AIM： To investigate the effects of Danshao Huaxian （DSHX） capsules, a preparation of traditional Chinese medicine, on the expression of matrix metalloproteinase-1 （MMP-1）, and tissue inhibitor of metalloproteinas...AIM： To investigate the effects of Danshao Huaxian （DSHX） capsules, a preparation of traditional Chinese medicine, on the expression of matrix metalloproteinase-1 （MMP-1）, and tissue inhibitor of metalloproteinase-1 （TIMP-1） in the fibrous livers of rats. METHODS： Eighty male Wistar rats were randomly divided into normal control group （group A）, CCl4-induced hepatic fibrosis group （group B）, non-DSHX-treated group （group C）, low dose-treated group （group D）, and high dose-treated group （group E）. Fibrous liver models in rats were induced by subcutaneous injection of CCI4, oral administration of alcohol and high-lipid/low-protein diet for 8 wk. After the models were established, the rats in groups D and E were orally given a low dose （0.5 g/kg） and a high dose （1.0 g/kg） of DSHX daily for 8 wk, respectively. Then, the liver indexes, serum hyaluronic acid （HA） and alanine aminotransferase （ALT） were examined. The degree of hepatic fibrosis was evaluated by optical microscopy. Hydroxyproline （Hyp） in the urine was determined, and the expression of MMP-1 and TIMP-1 was detected by immunohistochemical techniques. RESULTS： In groups D and E, the liver indexes, levels of serum HA and ALT reduced and development of hepatic fibrosis weakened significantly. The urinary Hyp and expression of MMP-1 in the liver tissues elevated, but the expression of TIMP-1 decreased obviously, as compared to groups B and C.CONCLUSION： DSHX enhances the expression of MMP-1 but decreases that of TIMP-1 in liver tissues of CCI4-induced hepatic fibrotic rats, which may result in its elevated activity that contributes to fighting against hepatic fibrosis.展开更多
AIM:Matrix metalloproteinase-1 (MMP-1) and tissue inhibitor of matrix metalloproteinase-1 (TIMP-1) are central to the spontaneous resolution of liver fibrosis. The mechanisms involved have been investigated in hepatic...AIM:Matrix metalloproteinase-1 (MMP-1) and tissue inhibitor of matrix metalloproteinase-1 (TIMP-1) are central to the spontaneous resolution of liver fibrosis. The mechanisms involved have been investigated in hepatic stellate cells (HSC), but not in hepatocytes. We investigated the effects of fibril- and fixed-collagen on MMP-1 and TIMP-1 production in hepatocytes, using the HLE cell line. METHODS: Fibril type I and IV collagen were prepared by HCI digestion of type I and IV collagen, respectively. For fixed-collagen, culture dishes were coated with fibril type I or IV collagen and fixed by ultraviolet. Type I collagenase activity was measured using fluorescein isothiocyanate-labeled type I collagen. MMP-1 and TIMP-1 in HLE cells were measured by a one-step sandwich enzyme immunoassay. RESULTS: Both fibril type I and IV collagen significantly increased type I collagenase activity about two-fold compared with no fibril collagen. The effects of the fibril collagen were not affected by the coating condition. There was no significant difference in the effects on collagenase activity between cells cultured in medium containing fibril type I collagen and those cultured in the presence of type IV collagen. Both types of fibril collagen significantly increased MMP-1 production, and showed more than 10-fold higher levels of MMP-1 than the control. The enhanced MMP-1 production by fibril collagens was unaffected by the coating condition. By contrast, TIMP-1 production was not changed by the addition of fibril type I or IV collagen, and neither was it affected by the coating conditions. Coating with type I collagen significantly suppressed MMP-1 production by almost one-tenth compared with no coating. By contrast, TIMP-1 production was not affected by either the absence of a collagen coat or by increasing the concentration of the coating collagen. CONCLUSION: These results indicated that, in HLE cells, fibril- and fixed-collagen have opposite effects on MMP-1 production without affecting TIMP production. Fibril collagen in展开更多
