ABSTRACT
Background Anti-scarring is a focus following glaucoma filtering surgery.It has been verified that expression of heat shock protein 47 (HSP47) is always correlated with that of various types of collagens and closely related with the collagen-related diseases including fibrosis in various organs.However,the relationship between the expression of HSP47 and fibrosis of filtering bleb is unclear.Ohjective Present study was to investigate the dynamic expression of HSP47 in subconjunctival filtering bleb after anti-glaucomatous filtering surgery.Methods Filtering surgery was performed on the right eyes of 10 SPF male Spargue-Dawley(SD) rats.Then the operative eyes were randomized into the postoperative 7-day group and 14-day group according to the randomized number table,and the left eyes of the rats served as the normal control group.The shape of filtering blebs and the ocular anterior segment response were examined under microscope daily.The animals were sacrificed by excessive anesthesia on day 7 and 14 after operation,respectively,and the specimens were prepared of the eyes.Regular histopathological examination was performed to observe the collagen fibrosis change,and immunohistochemistry was carried out to assay the dynamic expression of HSP47 depended on the fibrosis.Percentage of positive cells for HSP47 were calculated.The use and care of the experimental animals complied with the Regulation for the Administration of Affair Concerning the Experimental Animals by State Science and Technology Committee.Results Diffuse subconjunctival blebs were seen postoperatively in t0 eyes after surgery with mild inflammatory response in ocular anterior segment.Hematoxylin-eosin staining showed that compared with the specimens of the normal control group,fibroblasts obviously proliferated and collagenous fibers increased in the specimens of operative group.The HSP47 protein was less expressed in cytoplasm of many fibroblasts and less vascular endothelial cells in the normal rat specimens.The expressing intensity of HSP47 protein was enhanced.However,the expression intensity was weaker in the postoperative 14-day group in comparison with the 7-day group.The percentage of positive cells for HSP47 in the filtering blebs 7 days after operation was (56.40±1.12)％,that in the normal control group was (13.70±0.74)％,with a significant difference between them (P=0.000).In the postoperative 14-day group,the percentage of positive cells for HSP47 reduced from (56.40± 1.12) ％ to (23.90±0.76) ％ (P =0.000),but it was still higher than that of the normal control group (P =0.000).Conclusions Expression intensity of HSP47 alters as the activity of fibroblasts in filtering bleb,suggesting that HSP47 plays a role in collagen-fibrosis following filtering surgery in SD rats.
ABSTRACT
BACKGROUND:In the perspective of traditional Chinese medicine, few studies have focused on the compound preparations though there are many investigations. The present study was undertaken to investigate the effect of Zhenwu Tang Granule on chronic pressure-overloaded left ventricular hypertrophy in rats. METHODS:The study was performed at the laboratory of Guangzhou Institute of Respiratory Disease. Male SD rats were divided randomly into 3 groups:sham operation group (n=8), operation group (n=15) and traditional Chinese medicine (TCM) group (n=15).The model of myocardial hypertrophy was made by gradually constricting the abdominal aorta. Sixteen weeks later, cardiac ultrasonography was performed in all groups in order to ascertain post-operational left ventricular (LV) hypertrophy. And Zhenwu Tang Granule was added at a dose of 12 g/kg in the mixed feedstuff for 8 weeks in the TCM group. In the 24th week, weight, structure as well as function of the heart in each group were measured by high-frequency ultrasonography, and Masson's staining was performed on the cardiac muscles. Meanwhile, total collagen volume fraction (CVF-T) and non-coronary vessel collagen volume fraction (CVF-NV) were analyzed. RESULTS:There was an increase in the weight of the heart in the operation group, with the left ventricule dominated (P<0.05). The heart was enlarged, with diastolic interventricular septal distance (IVSd) and left ventricular posterior wal distance (LVPWd) dominated (P<0.01).There was a significant decrease in the cardiac function (P<0.05). The weight (P<0.01) and volume of the heart decreased in the TCM group compared with the operation group, with IVSd and systolic left ventricular posterior wal dominated (P<0.01). And the cardiac function was improved (P<0.05). Significant interstitial and col agen hyperplasia was shown in the operation group based on pathological analysis, and various improvements were proved in the TCM group, i.e. there was a significant decrease in CVF-T and CVF-NV (P<0.01) compared with the operation group; but no difference (P>0.05) was found when compared with the pseudo-operation group. CONCLUSION:Zhenwu Tang Granule could reduce the weight and volume of the heart, improve the cardiac function, inhibit hyperplasia of collagen, and reverse myocardial hypertrophy in rats with pressure-overloaded left ventricular hypertrophy.
