Effects of Lingguizhugan Decoction on α-SMA and collagen synthesis in rat myocardial fibroblasts induced by transforming growth factor-β_(1)

Objective:To explore the protective effect of Linggui Zhugan Decoction(LGZGD)medicated serum on myocardial fibrosis induced by transforming growth factor-β1(TGF-β1).Methods:Using enzyme digestion method,combined with differential adherence to isolate and culture Sprague-Dawley(SD)suckling mouse Cardiac fibroblasys(CFB)in vitro.Divided into:blank group,blank rat serum group,model group,and LGZGD medicated serum group(5%、10%、20%).Except for blank group and blank rat serum group,they were stimulated with 5 ng/ml TGF-β1 for 12 hours,and then then intervene with LGZGD medicated serum(5%、10%、20%)and continue to culture for 24 hours.Use immunofluorescence and Western blot(WB)to detect the expression ofα-smooth muscle actin(α-SMA),Enzyme-linked immunosorbent assay(ELISA)and WB to detect type Ⅰ collagen(Collagen Ⅰ),type Ⅰ collagen(Collagen Ⅲ)and fibronectin(FN)expression.Results:Compared with the blank group,the expressions of Collagen Ⅰ,Collagen Ⅲ,α-SMA and FN in the model group were significantly increased(P<0.01);Compared with the model group,the expressions of Collagen Ⅰ and Collagen Ⅲ in each concentration group of the experiment were significantly reduced(P<0.01);the expression ofα-SMA and FN were significantly reduced(P<0.01).Conclusions:LGZGD has an inhibitory effect on collagen synthesis and the expression ofα-SMA and FN,indicating that the anti-fibrosis effect of LGZGD is related to it.

Expression of Transforming Growth Factor β_(1) in Mesenchymal Stem Cells: Potential Utility in Molecular Tissue Engineering for Osteochondral Repair

The feasibility of using gene therapy to treat full-thickness articular cartilage defects was investigated with respect to the transfection and expression of exogenous transforming growth factor(TGF)-β_(1)genes in bone marrow-derived mesenchymal stem cells(MSCs)in vitro.The full-length rat TGF-β_(1)cDNA was transfected to MSCs mediated by lipofectamine and then selected with G418,a synthetic neomycin analog.The transient and stable expression of TGF-β_(1)by MSCs was detected by using immunohistochemical staining.The lipofectamine-mediated gene therapy efficiently transfected MSCs in vitro with the TGF-β_(1)gene causing a marked up-regulation in TGF-β_(1)expression as compared with the vector-transfected control groups,and the increased expression persisted for at least 4 weeks after selected with G418.It was suggested that bone marrow-derived MSCs were susceptible to in vitro lipofectamine mediated TGF-β_(1)gene transfer and that transgene expression persisted for at least 4 weeks.Having successfully combined the existing techniques of tissue engineering with the novel possibilities offered by modern gene transfer technology,an innovative concept,i.e.molecular tissue engineering,are put forward for the first time.As a new branch of tissue engineering,it represents both a new area and an important trend in research.Using this technique,we have a new powerful tool with which:(1)to modify the functional biology of articular tissue repair along defined pathways of growth and differentiation and(2)to affect a better repair of full-thickness articular cartilage defects that occur as a result of injury and osteoarthritis.

Study of Rat Osteoblasts Transfected by Transforming Growth Factorβ_(1)Gene

Summary:In order to investigate the effect of TGFβ_(1) gene transfer on the biological characteristics,the effects of gene transfer and supernatant of transfected osteoblasts on the proliferation and ALP activity of osteoblasts were detected by ^(3)H-TdR and MTT.Our results showed that TGFβ_(1) gene transfer had no effect on the biological characteristics and the activated supernatant of transfected osteoblasts stimulated proliferation and inhibited ALP activity of osteoblasts.TGFβ_(1) gene transfer could promote the expression of TGFβ_(1) and the biological characteristics of transfected osteoblasts were stable,which might be helpful for gene therapy of bone defects in vivo.

转化生长因子β_1基因修饰的间充质干细胞/仿生基质材料的体外复合培养

目的 探讨转化生长因子β_1(TGF-β_1)基因转染对间充质干细胞(MSCs)增殖分化的影响，评价涂覆多聚赖氨酸(PLYS)的聚DL乳酸(PDLLA) 仿生基质材料能否作为关节软骨组织工程学研究的支架材料。方法 将组织工程学与分子生物学有机结合，体外将具有多重生物学效应的TGF-β_1基因转入关节软骨组织工程首选种子细胞--间充质干细胞（MSCs），并与表面涂覆PLYS的PDLLA可降解三维多孔基质材料体外复合培养。以单纯空载体转染MSCs为对照，通过扫描电镜等方法观察种子细胞的粘附、增殖及分化等情况。结果 基质材料孔隙率为88%，其中孔径为150～200μm的有效孔占80%。所有细胞均能很好地粘附于基质材料上，其中TGF-β_1基因修饰的MSCs增殖分化活性明显优于对照组。结论 利用分子组织工程学原理可使TGF-β_1持续高效发挥作用，使提高关节软骨缺损的修复质量和远期疗效成为可能。涂覆PLYS的PDLLA仿生基质材料不仅具有良好的生物相容性和结构相容性，而且具有更好的表面相容性和生物学活性，在一定程度上解决了细胞-基质材料之间存在的界面不相容问题，是关节软骨缺损修复的良好基质材料。

