Endothelin-1, an important mitogen of smooth muscle cells of spontaneously hypertensive rats

OBJECTIVE: To study the features of vascular smooth muscle cell (VSMC) proliferation induced by endothelin-1 (ET-1). METHODS: VSMCs of spontaneously hypertensive rats (SHR) and Wistar-Kyoto rats were cultured and treated with ET-1. Basic fibroblast growth factor (bFGF) gene expression was measured using both Northern blot and an enzyme-linked immunoassay. RESULTS: ET-1 resulted in an increase in bFGF transcripts at 8 - 24 h; bFGF levels were significantly higher in VSMCs treated with ET-1 than in those not treated. However, VSMCs growth responses in SHR and WKY were different. Smooth muscle cells of SHR were hyper-responsive to ET-1. Maximal bFGF mRNA levels were elevated 3.5-fold at 4 h of stimulation in WKY and 8-fold at 8h in SHR4. Moreover, the proliferation of VSMCs induced by ET-1 was inhibited by antisense phosphorothioate oligodeoxynucleotides (10 micromol/L AS-bFGF) but not sense bFGF oligomers at the same concentrations, being reduced by 80% in SHR and 40% in WKY vs control, respectively. Furthermore, the effect of AS-bFGF oligomers on SHR SMC proliferation is significantly greater than on WKY SMC proliferation. CONCLUSION: ET-1 may be required for exaggerated vascular growth responses in SHR and bFGF may be involved.

Influence of Valsartan on myocardial apoptosis in spontaneously hypertensive rats

OBJECTIVE: To explore the pathogenic changes of myocardial apoptosis in heart hypertrophy during hypertension and evaluate the anti-apoptosis effect of Valsartan. METHODS: Thirty spontaneously hypertensive rats (SHRs) were divided into two groups: 15 treated with Valsartan (20 mg x kg(-1) x d(-1)) (SHR + Valsartan group), the others with placebo (SHR + placebo group), with 15 normal Wistar rats as control. Systolic blood pressure was measured by the tail-cuff method. The observation period was from 8 to 16 weeks of age. Cardiac apoptosis was evaluated by a Terminal Deoxynucleotidyl Transferase-Mediated dUTP-biotin Nick End Labeling (TUNEL) assay. RESULTS: Mean blood pressure values were 127 +/- 2 mm Hg in controls, 163 +/- 6 mm Hg in the SHR + Valsartan group and 193 +/- 7 mm Hg in the SHR + placebo group at 16 weeks of age, whereas the blood pressure in 8-week-old SHR and Wistar rats were 175 +/- 3 mm Hg and 125 +/- 5 mm Hg, respectively. The ratio of the heart weight over body weight declined in Wistar (3.07 +/- 0.03 mg/g) and SHR + Valsartan groups (3.22 +/- 0.19 mg/g) compared with the SHR + placebo group (4.02 +/- 0.31 mg/g) (P

Endothelin receptor antagonist combined with a calcium channel blocker attenuates renal injury in spontaneous hypertensive rats with diabetes

OBJECTIVE: To investigate the effects of the mixed endothelin receptor antagonist, bosentan, combined with the long-acting calcium channel blocker, amlodipine, compared to the angiotensin-converting enzyme inhibitor, cilazapril, on the progressive renal injury in spontaneous hypertensive rats (SHR) with diabetes. METHODS: Diabetic hypertensive rats (SHR-DM) were induced by streptozotozin injected in male SHR (7-week-old),and divided into an untreated and three treated groups: 1) cilazapril treated group; 2) bosentan+amlodipine treated group; and 3) amlodipine treated group. Wistar Kyoto rats (WKY) and SHR rats served as normotensive and hypertensive control, respectively. The mean arterial blood pressure, renal function, endothelin and angiotensin II levels as well as the protein expression of renal extracellular matrix components and transforming growth factor (TGF)-beta1 were determined at the end of the 4th week. RESULTS: Mean arterial blood pressure significantly increased in SHR and SHR-DM rats compared to WKY rats. All the therapies reduced the blood pressure to normal levels. However, the enhanced urinary protein excretion, the decreased creatinine clearance as well as the increased plasma and intrarenal endothelin and angiotens in II levels were found in the untreated SHR-DM and prevented by treatment with bosentan+amlodipine and cilazapril. Similarly, these two kinds of therapies in SHR-DM abolished the overexpression of renal TGF-beta1 by Western blot analysis and reduced the accumulation of collagen type IV, laminin and fibronectin proteins by an immunochemical approach. Amlodipine monotherapy had no detectable effects on the above parameters. CONCLUSION: Bosentan combined with amlodipine can offer similar renoprotective effects on that of cilazapril and may be a potent therapy to attenuate renal injury by reducing renal protein levels of TGF-beta1 in diabetes with a hypertensive state.

