDM group,the refractive power was gradually changed from (+2.09 ± 0.31)D before occluding to (-1.23±0.69),(-4.17±0.59),(-7.07±0.56) and (-4.30±0.58)D,and the axial length was increased from (5.93-±0.39)mm to (6.62±0.36),(7.30±0.34),(7.99--0.32),and (7.21 ±0.36) mm at weeks 2,4,6,and 4/-1 after occluding,respectively,indicating significant differences in refractive power and axial length over time in the FDM group from normal control group and self-control group (all at P＜0.05).The expressions of Smad3 and type Ⅰ collagen protein and mRNA in the sclera of the FDM group was significantly lower than those of the control group and self-control group in various time points (all at P＜0.05).The positive correlation were found in the expression of Smad3 on the myopic sclera with that of type Ⅰ collagen in both protein and mRNA levels (protein:r=0.993,P＜0.05;mRNA:r=0.954,P＜0.05).Conclusions The myopic power and ocular axis increase dependent upon occluding time,and the expressions of Smad3 and type Ⅰ collagen in the sclera are correspondingly weakened in FDM eyes.A consistent expression trend is found between Smad3 and type Ⅰ collage,suggesting Smad3 and type Ⅰ collagen participate in the regulation of sclera remodeling in myopia by TGF-β1-Smad3-Collagen Ⅰ signaling pathway.
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Objective:To investigate the effect of MT01 on the differentiation of osteoblasts under infected condition through determining the expression level of collagen Ⅰ (Col Ⅰ )mRNA in MG63 cells treated with Porphyromonas gingivalis (Pg).Methods:The cultured MG63 cells were divided into blank control,MT01,Pg, and MT01+Pg groups.MT01 at a concentration of 1 mg·L-1 was added into the MG63 cells,and the cells were incubated for 3 h.The cells treated with PBS (1 mg·L-1 )were used as control group.Then Pg (MOI=100∶1) was added.Real-time PCR was used to detect the expression levels of ColⅠ mRNA in MG63 cells at 2,4,6,8, 12 and 24 h after incubation.Results:Compared with blank control group,the levles of ColⅠ mRNA in the MG63 cells in MT01 and MT01 + Pg groups had no significant changes at 2 and 4 h (P > 0.05);the Col Ⅰ mRNA expression levels in MT01 group at 8,12 and 24 h were increased (P < 0.05 or P < 0.01).Compared with Pg group,the expression levels of ColⅠ mRNA in MT01+ Pg at 2 and 4 h were decreased,but the expression levels of ColⅠ mRNA were increased at 6,8,12 and 24 h (P <0.05 or P <0.01).Conclusion:MT01 can up-regulate the expression level of Col Ⅰ mRNA in the infected MG63 cells; MT01 could promot the differentiation of osteoblasts under infected condition.
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Objective To investigate the role of Rho kinase on collagen(COL)Ⅰ and Ⅲ expression of human renal tubular epithelial cells .Methods Human kidney tubular epithelial (HK‐2) cells were cultured in the RPMI‐1640 culture solution containing 15% fetal bovine serum .After 30 min pretreatment by the ALD receptor antagonist eplerenone(10 μmol/L) and Rho kinase inhibi‐tor Y27632(1 μmol/L) ,100 nmol/L ALD acted the HK‐2 cells for 24 h .The expressions of collagenⅠ and Ⅲ mRNA in each group were detected by real time PCR and the expression levels of COL Ⅰ ,Ⅲ and Rho kinase protein were detected by ELISA .Results ALD could up‐regulate the expressions of COLⅠ ,Ⅲ mRNA in HK‐2 cells ,and increased the levels of Rho kinase ,COLⅠ and Ⅲprotein ,while the Rho kinase inhibitor Y27632 and ALD receptor inhibitor eplerenone could antagonize these effects .Conclusion ALD could activate Rho kinase signal transduction pathway in HK‐2 cells and accelerate the progression of tubular interstitial fibro‐sis via Rho kinase induced expression of COL Ⅰ and Ⅲ .
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Objective To observe the prevention and treatment of the total flavonoids from Litchi chinensis Sonn( TFL) on hepatic fibrosis induced by dimethylnitrosamine(DMN)in rats, and to explore its mechanism. Methods Ninety SD rats were randomly divided into six groups, normal control group, model control group, colchicine group, high-, medium- and low-dose TFL group(n=15).Expect for normal control group, the other groups were given intraperitoneal injection of 2 mL.kg-1 of 5% dimethylnitrosamine for 4 weeks as the model group. The rats in the normal control group and model control group were given 5 mL.kg-1of 0.9% sodium chloride solution, colchicine group was treated with 0.1 mg.kg-1 colchicine.High-, medium-and low-dose TFL groups were given 200, 100 and 50 mg.kg-1 of TFL.The rats were sacrificed and the livers were harvested and stained with HE and Masson staining to observe pathological changes and liver fibrosis in the same part 6 weeks after all the medicine was given to the rats each day. Immunohistochemistry and Western blotting were used to detect the expression of the transforming growth factor β-Ⅰ/type Ⅱ receptor ( TβRⅠ/Ⅱ) , collagen Ⅰ( Col Ⅰ) and Ⅲ collagen ( Col Ⅲ) . Results Compared with the normal control group, the semiquantitative score of liver fiber and the protein expression of TβRⅠ, TβRⅡ, ColⅠ and Col Ⅲ in the model control group were significantly increased(P<0.01).Compared with the model control group, the protein expression levels of TβR, TβRⅡ, ColⅠand ColⅢwere significantly decreased( P<0.01) in the high-,medium-and low-dose TFL group.The semiquantitative score of liver fiber was significantly decreased( P<0.01) with a dose-effect relationship. Conclusion TFL can inhibit formation of DMN-induced liver fibrosis in rats, which may be related with reduction of expression of TβRⅠ/Ⅱ of hepatic fibrosis promoting factor TGF-β1 , inhibition of the activation and increase of hepatic stellate cells, reduction of the collagen content.