| Objective:Chronic hydrocephalus is a disease which has the main features such asexcessive cerebrospinal fluid secretion, circulatory disorders, malabsorption,often secondary to intracranial hemorrhage and brain injury. There is noeffective precaution. The main treatment is cerebrospinal fluid shunt. But itoften failed because of complications such as infection and tube blocking.Until recently, numerous studies indicate that the key factors of chronichydrocephalus are the increase of apoptosis and extracellular matrix, whichcaused by the inflammatory responses. In this experiment, we will establish aintraventricular hemorrhage model of rat. We inject autologous arterial wholeblood, platelet-containing plasma and platelet-poor plasma into the lateralventricle of the different groups of rats, and then observe their muscle strength,masonic movement, and the area of ventricle and the expression of CTGFwithin brain tissue. Through this study, we can understand the role of differentblood components and CTGF in chronic hydrocephalus generation of rats.Methods：Choosing Wistar rats weight in180-220g, the number of male and femalewere equal. Before the start of the experiment, all rats were weighed andsubjected to three kinds of test thereby eliminate unqualified ones. Fail criteria:Screen test score≥1; Balance beam test score≥1; Climbing rope test score≥2.All qualified rats were subjected to the upper three kinds of test. All rats wereanesthetized by intraperitoneal injection of10%chloral hydrate. The blood isextracted from the femoral artery. All blood was centrifuged to obtainautologous arterial whole blood, platelet-containing plasma and platelet-poorplasma. All rats were divided into five groups：Blank group，Experimental control group, Autologous arterial whole blood group, Platelet-containingplasma group and Platelet-poor plasma group. The rats of blank group did notoperate. Experimental control group was injected into the ventricle with saline.The other groups were injected into the ventricle with the different bloodcomponent130μl. On the Seventh and28th day after operation, all rats wereweighed and subjected to three kinds of test. On the28th day after operation,all rats were anesthetized and perfusion fixed. The weight and volume of thebrain tissue samples were measured. Tissue sections were also stained withhematoxylin and eosin to observe histologic changes and calculate the area ofparacele. The expression of CTGF in the brain tissue samples were determinedby immunohistochemistry. Semi-quantitative analysis was carried out at equalpace.The data was expressed as mean±s and median of the indicated numberof samples studied. The data was assessed statistically using one-way analysisof variance (ANOVA) and nonparametric test. ANOVA was followed by LSDmultiple range tests using SPSS statistical software (version13.0) forcomparison between different treatment groups. Pearson correlation analysiswas adopted. Statistical significance was set at P <0.05.Results：1General observation1.1General stateThe rats of blank group show normally food intake, good mental state,shiny coat and flexible activities. The weight of rats gain with time.The rats of experimental control group showed piloerection, listlessness,reduced dietary water intake, reduced activity, after anesthesia recovery. Andthe rats restored on the second day.The rats of other groups showed piloerection, listlessness, reduced dietarywater intake, reduced activity and chills, in3days. And the rats restored on thefifth day. Compared with blank group, they showed less activities and the slowreaction.1.2Weight comparison (Table1) There was no significant difference in body weight among five groupsbefore modeling(P>0.05).But after7days, the weight of Autologous arterialwhole blood group, Platelet-containing plasma group, Platelet-poor plasmagroup and Experimental control group was lower than Blank groupremarkably (P<0.01).But there were no statistical differences among them(P>0.05). On the28th day after operation, there were no statistical differencesbetween Blank group and Experimental control group (P>0.05). The weight ofAutologous arterial whole blood group, Platelet-containing plasma group andPlatelet-poor plasma group was lower than Blank group remarkably (P<0.05).The weight of Autologous arterial whole blood group, Platelet-containingplasma group and Platelet-poor plasma group