Ventricular remodeling by Scutellarein treatment in spontaneously hypertensive rats

OBJECTIVE: To observe reversal of ventricular remodeling by the protein kinase C inhibitor Scutellarein in spontaneously hypertensive rats (SHRs). METHODS: Twelve SHRs were randomly divided into two groups. Scutellarein and saline (10 mg x kg(-1) x d(-1)) were given by intraperitoneal injection to two groups of rats separately. Systolic blood pressure (SBP) and ventricular weight index (LVW/BW, RVW/BW) were measured. A polarization microscope and an image analyzer system (IAS) were used to observe changes in cardiovascular structure and to count the content of cardiac muscle interstitial collagen. RESULTS: The pathologic changes in the left ventricle in the Scutellarein group rats (SHR(D)) improved to varying degrees, including hypertrophy of the cardiac muscle and collagen volume fraction. CONCLUSION: Scutellarein can reverse ventricular remodeling, improve myocardial stiffness and protect heart cardiac muscle.

Five novel monoclonal antibodies to thymic epithelial cell surface antigens in rats

OBJECTIVE: To establish monoclonal antibodies (mAbs) against thymic epithelial cells and study the function of epithelial cells during T-cell differentiation in the thymus. METHODS: Hybridomas secreting mAbs against thymic epithelial cells were derived by immunization of Balb/c mice with two thymic epithelial cell lines, TaD3 and FTE. The distribution of antigens recognized by these mAbs was detected by immunochemical staining and cytofluorographic analysis, and the molecular weight of the antigens by immunoblotting. RESULTS: Five specific monoclonal antibodies (mAb) were obtained. On the basis of their distribution in the thymus determined by immunochemical staining, mAb RE-4D8 was regarded as clusters of thymic epithelium staining (CTES) type IIA: mAb RE-12B2, which showed a unique distribution pattern only in the medulla, was CTES type V: mAb RE-5C6 was CTES type IV: mAb RE-6D6 might be CTES type IIB: and mAb RE-1D4 was classified as type V. The molecular weight (MW) of antigen RE-4D8, RE-6D6 and RE-12B2 were 120 kDa, 220 kDa and 35 kDa, respectively. Antigen RE-1D4 is a novel marker of cortical epithelium, several established thymic epithelial cell lines were classified and their original intrathymic locations were determined by these mAbs. Thymic cell lines, TuD3 and FTE were cortical phenotypes whereas TaD3 had a medullar phenotype. CONCLUSIONS: These mAbs clearly demonstrate the heterogeneity of the thymic epithelium; they could detect antigens not only in the cytoplasm but also on the surface of thymic epithelial cells. Our data suggest that these newly established mAbs may help elucidate the interaction between thymocytes and epithelial cells during T cell maturation.

DA到Lewis大鼠肝脏移植急性排斥模型的建立

目的：建立大鼠原位肝脏移植的急性排斥模型并观察其排斥反应的特点．方法：采用近交系雄性DA大鼠为供体，Lewis大鼠为受体，改良“二袖套”法建立肝脏移植模型30例．随机分为两组，对照组（n=15）术后不用免疫抑制剂，FK506处理组（n=15）术后1-7d用FK506 0．2mg／（kg·d）灌胃．术后4，7和10d分别随机处死3只，检测血清肝功能生化指标[天冬氨酸转氨酶（AST）、总胆红素（BILI）]以及肝脏病理组织学变化，每组各留6只观察生存期．结果：术后7和10d时对照组血清AST，BILI浓度均高于FK506处理组[AST：7d，（17．74±3．6）μkat/L vs（7．94±1．6）μkat/L，P〈0．05；10d，（21．94±4．3）μkat/L vs（4．74±0．7）μkat/L，P〈0．05．BILI：7d，（90．94±18．8）μmol／L vs（6．84±0．7）μmol／L，P〈0．05；10d，（111．54±23．3）μmol／L vs（15．54±3．2）μmol／L，P〈0．05]．术后7d，对照组移植物出现明显的排斥反应的病理学变化，10d时更为严重．FK506组术后7和10d病理学检查为不明确或轻度的排斥表现．对照组中位生存时间为12d，95％可信区间为（11．77-13．90）d（n=6，12d×4，14d×1，15d×1）．FK506处理组中位生存时间为78d，95％可信区间为（65．26-90．74）d（n=6，〉65 d×1，〉67d×1，76 d，78d，89d和90d），生存时间明显较对照组延长（P〈0．05）．结论：DA到Lewis大鼠品系组合可以建立稳定可靠的肝脏移植急性排斥模型；该模型的排斥反应的特点与临床相似，且可以为抗排斥药物FK506预防而达到长期存活．