[目的]探讨透明质酸钠(sod ium hyaluronate,SH)关节腔内注射治疗膝骨性关节炎(OA)的效果。[方法]24只兔子建立骨关节炎动物模型,随机分OA组、SH组、对照组,观察3组软骨、滑膜细胞病理切片及软骨MMP-1免疫组化,进行软骨M ank in s评分,...[目的]探讨透明质酸钠(sod ium hyaluronate,SH)关节腔内注射治疗膝骨性关节炎(OA)的效果。[方法]24只兔子建立骨关节炎动物模型,随机分OA组、SH组、对照组,观察3组软骨、滑膜细胞病理切片及软骨MMP-1免疫组化,进行软骨M ank in s评分,以及检测血液和关节液的IL-1含量的效果。[结果]SH组可见软骨破坏减轻,滑膜纤维增生减少,M ank in s评分有明显改善(P<0.05);血、膝关节滑液IL-1浓度降低(P<0.05),但关节软骨中MMP表达仍然活跃。[结论]SH能减少滑液中炎性介质IL-1的分泌,从而减轻滑膜炎症,缓解对软骨细胞的破坏,延缓OA进展,但仍阻止不了OA进展。展开更多
AIM: To investigate the expression and role of tissue inhibitor of matrix metalloproteinases-1 (TIMP-1) during natural aging in rat liver and to detect the expression of matrix metalloproteinase-2 (MMP-2) and MMP-9. M...AIM: To investigate the expression and role of tissue inhibitor of matrix metalloproteinases-1 (TIMP-1) during natural aging in rat liver and to detect the expression of matrix metalloproteinase-2 (MMP-2) and MMP-9. METHODS: The rats were divided into 3-mo-old group (n=5), 10-mo-old group (n=5) and 24-mo-old group (n=5). Histopathologic changes of liver were observed with HE and Masson stain. The location and protein expressions of TIMP-1 were determined by immunohistochemistry and Western blot; message RNA (mRNA) levels were measured in livers from rats of various ages by semi-quantitative reverse transcriptional polymerase chain reaction (RT PCR). In addition, the expression of MMP-2 and MMP-9 was assessed by RT-PCR and Western blot. RESULTS: Histologic examination showed that the aging liver had excessive fatty degeneration and collagen deposition. Immunohistochemical staining showed that TIMP-1 related antigen in livers was located in cytoplasm. The protein expression of TTMP-1 was significantly higher in the oldest animals and the mRNA expression was increased significantly in the 24-mo-old rats (t= 4.61, P= 0.002<0.05, 24-vs 10-mo-old rats; t=4.31, P=0.003<0.05, 24- vs 3-mo-old rats). The expression of MMP-2 and MMP-9 had no change during aging; the ratios TIMP-1/MMP-2 and TIMP-1/MMP-9 in aging liver were significantly higher than those in maturation and young livers. CONCLUSION: TIMP-1 may play an important role in the process of liver aging.展开更多
Objective: To observe the influence of intra-articular injection of sodium hyaluronate (HA) on the mRNA expressions of matrix metalloproteinase-1,-3 (MMP-1,-3) and tissue inhibitor of metalloproteinase-1 (TIMP-1) in c...Objective: To observe the influence of intra-articular injection of sodium hyaluronate (HA) on the mRNA expressions of matrix metalloproteinase-1,-3 (MMP-1,-3) and tissue inhibitor of metalloproteinase-1 (TIMP-1) in cartilage and synovium of traumatic osteoarthritis (OA).Methods: Sixteen white rabbits underwent unilateral anterior cruciate ligament transection (ACLT) were divided into 2 groups randomly 5 weeks after transection. The experimental group rabbits received 0.3 ml of 1% HA by intra-articular injection once a week. Animals in the control group were treated under the same conditions using physiological saline. Ten weeks following surgery, cartilage and synovium were harvested. The mRNA expressions of MMP-1, MMP-3 and TIMP-1 were analyzed using reverse transcription-polymerase chain reaction (RT-PCR).Results: In synovium, the mRNA expression of MMP-3 was suppressed in the HA injection group. HA treatment had no effect on the MMP-3 expression in cartilage. No significant difference of MMP-1 and TIMP-1 expressions in cartilage and synovium was found between the HA injection group and the control group.Conclusions: One of the mechanisms of the therapeutic effect of HA may be the inhibition of expression of MMP-3 in synovium during early stage of traumatic OA.展开更多