was lower than Experimentalcontrol group remarkably (P<0.05). The weight of Autologous arterial wholeblood group was lower than Platelet-containing plasma group andPlatelet-poor plasma group remarkably (P<0.05). There were no statisticaldifferences between Platelet-containing plasma group and Platelet-poorplasma group (P>0.05).2Behavioral comparison2.1Screen test score (Table2)The Screen Test score of rats has been compared by non-parametric tests.There were no statistical differences between Blank group, Experimentalcontrol group and Platelet-poor plasma group on the Screen Test score(P>0.05).The Screen Test score of Autologous arterial whole blood group,Platelet-containing plasma group was higher than other groups remarkably(P<0.01). The Screen Test score of Autologous arterial whole blood group washigher than Platelet-containing plasma group remarkably (P<0.01).2.2Balance beam experiment score (Table3)The Balance beam experiment score of rats has been compared bynon-parametric tests. There were no statistical differences between Blankgroup, Experimental control group and Platelet-poor plasma group on theBalance beam experiment score (P>0.05). The Balance beam experimentscore of Autologous arterial whole blood group, Platelet-containing plasma group was higher than other groups remarkably (P<0.01). The Balance beamexperiment score of Autologous arterial whole blood group was higher thanPlatelet-containing plasma group, but there was no statistical differencesbetween them (P>0.05).2.3Climbing rope test score (Table4)The Climbing rope test score of rats has been compared bynon-parametric tests. There were no statistical differences between Blankgroup and Experimental control group on the Climbing rope test score(P>0.05). The Climbing rope test score of Autologous arterial whole bloodgroup, Platelet-containing plasma group and Platelet-poor plasma group washigher than other groups remarkably (P<0.01). The Climbing rope test scoreof Autologous arterial whole blood group and Platelet-containing plasmagroup was higher than Platelet-poor plasma group remarkably (P<0.01). TheClimbing rope test score of Autologous arterial whole blood group was higherthan Platelet-containing plasma group, but there was no statistical differencesbetween them (P>0.05)3The weight, volume and density of brain tissue (Table5)Weight of brain tissue (g): Blank group1.915±0.026; Experimentalcontrol group1.902±0.028; Autologous arterial whole blood group1.853±0.033; Platelet-containing plasma group1.841±0.031; Platelet-poorplasma group1.863±0.023. There were no statistical differences betweenBlank group and Experimental control group on the weight of brain tissue(P>0.05). The weight of brain tissue of Autologous arterial whole blood group,Platelet-containing plasma group and Platelet-poor plasma group was higherthan other groups remarkably (P<0.05). But there were no statisticaldifferences among Autologous arterial whole blood group, Platelet-containingplasma group and Platelet-poor plasma group on the weight of braintissue(P>0.05). There was a significant positive correlation between theweight of body and the weight of the brain tissue [R=0.639(P<0.01)].The volume of the brain tissue (ml): Blank group1.971±0.027;Experimental control group1.953±0.029; Autologous arterial whole blood group1.905±0.037; Platelet-containing plasma group1.891±0.031; Platelet-poor plasma group1.915±0.020. There were no statistical differences betweenBlank group and Experimental control group on the volume of the brain tissue(P>0.05). The volume of the brain tissue of Autologous arterial whole bloodgroup, Platelet-containing plasma group and Platelet-poor plasma group washigher than other groups remarkably (P<0.05). But there were no statisticaldifferences among Autologous arterial whole blood group, Platelet-containingplasma group and Platelet-poor plasma group on the volume of the brain tissue(P>0.05).The density of brain tissue (g/ml): Blank group0.971±0.045; Experi-mental control group0.974±0.066; Autologous arterial whole blood group0.973±0.056; Platelet-containing plasma group0.977±0.056; Platelet-poorplasma group0.972±0.051. There were no statistical differences between thefive groups.4Ventricle width (Table6)The ventricle width (mm): Blank group0.511±0.058; Experimentalcontrol group0.507±0.051; Autologous arterial whole blood group1.200±0.045; Platelet-containing plasma