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Objective To study the effect of transplanted with human mesenchymal stem cells(MSCs) on the rat liver fibrosis in carbon tetrachloride(CCl-4)-induced hepatocirrhosis rats. Methods Wistar rats were divided into control(n=8) and hepatocirrhosis groups,and 24 hepatocirrhosis rats were made by subcutaneous injection with 60% CCl-4-peanut oil for 7 weeks,named 7-week hepatocirrhosis group(n=8).Other 7 weeks hepatocirrhosis rats were subdivided into MSC control group(n=8) and MSC transplanted group(n=8).After identification to MSCs with FACS calibur flow cytometry,MSCs were injected into the superior mesenteric vein for 3 weeks in MSC transplanted group.All of rats were killed at the beginning of 11-week.The clinical blood biochemical parameters from the inferior vena cava were examined.The survival of human MSCs in liver was observed by immunohistochemistry reaction of anti-hEGF-R.The degree of liver cirrhosis was evaluated by stains of Sirius red. Results Plasma ALP,ALB and CHO were abnormal to varying degree in 7-week hepatocirrhosis,MSC control and MSC transplanted groups.In 7 weeks hepatocirrhosis rats,a large amount of hyperplastic collagenous fiber was presented in liver tissue and the frame of the pseudolobuli was formed.These were similar to MSC control group.Only in MSC transplanted group,a few human EGF-R(+) cells were observed in livers.The amount of hyperplastic collagenous fiber in MSC transplanted group was significantly lower than those of the 7-week hepatocirrhosis and MSC control gruops.Conclusion Liver fibrosis could be improved obviously in hepatocirrhosis rats transplanted with human MSCs.These experimental results could probably provide an experimental evidence of treatment of liver cirrhosis.
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By using the LM,TEM and SEM the overall observations on both vertical and flat sections of 8 normal rabbit corneas have been made.For preparations of the corneal specimen,beside routine freezing and conventional ultrathin sectioning methods,the author used the personally designed methods:the freezing sectioning of the cornea flattened by pressing them onto a previousely prepared freezing plane(the F.P. method)in order to make flat sections of 8-10?m thick for the LM observation, and of 50-100?m thick for the SEM;The preparation of the cornea swollen with glacial acetic acid,sectioned by a freezing microtome for light microscopy——the GF method;The preparation of the cornea swollen and sectioned as mentioned above and by removing the ground substances in addition from the sections for scanning electron-microscopy——the GFR method;And the preparation of the cornea treated all the same as in the GFR method except the swelling procedure also for the scanning electronmicroscopy——the FR method.The ultrathin sections were mounted on the singl-hole grid in order to make continuous observation in a larger area.All LM,TEM and SEM specimens from the swollen corneas were compared with the unswollen control ones. As for the results,the findings by the 3 kinds of microscopes were consistent with each other.So were the findings of the flat sections with those of the vertical ones.In summary,the steoreo structure of the fibers of the corneal stroma is characterized by:(1)The appearance of the collagenous fibers is usually leaf-like. The fibers both criss-cross and anastomose with each other forming a network and they don't seem to lie parallel to one another,much less to form in such a way a sheet equal in width to the diameter of the cornea.The network with the thickness of about 4-9?m looks like a small board in an area of some 500?m around,called microboard(MB);(2)The MBs are connected with one another to form MB network——MBN;(3)The boundaries between the horizontally neigh- bouring MBs have no special structure,The place where a MB anastomoses with its upper or lower neighbours,can act as the margin of MBs;(4)The MB combines with the corneal cell into a cell-MB complex(CMC);(5)The so-called lamella or “collagenouS fibers”seen by the LM on the HE-stained vertical section is really the sections of the MBN;(6)The transmision electronmicrograph of the lamella shown in the literature is merely a small part of the MB;(7)Between the collagen- ous fibers there are a small number of very fine fibers in branched and interweaving form,which may be the elastic fiber. Above results explained why TEM observations in the past didn't accord with the LM ones both in thickness and in number of the lamellae.The 200-250 lamellae shown by the TEM correspond to the vertical sections of the collagenous fibers, while the 40-60 lamellae seen by the LM represent the vertical sections of the MBs.