ABSTRACT
To investigate effectsof Yangyinyiqi Mixture on pulmonary fibrosis caused by bleomycin. SD ratswere divided randomly into: model group(distilled water,1 mL·0.1 kg-1), dexamethasone acetate group (dexamethasone acetate, the dosage was reduced gradually), low-dose group (Yangyinyiqi Mixture, 11 g·kg-1), moderate-dose group (Yangyinyiqi Mixture, 22 g·kg-1), high-dose group (Yangyinyiqi Mixture, 44 g·kg-1) and control group (distilled water, 1 mL·0.1 kg-1). Yangyinyiqi Mixture and dexamethasone acetate were intragastrically administrated. Lung tissue was collected for histopathological examination. Compared with control group, collagen markedly increased and HYP content significantly increased on 7th day in model group (p<0.01). On 28th day, collagen was diffusely deposited, alveolar was destroyed, and HYP content significantly increased (p<0.01). Compared with model group, bleomycin-induced suffering injury caused MMP-9 expression levels to rapidly increase (7and 14 days, p<0.01). TIMP-1 markedly increased (7and 14 days, p<0.01) and stayed at a high level to28th day. Yangyinyiqi Mixture exerted an effect against pulmonary fibrosis, which could involved prevention of collagen deposition through inhibitingMMP-9 and TIMP-1 expression.
El trabajo investiga los efectos de la mezcla Yangyinyiqi sobre la fibrosis pulmonary causada por bleomicina. Ratas SD se dividieron aleatoriamente en: grupo modelo (agua destilada, 1 mL·0.1 kg-1), grupo acetate de dexametasona (acetate de dexametasona, la dosis se redujo gradualmente), grupo de dosis baja (mezcla Yangyinyiqi, 11 g·kg-1), grupo de dosis moderada (mezcla Yangyinyiqi, 22 g·kg-1), grupo de dosis alta (mezcla Yangyinyiqi, 44 g·kg-1) y grupo control (agua destilada, 1 Ml·0.1 kg-1). La mezcla de Yangyinyiqi y el acetate de dexametasona se administraron por vía intragástrica. Se recolectó tejido pulmonary para examen histopatológico. En comparación con el grupo control, el colágeno aumentó notablemente y el contenido de HYP aumentó significativamente el séptimo día en el grupo modelo (p<0.01). El día 28, el colágeno se depositó difusamente, se produjo destrucción alveolar y el contenido de HYP aumento significativamente (p<0.01). En comparación con el grupo modelo, la lesión inducida por bleomicina causó que los niveles de expression de MMP-9 aumentaron rápidamente (7 y 14 días, p<0.01). TIMP-1 aumentó notablemente (7 y 14 días, p<0.01) y se mantuvo en un nivel alto hasta el día 28. La mezcla Yangyinyiqi ejerció un efecto contra la fibrosis pulmonary, lo que podría implicar la prevención del deposito de colágenio mediante la inhibición de la expression de MMP-9 y TIMP-1.
Subject(s)
Animals , Male , Rats , Pulmonary Fibrosis/drug therapy , Drugs, Chinese Herbal/administration & dosage , Tissue Inhibitor of Metalloproteinases/metabolism , Matrix Metalloproteinase 9/metabolism , Bleomycin , Dexamethasone/administration & dosage , Blotting, Western , Rats, Sprague-Dawley , Matrix Metalloproteinase 1 , Disease Models, Animal , Hydroxyproline/analysisABSTRACT
BackgroundObstruction of aqueous humor out flow pathway or abnormality of the extracellular matrix( ECM ) of trabecular meshwork cells causes high intraocular pressure. The balance of matrix metalloproteinases (MMPs) and tissue inhibitors of matrix metalloproteinases(TIMPs) is critical for the metabolism of ECM. Interleukin1α(IL-1α) can influence outflow of aqueous humor by regulating MMPs level. Objective This study was to investigate the effect of interleukin-1α on the expression of MMP-2,MMP-3 and TIMP-I in cultured swine trabecular meshwork cells.Methods Swine sclera with trabecular meshwork tissue was isolated from 20 swine eyes and cultured with explant cultured method. Cultured cells were passaged and third generation cells were identified by fibronectin ( FN ) and laminin ( LN ) staining. After 24 hours of serum starvation, trabecular meshwork cells treated with IL-1α at the concentration of 10 mg/L were regarded as the IL group,and serum-free culture medium used to treat trabecular meshwork cells was regarded as the control group. The expression of MMP-2, MMP-3 and TIMP-1 proteins in trabecular meshwork cells were detected by immunohistochemistry,and the expression of MMP-2 mRNA, MMP-3 mRNA and TIMP-1 mRNA were detected by RT-PCR. The examination results were compared between the two groups. ResultsThe third generation of cells were positive for FN and LM. Compared with the control group, the expression levels of MMP-3 and TIMP-1 proteins(A value) in trabecular meshwork cells were significantly higher in the IL group than the control group(t=-7. 694,t =-5. 199,P＜0. 05) ,but no obvious difference was found in the expression of MMP-2 between the two groups( t=-2. 365, P＞0.05 ). The higher expression levels in MMP-3 mRNA and TIMP-1 mRNA (A value) in trabecular meshwork cells were seen in comparison with the control group (t =-3. 025,t=-1. 921 ,P＜0. 05). However,similar results were found in the expression of MMP-2 mRNA between the two groups(t =- 1. 173, P＞0.05 ). ConclusionsThe overexpression of MMP-3 and TIMP-1 proteins and their mRNA leads to the imbalance of MMP-3/TIMP-1 and promotes the decomposition of ECM in the trabecular meshwork, and therefore increases aqueous outflow.