group1.053±0.031; Platelet-poor plasmagroup0.589±0.106. There were no statistical differences between Blank groupand Experimental control group (P>0.05). The ventricle width of Autologousarterial whole blood group, Platelet-containing plasma group and Platelet-poorplasma group was higher than other groups remarkably (P<0.05). Theventricle width of Autologous arterial whole blood group was higher thanPlatelet-containing plasma group and Platelet-poor plasma group remarkably(P<0.05). The ventricle width of Platelet-containing plasma group was higherthan Platelet-poor plasma group remarkably (P<0.05).5The expression of CTGF (Table7)Percentage of CTGF positive cells in brain tissue (%):Blank group19.10±3.76; Experimental control group20.00±4.03; Autologous arterialwhole blood group50.70±9.27; Platelet-containing plasma group44.30±11.85;Platelet-poor plasma group39.40±4.25. There were no statistical differences between Blank group and Experimental control group on the percentage ofCTGF positive cells in brain tissue (P>0.05). The percentage of CTGFpositive cells in brain tissue of Autologous arterial whole blood group,Platelet-containing plasma group and Platelet-poor plasma group was higherthan other groups remarkably (P<0.01). The percentage of CTGF positivecells in brain tissue of Autologous arterial whole blood group was higher thanPlatelet-containing plasma group, but there were no statistical differencesbetween them (P>0.05). The percentage of CTGF positive cells in brain tissueof Platelet-containing plasma group was higher than Platelet-poor plasmagroup, but there were no statistical differences between them (P>0.05). Thepercentage of CTGF positive cells in brain tissue of Autologous arterial wholeblood group was higher than o Platelet-poor plasma group remarkably(P<0.05).Cell color intensity was graded by the method devised by Soslow[2]:0(Negative);1(Weakly positive);2(positive);3(Strong positive). The cellcolor intensity of rats has been compared by non-parametric tests. There wereno statistical differences between Blank group and Experimental control groupon the cell color intensity (P>0.05). The cell color intensity of Autologousarterial whole blood group, Platelet-containing plasma group and Platelet-poorplasma group was higher than other groups remarkably (P<0.01).The cellcolor intensity of Autologous arterial whole blood group and Platelet-containing plasma group was higher than Platelet-poor plasma groupremarkably (P<0.05). There were no statistical differences between Autolo-gous arterial whole blood group and Platelet-containing plasma group on thecell color intensity (P>0.05).The percentage of positive cells was graded by the method devised bySoslow[2]:0(0-1%);1(1-10%);2(10-50%);3(50-80%);4(80-100%). The IHSratings are calculated (IHS=Cell color intensity×Percentage of positive cells).The IHS ratings have been compared by non-parametric tests. There were nostatistical differences between Blank group and Experimental control group onthe IHS ratings (P>0.05). The IHS ratings of Autologous arterial whole blood group, Platelet-containing plasma group and Platelet-poor plasma group washigher than other groups remarkably (P<0.01). The IHS ratings of Autologousarterial whole blood group and Platelet-containing plasma group was higherthan Platelet-containing plasma group remarkably (P<0.01). There were nostatistical differences between Autologous arterial whole blood group andPlatelet-containing plasma group on the IHS ratings (P>0.05).Conclusion：1The ventricular expansion can be caused by the intraventricular inje-ction which with the following three blood components, autologous arterialwhole blood, platelet-containing plasma and platelet-poor plasma. Theventricle width of Autologous arterial whole blood group was higher thanPlatelet-containing plasma group and Platelet-poor plasma group remarkably(P<0.05). Platelets may play a role in the formation of hydrocephalus.2The abundantly expressed of CTGF in the brain tissue show that itplays a key role in the chronic hydrocephalus generation. The IHS ratings ofAutologous arterial whole blood group and Platelet-containing plasma groupwas higher than Platelet-containing plasma group remarkably (P<0.01).Platelets may play a role in the formation of hydrocephalus.3When the rat model of hydrocephalus is established, myodynamiadecrease and ataxia are possible. |
PDF Full Text Request