Decreased Expression of Canstatin in Rat Model of Monocrotaline-Induced Pulmonary Arterial Hypertension: Protective Effect of Canstatin on Right Ventricular Remodeling.

Pulmonary arterial hypertension (PAH) is a progressive disease which causes right ventricular (RV) failure. Canstatin, a C-terminal fragment of type IV collagen α2 chain, is expressed in various rat organs. However, the expression level of canstatin in plasma and organs during PAH is still unclear. We aimed to clarify it and further investigated the protective effects of canstatin in a rat model of monocrotaline-induced PAH. Cardiac functions were assessed by echocardiography. Expression levels of canstatin in plasma and organs were evaluated by enzyme-linked immunosorbent assay and Western blotting, respectively. PAH was evaluated by catheterization. RV remodeling was evaluated by histological analyses. Real-time polymerase chain reaction was performed to evaluate RV remodeling-related genes. The plasma concentration of canstatin in PAH rats was decreased, which was correlated with a reduction in acceleration time/ejection time ratio and an increase in RV weight/body weight ratio. The protein expression of canstatin in RV, lung and kidney was decreased in PAH rats. While recombinant canstatin had no effect on PAH, it significantly improved RV remodeling, including hypertrophy and fibrosis, and prevented the increase in RV remodeling-related genes. We demonstrated that plasma canstatin is decreased in PAH rats and that administration of canstatin exerts cardioprotective effects.

Effects of herbal compound 861 on collagen synthesis and degradation in rat mesangial cells exposed to high glucose.

OBJECTIVE: To investigate the effects of Herbal Compound 861 (Cpd 861) on collagen synthesis and degradation in rat mesangial cells exposed to high glucose. METHODS: The third to fifth passage of rat mesangial cells were exposed to high glucose and Cpd 861 at a concentration of 0.25-4.00 g/L for 24, 48 and 72 h, respectively. Benazepril (10(-7)-10(-3) mmol/L) was selected as positive control. The methyl thiazolyl tetrazolium colorimetric assay was used to evaluate the effect of Cpd 861 on cell proliferation. After incubation with Cpd 861 at a concentration of 2.00 g/L for 48 h, the protein secretions of collagen type IV, matrix metallopeptidase 9 (MMP-9), tissue inhibitor of metalloproteinase-1 (TIMP-1), transforming growth factor beta 1 (TGF-ß1), and hepatocyte growth factor (HGF) were detected by enzyme-linked immunosorbent assay method. And rat mesangial cells were harvested to determine MMP-9, TIMP-1, TGF-ß1 and HGF mRNA expression by reverse transcription polymerase chain reaction. RESULTS: Cpd 861 inhibited cell proliferation induced by high glucose in a dose- and time-dependent manner. Compared with high glucose, collagen type IV production was decreased significantly by Cpd 861 (P<0.01). Cpd 861 increased the protein secretions and mRNA expressions of MMP-9 and HGF, whereas the protein secretions and mRNA expressions of TIMP-1 and TGF-ß1 were reduced markedly (P<0.05). The ratio of MMP-9 to TIMP-1 was enhanced by Cpd 861 significantly. There was no significant difference in all above-mentioned effects between Cpd 861 (2.00 g/L) and benazepril (10(-5) mmol/L). CONCLUSION: The anti-glomerulosclerosis mechanisms of Cpd 861 were partly attributed to its effects of inhibiting mesangial cell proliferation, decreasing collagen synthesis and enhancing collagen degradation.

Effect of topical application of chlorogenic acid on excision wound healing in rats.

This study was undertaken to evaluate the therapeutic effects of topical chlorogenic acid on excision wounds in Wistar rats. A 1 % (w/w) chlorogenic acid or silver sulfadiazine ointment was applied topically once a day for 15 days on full-thickness excision wounds created on rats. The 1 % (w/w) chlorogenic acid ointment had potent wound healing capacity as evident from the wound contraction on the 15th post-surgery day, which was similar to that produced by 1 % (w/w) silver sulfadiazine ointment. Increased rates of epithelialization were observed in the treated rats. It also improved cellular proliferation, increased tumor necrosis factor-α levels during the inflammatory phase (12 h, 24 h, 48 h, and 72 h post-wounding) of wound healing, upregulated transforming growth factor-ß1 and elevated collagen IV synthesis in the chlorogenic acid-treated group. The results also indicated that chlorogenic acid possesses potent antioxidant activity by increasing superoxide dismutase, catalase, and glutathione, and decreasing lipid peroxidation. In conclusion, these results demonstrate that topical application of chlorogenic acid can accelerate the process of excision wound healing by its ability to increase collagen synthesis through upregulation of key players such as tumor necrosis factor-α and transforming growth factor-ß1 in different phases of wound healing as well as by its antioxidant potential.

[Effect of compound Puerarin on the collage IV in streptozotocin-induced diabetic nephropathy rats].