PVG大鼠生化标记基因的测定

目的 对PVG大鼠进行生化标记基因的测定。方法 依据国家标准GB/T 14927.1—2008《实验动物近交系小鼠、大鼠生化标记检测法》测定,随机抽取4只PVG大鼠,取(肺、肾、小肠、睾丸)组织器官进行样品制备,用醋酸纤维薄膜电泳法检测11个生化位点(Hbb、Cs1、Alp1、Akp1、Esl、Es3、Es4、Es6、Es8、Es9和Esl0)。结果 PVG大鼠11个位点的基因型分别是Hbb为b型,Cs1为b型,Alp1为b型,Akp1为a型,Esl为a型,Es3为d型,Es4为b型,Es6为b型,Es8为b型,Es9为a型和Esl0为a型,且均为纯合。结论 PVG大鼠遗传生化标记结果符合近交系特征。

Effects of electroacupuncture at Taichong(LR 3) and Baihui(DU 20)on cardiac hypertrophy in rats with spontaneous hypertension

OBJECTIVE: To investigate the effects of electroacupuncture(EA) at Taichong(LR 3) and Baihui(DU 20)on myocardial hypertrophy in spontaneously hypertensive rats(SHRs).METHODS: Thirty-six SHRs were randomly assigned to model, EA, and Losartan groups, with twelve rats per group. Twelve Wistar Kyoto rats were selected as the normal control group. Systolic blood pressure(SBP) and cardiac function were measured in all rats.Expression levels of factors associated with the phosphoinositide 3-kinase/protein kinase B/mammalian target of rapamycin(PI3K/Akt/mTOR) pathway were evaluated by Western blotting and real-time PCR.Pathological changes of the heart tissue were observed by hematoxylin-eosin staining.RESULTS: After treatment, enhanced SBP was significantly decreased in the EA and Losartan groups compared with the model group(P < 0.01). Echocardiographic and morphological analyses revealed that enhanced end-diastolic interventricular septal thickness and left ventricular posterior wall thickness, as well as ratio of left ventricular weight to body weight were markedly diminished in the EA and Losartan groups(P < 0.01 or P < 0.05), while reduced left ventricular end-diastolic dimension and left ventricular ejection fraction were significantly ameliorated(P < 0.01). Real-time PCR and western blotting analyses showed that the expression levels of PI3K,Akt, and mT OR in SHRs were significantly up-regulated by EA and Losartan(P < 0.01), while the expression levels of PTEN and ANP were down-regulated(P < 0.01).CONCLUSION: EA at Taichong(LR 3) and Baihui(DU20) inhibited the development of cardiac hypertrophy and improved the cardiac function in SHRs, possibly through regulation of the PI3K/Akt/mTOR signalling pathway.