ABSTRACT
Objective To investigate the expression of MMP-2, MMP-9 and TIMP-1. TIMP-2 during the course of traumatie pro- liferative vitreoretinopathy (tPVR) in rat retina treated with GM6001 and without GM6001 and evaluate the interventiunal effect uf GM6001 on tPVR. Design Experimental study. Participants 108 SD rats. Methods Rats were divided randomly into three groups: the Ns control group, the tPVR group, the tPVR treated with GM6001 group. The expression of MMP-2. MMP-9 and TIMP-1. TIMP-2 in retina was analyzed by Western Blot on day 1, 3. 7, 14, 21 and 28. Main Outcome Measures The expression of MMP-2, MMP-9, TIMP-2, TIMP-1 in the SD rats' retina of every group. Results The results of Western Blot showed that the expression of MMP-2 in the SD rats' retina of the tPVR group was stronger than the one in other groups at day 3, 7, 14, 21 and 28: the tPVR treated with GM6001 group was weaker than the one in the tPVR group at day 14, 21 and 28d. The expression of MMP-9 in the tPVR group was stronger than the one in other groups at all time; the cireumstance of tPVR treated with GM6001 group was weaker than the one in the tPVR group at day 1, 3, 7, 14 and 21. The rate of MMP-2/TIMP-2 of the SD rats" retina in the tPVR group increased at day 14, 21 and 28, and it was lower in the tPVR treated with GM6001 group compared with the tPVR group. The rate of MMP-9/TIMP-1 in the tPVR group increased at all time, and it degraded in the tPVR treated with GM6001 group as compared with the tPVR group. Conclusions The dishalance of MMP-2 and TIMP-2, MMP-9 and TIMP-1 involves with the course of tPVR. GM6001 playes an important role in in- terfering the course of tPVR by regulating the balance of these cell faetors.
ABSTRACT
The enhanced process of proteolysis of both the basement membrane and the stromal extracelluar matrix (ECM) contributes to the escape of breast cancer cells into the neighboring tissues, eventually leading to the formation of distant metastases. A group of enzymes thought to play a role in tumor cell invasion are the matrix metalloproteinases (MMPs). Much attention has been focused on MMP-2 and MMP-9, which are 2 members of the MMP family active against collagen of the basement membrane. The enzymatic activities of MMP-2 and MMP-9 are inhibited by the tissue inhibitors of metalloproteinases (TIMPs). TIMP-2, one member of TIMPs, inhibits MMP-2 and MMP-9. The imbalance between TIMPs and MMPs permits to tumor invasion and metastasis. Theretore, TIMPs constitute promising targets in the developmemt of anticancer terapies. Immunohistological stainings of MMP-2, MMP-9 and TIMP-2 were performed on paraffin-embedded tissue sections of 31 invasive breast carcinomas. MMP-2 and MMP-9 were associated with neoplastic cell cytoplasms in 65% of the cases and exhibited inter-tumoral variability of the staining intensity. The MMP-2 and MMP-9 stainings did not correlate with presence of metastases at time of diagnosis. TIMP-2 was detected in the peri-tumoral stroma and was present in 81% of the cases. Residual benign breast tissue was negative for TIMP-2 staining. Neoplasms with diffuse TIMP-2 staining (32%) have metastasis significantly more frequently (50% metastasis) than ceses with focal (20% metastasis) or absent (0% metastasis) TIMP-2. We conclude that the clinical outcome such as metastasis is more closely related to the presence of TIMP-2 than the corresponding MMPs. Enhanced TIMP-2 expression, therefore, may denote a stromal response to tumor invasion, indicative of aggressive behavior in the subset breast carcinoma.