基金Supported by the Foundation of Traditional Chinese Medicine Modernization Project of Guizhou Province, No. 200409
文摘AIM： To investigate the effects of Danshao Huaxian （DSHX） capsules, a preparation of traditional Chinese medicine, on the expression of matrix metalloproteinase-1 （MMP-1）, and tissue inhibitor of metalloproteinase-1 （TIMP-1） in the fibrous livers of rats. METHODS： Eighty male Wistar rats were randomly divided into normal control group （group A）, CCl4-induced hepatic fibrosis group （group B）, non-DSHX-treated group （group C）, low dose-treated group （group D）, and high dose-treated group （group E）. Fibrous liver models in rats were induced by subcutaneous injection of CCI4, oral administration of alcohol and high-lipid/low-protein diet for 8 wk. After the models were established, the rats in groups D and E were orally given a low dose （0.5 g/kg） and a high dose （1.0 g/kg） of DSHX daily for 8 wk, respectively. Then, the liver indexes, serum hyaluronic acid （HA） and alanine aminotransferase （ALT） were examined. The degree of hepatic fibrosis was evaluated by optical microscopy. Hydroxyproline （Hyp） in the urine was determined, and the expression of MMP-1 and TIMP-1 was detected by immunohistochemical techniques. RESULTS： In groups D and E, the liver indexes, levels of serum HA and ALT reduced and development of hepatic fibrosis weakened significantly. The urinary Hyp and expression of MMP-1 in the liver tissues elevated, but the expression of TIMP-1 decreased obviously, as compared to groups B and C.CONCLUSION： DSHX enhances the expression of MMP-1 but decreases that of TIMP-1 in liver tissues of CCI4-induced hepatic fibrotic rats, which may result in its elevated activity that contributes to fighting against hepatic fibrosis.
文摘AIM:Matrix metalloproteinase-1 (MMP-1) and tissue inhibitor of matrix metalloproteinase-1 (TIMP-1) are central to the spontaneous resolution of liver fibrosis. The mechanisms involved have been investigated in hepatic stellate cells (HSC), but not in hepatocytes. We investigated the effects of fibril- and fixed-collagen on MMP-1 and TIMP-1 production in hepatocytes, using the HLE cell line. METHODS: Fibril type I and IV collagen were prepared by HCI digestion of type I and IV collagen, respectively. For fixed-collagen, culture dishes were coated with fibril type I or IV collagen and fixed by ultraviolet. Type I collagenase activity was measured using fluorescein isothiocyanate-labeled type I collagen. MMP-1 and TIMP-1 in HLE cells were measured by a one-step sandwich enzyme immunoassay. RESULTS: Both fibril type I and IV collagen significantly increased type I collagenase activity about two-fold compared with no fibril collagen. The effects of the fibril collagen were not affected by the coating condition. There was no significant difference in the effects on collagenase activity between cells cultured in medium containing fibril type I collagen and those cultured in the presence of type IV collagen. Both types of fibril collagen significantly increased MMP-1 production, and showed more than 10-fold higher levels of MMP-1 than the control. The enhanced MMP-1 production by fibril collagens was unaffected by the coating condition. By contrast, TIMP-1 production was not changed by the addition of fibril type I or IV collagen, and neither was it affected by the coating conditions. Coating with type I collagen significantly suppressed MMP-1 production by almost one-tenth compared with no coating. By contrast, TIMP-1 production was not affected by either the absence of a collagen coat or by increasing the concentration of the coating collagen. CONCLUSION: These results indicated that, in HLE cells, fibril- and fixed-collagen have opposite effects on MMP-1 production without affecting TIMP production. Fibril collagen in