Lewis大鼠骨髓间充质干细胞分离培养和鉴定

目的建立从Lewis大鼠骨髓中分离、培养间充质干细胞的方法,并对分离、培养的间充质干细胞进行鉴定.方法采用Percoll(1.073g/L)密度梯度分离和贴壁培养相结合的方法分离培养Lewis大鼠的骨髓间充质干细胞(MSC).MSC细胞鉴定:采用相差显微镜观察MSC的形态学特征;流式细胞仪检测细胞表面标志抗原CD29,CD90,CD34和CD45的表达率;苏丹Ⅳ染色检测MSC成脂细胞分化;碱性磷酸酶染色和茜素红染色检测MSC成骨分化.结果原代培养的MSC于24 h后贴壁,48～72 h形成集落,12～15 d可达到80%～90%融合.第1代细胞表面标记物CD29,CD90,CD34和CD45的阳性率分别为80.41%,66.27%,2.73%,0.74%.第3代细胞表面标记物CD29,CD90,CD34和CD45的阳性率分别为89.91%,88.17%,0.81%,0.17%.细胞周期分析显示,第5代细胞G0/G1期细胞占68.9%,S+G2+M期为31.1%.用α-MEM培养液培养的第6代细胞部分分化为成骨和成脂细胞.结论采用Percoll(1.073g/L)密度梯度分离和贴壁培养相结合的方法能够成功分离和培养大鼠的MSC.第5代MSC细胞保持分化潜能.

纯系LEW大鼠及裸小鼠膀胱灌注给药方式的探讨

目的探讨纯系LEW大鼠及裸小鼠膀胱灌注给药方法。方法显微镜下解剖雌性LEW大鼠及裸小鼠尿道并根据解剖特征改造套管针。通过插入尿道的套管针膀胱灌注墨水,同时观察插管及膀胱充盈和染色情况。持续观察小鼠和大鼠生存状况及尿道损伤情况。结果显微镜下可见雌性LEW大鼠及裸小鼠尿道与人存在明显差异。经尿道插管成功率高(100%),动物生活状况没有明显受到插管灌注影响。结论裸鼠膀胱灌注给药治疗是一种成功率高而可行的给药手段,动物膀胱灌注化疗模型可能为研究膀胱肿瘤治疗提供更为有效的工具。

Effect of Taichong(LR 3) acupuncture in spontaneously hypertensive rats

OBJECTIVE: To evaluate the effects of Taichong(LR3) acupuncture points(acupoints) on the expression of glucose transporter protein 1(GLUT1) in the hypothalamus of spontaneously hypertensive rats(SHRs) as measured by combined positron emission tomography and computed tomography(PETCT).METHODS: Spontaneously hypertensive rats(SHR)were divided into model, Taichong(LR 3) acupuncture, and sham groups. Additionally, Tokyo Wistar rats were used as the control group. Changes in blood pressure were recorded in different groups of rats before and after the corresponding treatment. Hematoxylin and eosin(HE) staining was used to study basic morphological changes, and immunohistochemistry was used to determine GLUT1 expression in the hypothalamus. Further,PET-CT was utilized to elucidate the antihypertensive mechanism after acupuncture at the Taichong(LR 3) acupoints.RESULTS: PET-CT indicated activation of the hypothalamus. Measurement of blood pressure showed that acupuncture at the Taichong(LR 3) acupoints lowered blood pressure. HE staining did not show any significant pathological changes, although differences in cell number were observed. Immunohistochemical analysis indicated a GLUT1 downregulation in the SHRs of the Taichong(LR 3) acupuncture group after the treatment.CONCLUSION: Acupuncture at Taichong(LR 3) acupoints lowered blood pressure in SHRs, with possible mechanisms being changes in cell number and GLUT1 expression in the hypothalamus.

Mechanism and effect of Shijueming (Concha Haliotidis) on serum calcium in spontaneously hypertensive rats

OBJECTIVE： To observe the impact of Shijueming （Concha Haliotidis） on spontaneously hypertensive rats via blood pressure, serum calcium, vascular smooth muscle membrane L-type calcium channel α1 C subunit （CaL-α1C）, plasma membrane calci- um-ATPase （PMCA） mRNA expression, and the L-type calcium channel in vascular smooth muscle cells. METHODS： Twelve-week-old male rats with sponta- neous hypertension were divided into three groups： a Shijueming （Concha Haliotidis） group （group 1）, a nifedipine group （group 2）, and a dis- tilled water group （group 3）. All were given a four-week treatment. Blood pressure and dissocia- tive serum calcium were examined before treat- ment. Blood pressure was taken every week during treatment. Atomic absorption spectrometry was used to examine dissociative serum calcium. Re-verse transcription-polymerase chain reaction was used to examine the expression of CaL-α1C and PM- CA1 mRNA. The patch clamp technique was used to examine the electrophysiological characteristics of the vascular smooth muscle cell calcium chan- nels. RESULTS： After treatment, blood pressure of the Shijueming （Concha Halioticlis） group lowered but not significantly （P〉0.05）. Blood pressure of the nifedipine group lowered significantly （P〈0.05）. Blood pressure of the distilled water group re- mained high. The concentration of serum calcium in the Shijueming （Concha Haliotidis） and the dis- tilled water groups lowered （P〈0.05）. Expression of CaL-α1C mRNA in the nifedipine group decreased compared with the distilled water group （P〈0.01）. There was the decreasing trend in the Shijueming （Concha Haliotidis） group, but it was not statistically significant. Shijueming （Concha Haliotidis） also had effects on the expression of PMCA1 mRNA but with- out statistical significance. However, there was a significant decreasing effect on vascular smooth muscle cell Ica-L flow. CONCLUSION： This study indicated that Shijuem- ing （Concha Haliotidis） could increase serum calci- um and decrease blood pressure. It may work by in- fluencing calcium channels, expression of PMCA1 mRNA, and regulating ion calcium channels and calcium-ATPase.