OBJECTIVE: To observe the effect of compound Puerarin on collagen IV of streptozotocin-induced diabetic rats. METHODS: Diabetic nephropathy rats were induced by intraperitoneal injection of streptozotocin (STZ). Rats were allocated randomly to control group (10), diabetes model group (10), Vitamin C group (10), Puerarin group (10), vitamin C plus Puerarin group (10). The study period lasted for 12 weeks. During and after the treatment, the general state, blood glucose levels, glycosylated hemoglobin, blood urea nitrogen, serum collagen IV, blood urea nitrogen, serum creatinine, urinary albumin excretion rate of the 24-hour, and clearance rate of creatinine collagen IV protein were determined by immunohistochemistoche analysis as well as type the gene expression of collagen IV alpha 1 mRNA were determined by in situ hybridization analysis in the kidney tissue of different groups. RESULTS: (1) Diabetes mellitus and renal function lesion occurred in the four groups. (2) Vitamin C and Puerarin could improve the general conditions of diabetic Rats, decrease blood urea nitrogen [(8.68 +/- 0.43), (7.98 +/- 0.47) and (5.76 +/- 0.82) micromol/L, serum creatinine [(74.68 +/- 8.20), (75.52 +/- 7.98) and (58.66 +/- 6.65) mmol/L], and urinary albumin excretion rate of the 24-hour [(18.40 +/- 0.37), (17.24 +/- 0.30) and (9.97 +/- 1.27) mg/24 h x 10(-3)]; increase clearance rate of creatinine [(0.59 +/- 0.21), (0.61 +/- 0.14) and (0.69 +/- 0.32) ml/min], the expression of collage IV absorbance [(111.56 +/- 14.61), (110.78 +/- 9.69) and (95.44 +/- 9.97) ] in the diabetic Rats were significantly inhibited at the same time. CONCLUSION: The compound Puerarin might have some functions on preventing ren by inhibiting expression of type IV collagen.

[Mechanism of the effect of Tongluo Recipe against glomerular sclerosis in rats].

OBJECTIVE: To investigate the effects of Tongluo Recipe on the expression of collagen IV (Col IV), fibronectin (FN), laminin (LN), transforming growth factor-beta1 (TGF-beta1) in rat renal tissues and explore the mechanism underlying these effects in rats with glomerular sclerosis. METHODS: The pathological changes in the renal tissues of rats with glomerular sclerosis were observed microscopically, and the expressions of Col IV, FN, LN, and TGF-beta1 were detected using immunohistochemical staining and image analysis system. RESULTS: Tongluo Recipe significantly decreased the expressions of Col IV, FN, LN and TGF-beta1 in the renal tissue of rats with glomerular sclerosis (P<0.05 or P<0.01) and obviously alleviated the renal pathologies (P<0.01). CONCLUSION: The therapeutic effects of Tongluo Recipe are probably mediated by lowered expressions of Col IV, FN, LN and TGF-beta1.

Regression of glomerulosclerosis in subtotally nephrectomized rats: effects of monotherapy with losartan, spironolactone, and their combination.

Angiotensin II accelerates and renin-angiotensin system blockade halts progression; blockade with high doses even reverses established glomerulosclerosis. Aldosterone also accelerates progression of glomerulosclerosis, partially independently of angiotensin II. The purpose of this study was to assess the relative ability of an angiotensin receptor type 1 (AT1) blocker, a mineralocorticoid receptor blocker, and their combination to reverse glomerulosclerosis. Sprague-Dawley rats were subjected to subtotal renal ablation (SNX) or sham operation. Eight weeks after surgery, they were either euthanized or allocated to treatment with vehicle, losartan, spironolactone, their combination, or unspecific antihypertensive treatment (dihydralazine) for 4 wk. Renal morphology was evaluated by stereology in tissues obtained using pressure-controlled perfusion fixation. Systolic blood pressure was significantly higher in SNX compared with sham-operated animals and decreased in all treatment groups. Compared with wk 8 after SNX, the glomerulosclerosis index (GSI) had increased further by week 12 in the vehicle- and dihydralazine-treated groups but was significantly lowered in the SNX+losartan as well as in the SNX+losartan+spironolactone groups and had not progressed further in the SNX+spironolactone group. The study confirms the partial regression of established glomerulosclerosis in subtotally nephrectomized rats after high-dose AT1 receptor blockade. Nonhyperkalemic doses of spironolactone prevented the increase but failed to decrease the GSI below the 8-wk level and preserved podocyte numbers. Combining the AT1 blocker with mineralocorticoid receptor blockade failed to further increase the regression of glomerulosclerosis.

[Effects of Cordyceps sinensis on dimethylnitrosamine-induced liver fibrosis in rats].