文摘[目的]评价5氟脲嘧啶(5-F luorourac il,5Fu)关节腔注射治疗兔膝骨性关节炎(osteoarthritis,OA)的疗效。[方法]24只兔子制成骨关节炎模型随机分成OA组、5Fu组和对照组,OA组立即处死,5Fu组按5Fu 2mg/kg关节腔注射,每周1次连续4次,对照组注射等量生理盐水,最后1次治疗后1周处死。观察3组滑膜组织的光镜、电镜改变及软骨的光镜、MMP-1免疫组化改变,比较软骨M ank in s评分及关节液中IL-1的浓度。[结果]5Fu组可见软骨破坏减轻,滑膜炎症明显抑制,M ank in s评分明显改善(P<0.01);关节液IL-1浓度降低(P<0.05),关节软骨中MMP-1表达减弱。[结论]5Fu关节腔内注射能抑制滑膜炎症,缓解软骨的破坏。
文摘[目的]探讨透明质酸钠(sod ium hyaluronate,SH)关节腔内注射治疗膝骨性关节炎(OA)的效果。[方法]24只兔子建立骨关节炎动物模型,随机分OA组、SH组、对照组,观察3组软骨、滑膜细胞病理切片及软骨MMP-1免疫组化,进行软骨M ank in s评分,以及检测血液和关节液的IL-1含量的效果。[结果]SH组可见软骨破坏减轻,滑膜纤维增生减少,M ank in s评分有明显改善(P<0.05);血、膝关节滑液IL-1浓度降低(P<0.05),但关节软骨中MMP表达仍然活跃。[结论]SH能减少滑液中炎性介质IL-1的分泌,从而减轻滑膜炎症,缓解对软骨细胞的破坏,延缓OA进展,但仍阻止不了OA进展。
基金Supported by the Major State Basic Research and Development Program of China, 973 Program, No.G2000057000 Creative Research Group Fund of the National Science Foundation of China, No.30121005the Postdoctor Foundation of China, No. 2004035048
文摘AIM: To investigate the expression and role of tissue inhibitor of matrix metalloproteinases-1 (TIMP-1) during natural aging in rat liver and to detect the expression of matrix metalloproteinase-2 (MMP-2) and MMP-9. METHODS: The rats were divided into 3-mo-old group (n=5), 10-mo-old group (n=5) and 24-mo-old group (n=5). Histopathologic changes of liver were observed with HE and Masson stain. The location and protein expressions of TIMP-1 were determined by immunohistochemistry and Western blot; message RNA (mRNA) levels were measured in livers from rats of various ages by semi-quantitative reverse transcriptional polymerase chain reaction (RT PCR). In addition, the expression of MMP-2 and MMP-9 was assessed by RT-PCR and Western blot. RESULTS: Histologic examination showed that the aging liver had excessive fatty degeneration and collagen deposition. Immunohistochemical staining showed that TIMP-1 related antigen in livers was located in cytoplasm. The protein expression of TTMP-1 was significantly higher in the oldest animals and the mRNA expression was increased significantly in the 24-mo-old rats (t= 4.61, P= 0.002<0.05, 24-vs 10-mo-old rats; t=4.31, P=0.003<0.05, 24- vs 3-mo-old rats). The expression of MMP-2 and MMP-9 had no change during aging; the ratios TIMP-1/MMP-2 and TIMP-1/MMP-9 in aging liver were significantly higher than those in maturation and young livers. CONCLUSION: TIMP-1 may play an important role in the process of liver aging.
文摘Objective: To observe the influence of intra-articular injection of sodium hyaluronate (HA) on the mRNA expressions of matrix metalloproteinase-1,-3 (MMP-1,-3) and tissue inhibitor of metalloproteinase-1 (TIMP-1) in cartilage and synovium of traumatic osteoarthritis (OA).Methods: Sixteen white rabbits underwent unilateral anterior cruciate ligament transection (ACLT) were divided into 2 groups randomly 5 weeks after transection. The experimental group rabbits received 0.3 ml of 1% HA by intra-articular injection once a week. Animals in the control group were treated under the same conditions using physiological saline. Ten weeks following surgery, cartilage and synovium were harvested. The mRNA expressions of MMP-1, MMP-3 and TIMP-1 were analyzed using reverse transcription-polymerase chain reaction (RT-PCR).Results: In synovium, the mRNA expression of MMP-3 was suppressed in the HA injection group. HA treatment had no effect on the MMP-3 expression in cartilage. No significant difference of MMP-1 and TIMP-1 expressions in cartilage and synovium was found between the HA injection group and the control group.Conclusions: One of the mechanisms of the therapeutic effect of HA may be the inhibition of expression of MMP-3 in synovium during early stage of traumatic OA.