近交系LEWIS→BN大鼠原位肝移植急性排斥模型的建立

目的建立近交系LEWIS→BN大鼠原位肝移植急性排斥模型,分析手术成功率的影响因素及该模型的稳定性,并总结该模型区别于常规使用的SD、Wistar等封闭群大鼠间原位肝移植的特点。方法实验组选择近交系雄性LEWIS及BN大鼠各30只分别作为供、受体,对照组选择雄性BN大鼠各9只作为供、受体。采用Kamada"二袖套"法实施原位肝移植术,不吻合肝动脉;于术后3,5,7,9,11,13,15d处死受体获取肝脏组织,用中性甲醛固定后制作石蜡切片进行HE染色以判定急性排斥的程度。结果原位肝移植术成功率约为74%,导致手术失败的原因依次为:肝上下腔静脉出血,门静脉出血,麻醉意外和其他;LEWIS→BN组出现不同程度的排斥现象,而BN→BN组则无排斥现象。与封闭群大鼠肝移植比较,该模型具有自身特点,即在排斥出现的时间、程度和结果转归上表现并非完全一致,然而所有受体均出现了排斥现象。结论大鼠肝移植是目前研究肝移植理想模型,本研究采用Kamada的"二袖套"法成功建立了大鼠肝移植急性排斥模型。

DA→LEW大鼠肝脏急性排斥反应模型的技术改进与评价

目的:建立大鼠肝脏移植急性排斥反应模型并改进其手术技巧,对所建模型进行评价。方法:采用近交系雄性DA大鼠60只,Lewis大鼠120只,分为两组,其中A组30例:Lewis大鼠为供体(n=30),Lewis大鼠为受体;B组60例:DA大鼠为供体(n=60),Lewis大鼠为受体。采用改良“二袖套”法建立肝脏移植模型,观察手术成功率、平均存活时间、术后第1、3、5、7和10 d各时相点血清丙氨酸氨基转移酶(ALT)、总胆红素(TBIL)水平以及移植肝脏的病理学变化。结果:手术成功率为93.3%(84/90);总耗时(90.0±15.0)min;无肝期时间(14.2±2.6)min;平均存活时间A组超过100d,B组(16.2±1.4)d;术后第5 d B组血清ALT和TBIL升高,与A组比较差异非常显著(P<0.01);B组移植肝脏病理检查有明显的排斥反应,而A组没有。结论:DA→LEW(Lewis)大鼠的品系组合是稳定的肝脏移植急性排斥反应模型;通过改进手术技巧提高了手术成功率,为进一步研究提供技术保障。

Proliferation of aortic smooth muscle cells and renin-angiotensin system in SHR rats

目的：探讨ＳＨＲ大鼠主动脉平滑肌细胞（ＡＳＭＣ）异常增殖和肾素血管紧张素系统（ＲＡＳ）的关系．方法：测定血管紧张素Ｉ（Ａｎｇ）、卡托普利（Ｃａｐ）、沙拉新（Ｓａｒ）对培养的ＳＨＲ、ＷＫＹＡＳＭＣ增殖和Ａｎｇ、血管紧张素转化酶（ＡＣＥ）的影响．结果：Ａｎｇ在２％血清培养基中可刺激ＳＨＲＡＳＭＣ增生．ＳＨＲＡＳＭＣ分裂增殖能力比ＷＫＹ强，ＳＨＲＡＳＭＣＲＡＳ处于高功能状态．Ｃａｐ长期（４周）干预显著抑制ＳＨＲＡＳＭＣ异常增殖和Ａｎｇ、ＡＣＥ活性，Ｓａｒ长期干预同样抑制ＳＨＲＡＳＭＣ的增殖和ＡＣＥ活性，但Ａｎｇ水平反而升高．Ｃａｐ短期（２４小时）干预不影响两种大鼠ＡＳＭＣＲＡＳ．结论：Ｃａｐ和Ｓａｒ长期干预通过减少ＳＨＲＡＳＭＣＡｎｇ生成或阻断Ａｎｇ和特异受体结合，抑制其异常增殖．