OBJECTIVE: To study the effects of Cordyceps sinensis on dimethylnitrosamine-induced liver fibrosis in rats. METHODS: SD rats were divided into normal control group, untreated group and Cordyceps sinensis-treated group. The rats in each group were fed with corresponding drug for 4 weeks. The rat's liver collagen deposition was observed with collagen staining. Hydroxyproline (Hyp) contents in liver tissue of the rats in 3 groups were determined with HCl hydrolysis. The tissue inhibitor of metalloproteinase-2 (TIMP-2) and type IV collagen contents were observed by Envision, and matrix metalloproteinases-2 (MMP-2) activity was detected by the method of enzyme-picture. Type I collagen was detected by Western blotting. RESULTS: The contents of Hyp, TIMP-2, type IV collagen, and the expression of type I collagen in untreated group were significantly higher than those in the normal control group, while those in Cordyceps sinensis-treated group were significantly lower than those in the untreated group. The content of MMP-2 in untreated group was significantly lower than that in the normal control group, while that in Cordyceps sinensis-treated group was significantly higher than that in the untreated group. CONCLUSION: Cordyceps sinensis can considerably relieve the liver fibrosis, and the mechanism may be related to promoting the degradation of the collagens.

[Effect of anti-fibrosis compound contained serum on procollagen Type I and IV, matrix metalloproteinase and its tissue inhibitor-1 gene expression in HSC-LI90 cell line].

OBJECTIVE: To study the effects of Anti-fibrosis Compound contained serum (AFCS) on procollagen type I and IV (ProC-I and ProC-IV), matrix metalloproteinase (MMP) and its tissue inhibitor (TIMP-1) gene expression in hepatic stellate cell line LI90 (HSC-LI90). METHODS: AFCS was prepared by gastric infusing different dosage (0.5 g/kg, 2.0 g/kg and 4.0 g/kg) of Anti-fibrosis Compound Recipe to rats. After HSC-LI90 cells were exposed to AFCS for 48 hrs, levels of ProC-I, ProC-IV, gene expression of MMP-2, MMP membrane type 1 (MT1-MMP) and TIMP-1 in the cells were detected by Northern blot, and gelatinase activity of MMP-2 was measured by zymography. RESULTS: AFCS of different concentrations could inhibit ProC-I and ProC-IV and TIMP-1 gene expression (P < 0.05 or P < 0.01), increase MT1-MMP gene expression (P < 0.01), but it showed no effect on gene expression and activity of MMP-2 (P > 0.05). CONCLUSION: Anti-fibrosis Compound Recipe has anti-liver fibrosis action, its effects in inhibiting TIMP-1 gene expression of HSC-LI90 cells and promoting degradation of collagen might be one of the mechanisms of the action.

[Experimental study of compound salvia injection in preventing and treating chronic nephrotoxicity induced by cyclosporin A in rats].

OBJECTIVE: To explore the mechanism of protective effect of Compound Salvia Injection (CSI) on experimental cyclosporin A induced nephrotoxicity. METHODS: Rats were on low-salt diet and cyclosporin A (CsA) was administered once a day through gastrogavage at dosage of 30 mg/kg.d for 28 days. Expression of the mRNA for intrarenal transforming growth factor-beta 1 (TGF-beta 1) and renin was measured by reverse transcription-polymerase chain reaction (RT-PCR). Intrarenal expression of TGF-beta 1 and Collagen IV was determined by immunohistochemical assays. The effects of CSI on these changes were also evaluated. RESULTS: Chronic CsA-induced nephropathy might be correlated to TGF-beta 1 and renin mRNA up-regulation as well as matric proteins accumulation in interstitium. CSI could reduce these changes. CONCLUSION: Decreased CsA-related TGF-beta 1 and renin upregulation expression and accumulation of matrix proteins in the kidney might be related to the protective mechanism of CSI on CsA-induced chronic nephrotoxicity.

[Effects of cyclosporin A combined with T4 on collagen IV production, coll alpha 1 (IV) mRNA and TGF beta 1 mRNA expression in cultured human mesangial cells].

OBJECTIVE: To investigate the effects of single cyclosporin A (CsA), tripcholorlide (T4) or both on renal fibrosis. METHODS: Collagen IV concentrations in the supernatants of human mesangial cells (HMCs) in culture were measured by ELISA. Collagen type IV alpha 1 mRNA (Coll alpha 1 IV mRNA) and transforming growth factor beta 1 (TGF beta 1) mRNA expression were tested by RT-PCR. RESULTS: CsA regulated HMC collagen IV production biphysically, i.e., 1.0 microgram/ml CsA increased collagen synthesis while 0.01-0.1 microgram/ml CsA decreased its formation. T4 enhanced collagen IV production in a dose dependent manner. Combination of the two drugs in lower doses could reach the same degree of immunosuppression as a higher dose of CsA did but with less fibrogenesis. Both CsA and T4 up-regulated HMC Coll alpha 1 (IV) mRNA and TGF beta 1 mRNA expression. The degree of Coll alpha 1 (IV) mRNA expression in different groups were in accordance with that of TGF beta 1 mRNA. CONCLUSIONS: Both CsA and T4 regulate extracellular matrix formation which may be mediated by TGF beta 1. Combination of the two drugs in lower dose results in less fibrosis than a higher dose of CsA.