Upregulation of the angiotensin-converting enzyme 2-angiotensin-(1-7)-Mas receptor axis by a combination of Yinyanghuo(Herba Epimedii Brevicornus) and Cheqianzi(Semen Plantaginis) improves erectile function in spontaneously hypertensive rats

OBJECTIVE:To evaluate the effects of a combination of Yinyanghuo(Herba Epimedii Brevicornus)(HEB)and Cheqianzi(Semen Plantaginis)(SP)on erectile dysfunction caused by essential hypertension in spontaneously hypertensive rats(SHRs),and to elucidate the role of the angiotensin-converting enzyme 2-angiotensin-(1-7)-Mas receptor(ACE2/Ang[1-7]/Mas receptor)axis in this process.METHODS:A total of 24 SHRs were randomly assigned to three groups:SHR-control,low-dose(12.5 g/kg)and high-dose(25 g/kg)HEB+SP(HEBSP).Eight Wistar-Kyoto rats were used as normal controls.HEBSP was administered by oral gavage for 28 d.Erectile function was measured once a week using the Heaton test.After 4 weeks of treatment,the corpus cavernosum was harvested from each rat to measure nitric oxide(NO),nitric oxide synthase(e NOS)and Ang(1-7)levels,as well as ACE2,Mas receptor and neuronal nitric oxide synthase(n NOS)protein expression.RESULTS:After 4 weeks of treatment,HEBSP significantly increased erectile function in the treated group compared with SHR-control group(P<0.01).Additionally,HEBSP treatment significantly increased cavernosal levels of Ang(1-7),e NOS and NO.Moreover,HEBSP significantly elevated the expression levels of ACE2,Mas receptor and n NOS.These beneficial effects were elevated in the high-dose HEBSP group.CONCLUSION:HEBSP improved erectile function in SHRs by upregulating the ACE2/Ang(1-7)/Mas receptor axis,e NOS and n NOS pathways.

Lewis型至Brown Norway型大鼠肝移植急性排斥反应模型的建立

目的:建立Lewis(LEN)型到Brown Norway(BN)型大鼠肝移植急性排斥反应模型并观察其排斥反应特点。方法:采用近交系雄性LEW大鼠为供体,BN大鼠为受体,改良"二袖套"法建立模型36例,术后第3、7和10天分别随机解剖8只受体大鼠,检测血清肝功能生化指标,观察肝脏病理组织学变化,剩余受体大鼠观察生存时间。结果:受体大鼠术后第3、7和10天的血清总胆红素分别为(8.75±1.98)μmol/L、(21.12±2.03)μmol/L和(53.63±4.24)μmol/L,丙氨酸转氨酶(ALT)分别为(291.88±6.83)U/L、(462.33±16.98)U/L和(906.25±68.55)U/L,两者术后均逐渐升高;受体大鼠术后第3天光镜下汇管区见混合性炎细胞浸润和静脉内皮炎,伴部分肝实质变性,排斥活动指数(RAI)为4～5;术后第7天排斥反应加重,并出现小胆管增生,RAI为5～7;术后第10天排斥反应最严重,出现明显汇管区结构破坏,肝小叶消失,大量淋巴细胞和单核细胞浸润,静脉内皮炎明显,肝实质桥接坏死,RAI为7～9分;受体大鼠的临床表现及生存情况与病理变化一致。结论:LEW到BN大鼠组合有助于建立稳定可靠的肝移植急性排斥模型,且该模型的排斥反应特点与临床相似。

对DA-Lewis大鼠肝移植模型建立的探讨

目的探讨近交系大鼠肝移植的手术技巧，建立稳定的急性排斥模型。方法DA大鼠作为供体，雄性Lewis大鼠作为受体，采用改良后的Kamada“二袖套管法”，建立大鼠原位肝移植模型。观察术后存活情况，并对其生存时间进行统计分析。结果60只大鼠肝移植手术中，供体手术时间（30±8）rain，修肝时间（15±2）min，受体手术时间（60±7）min，无肝期时间（21±3）min，手术成功率88．3％。术后4～5d大鼠开始出现黄疸，精神差，睁眼困难，反应迟钝，并于术后11d内全部死亡。结论根据近交系大鼠自身结构特点，对大鼠原位肝移植模型建立方法进行了改进，取得了一定的成效。DA—Lewis为稳定、可靠的大鼠肝移植急性排斥模型，是研究肝移植排斥反应及免疫耐受的理想动物模型。

建立6种近交系大鼠遗传质量检测及品系鉴别的单核苷酸多态性组合

目的 筛选出均匀分布在大鼠染色体上的单核苷酸多态性(SNP)位点,建立常用近交系大鼠遗传质量检测及品系鉴别方法。方法 在文献中获得SNP共141个,利用Ensembl数据库中获取的信息筛选出在BN、F344、GK、LEW、SHR和WKY 6种大鼠品系中呈现多态性的SNP位点,并从中优化出均匀分布各染色体且包含品系特异性SNP的最佳位点组合。通过PCR扩增技术和Sanger测序方法对组合位点用于常用近交系大鼠遗传质量检测和品系鉴别的效果进行验证。结果 优化出了在各品系内表现单态性、品系间呈多态性的40个SNP标记。其中包含可以用于品系鉴别的特异性位点:4个BN大鼠特异性位点,3个F344大鼠特异性位点,2个GK大鼠特异性位点,2个LEW大鼠特异性位点和1个SHR大鼠特异性位点。分别利用BN、F344、LEW、SHR和WKY大鼠的DNA混合样本对优化出的40个位点进行验证,结果与网站数据一致。结论 成功建立了常用近交系大鼠遗传质量检测及品系鉴别的SNP组合。

Tong-xin-luo capsule inhibits left ventricular remodeling in spontaneously hypertensive rats by enhancing PPAR-γ expression and suppressing NF-κB activity

Background Tong-xin-luo capsule （TXL）, used as a traditional Chinese herb, offeres a therapeutic potential for treatment of cardiovascular diseases. It has been shown to exert a variety of pharmacological effects, including antihypertensive effects, and is able to improve ventricular remodeling. However, the mechanisms of its action are not completely understood. The aim of this study was to evaluate the molecular mechanisms of Tong-xin-luo capsule on left ventricular remodeling in spontaneously hypertensive rats （SHR）. Methods Sixteen eight-week-old SHRs were randomized into an SHR group （n=8） and a TXL group （n=8） that were given Tong-xin-luo capsule （1.5 mg·kg^-1·d^-1）. Eight Wistar Kyoto （WKY） rats fed with 0.9% NaCl served as the control group （WKY group）. Systolic blood pressure （BP）, body weight and heart rate were monitored once every two weeks. Ventricular remodeling was detected by histopathological examination. Nuclear factor kappa B P65 （NF-κB P65） and peroxisome proliferators activated receptor y （PPAR-γ） protein and phosphorylated inhibitor kappa a （IκBα） protein were detected by immunohistochemistry and western blot respectively. The physical interaction of the P65-P50 heterodimer with IκBα and NF-κB were measured by co-immunoprecipitation. PPAR-γ mRNA, collagen Ⅰ mRNA and collagen Ⅲ mHNA were measured by real-time PCR.Results TXL inhibited NF-κB P65 expression and ventricular remodeling and suppressed the activation of NF-κB compared with the SHR group （P〈0.01, P〈0.05）. TXL reduced IκBα phosphorylation, increased expression of PPAR-γ protein and enhanced the physical interaction of the P65-P50 heterodimer with IκBα. The mRNA expression of PPAR-γ was enhanced but the mRNA expression of collagen Ⅰ mRNA and collagen Ⅲ mRNA were suppressed by TXL. Conclusions In spontaneously hypertensive rats, TXL could inhibit ventricular remodeling induced by hypertension, and the inhibitory effect might be associated with the process of TXL increasing the expression of PPAR-γ that could result in the inhibition of the activation of NF-κB.