Colonic expression of Runx3 protein and TGF-β_1 and their correlation in patients with irritable bowel syndrome

Objective:To investigate the role of Runx3 protein and TGF-β_1 in the pathogenesis of irritable bowel syndrome(IBS),as well as the correlation of these two proteins.Methods:Colonic tissue was collected from patients with IBS and normal persons.The colonic expression of Runx3 protein and TGF-β_1 was detected with immunohislochemistry method.Semi-quantitative analysis was used to evaluate the staining degree of these two proteins.Results:Compared with their counterparts,patients with IBS did not show any changes in the colonic expression of Runx3 protein and TGF-β_1(P>0.05).Interestingly,there was a significant correlation between Runx3 protein and TGF-β_1 in patients with IBS(P<0.05).Conclusions:The role of Runx3 protein and TGF-β_1 in the pathogenesis of IBS remains to be further studied.

转化生长因子β_l基因修饰成骨细胞复合仿生基质材料治疗骨缺损的研究

目的探讨转化生长因子β1(TGFβ1)基因修饰成骨细胞复合仿生基质材料修复鼠胫骨骨缺损的治疗效果。方法将TGFβ1基因转染成骨细胞后与涂覆多聚赖氨酸(PLYS)的聚DL乳酸(PDLLA)仿生基质材料复合,植入鼠胫骨骨缺损模型,术后摄X线片和组织学检查观察修复情况。结果实验组4周时X线片可见骨痂形成,组织学观察有类骨组织和新骨形成,成骨细胞贴附于基质材料表面。8周时缺损基本修复,新骨密度与自体骨接近;对照组4周时X线片未见骨痂形成,组织学观察没有类骨组织形成,基质材料表面成骨细胞贴附较少,材料腔隙中有大量淋巴细胞浸润。8周时植入材料基本吸收,为纤维组织替代。结论将分子生物学和组织工程学结合以修复骨缺损,能获得理想的治疗效果。

慢性肾小球疾病尿细胞因子改变及其与中医证型的关系

目的 探讨慢性肾小球疾病尿细胞因子改变及其与中医证型的关系。方法 对1 1 0例慢性肾小球疾病患者尿中白细胞介素 - 6 (IL - 6 )、转化生长因子 - β1 (TGF - β1 )及内皮素 - 1 (ET - 1 )进行了测定。结果 肾病综合征、慢性肾炎及慢性肾功能衰竭患者尿中IL - 6、TGF - β1 及ET - 1与健康对照组相比均有显著性差异 (P <0 .0 1或 0 .0 5 ) ,各型之间亦有一定的差异 (P <0 .0 1或 0 .0 5 )。中医证型阳虚型尿中TGF - β1 及ET - 1显著增高 (P均 <0 .0 1 ) ,气虚型及阴虚型均有尿IL - 6显著增高 (P <0 .0 1 ) ,但气虚型尿TGF - β1 亦增高 (P <0 .0 5 ) ;水湿证及湿热证尿IL - 6、TGF - β1 及ET - 1均显著增高 (P <0 .0 1或 0 .0 5 ) ,但TGF - β1 低于阳虚型 (P <0 .0 5 )。结论 不同临床类型及不同中医证型的肾小球疾病尿细胞因子的水平各有一定的特点 ,因此 ,尿细胞因子检查对中医辨证分型及了解肾内病理改变有一定的参考价值 ,从而为临床治疗提供相应的针对性。

胰腺癌转化生长因子-β_1和β-葡萄糖醛酸酶同步检测的临床意义

目的 探讨转化生长因子 β1(TGF - β1)和 β -葡萄糖醛酸酶 (β -GCD)在胰腺癌发生发展中的作用。方法 采用免疫组化ABC法同步检测胰腺癌和癌旁胰腺组织中TGF - β1和 β -GCD的表达。 结果 胰腺癌组织TGF - β1阳性细胞百分率 [(43 8± 5 2 ) % ]明显高于癌旁胰腺组织 [(2 8 7± 3 6 ) % ],P <0 0 1;癌细胞分化愈差或有淋巴结转移 ,TGF - β1过度表达愈多。胰腺癌β-GCD阳性细胞百分率 [(6 2 5± 4 1) % ]明显高于癌旁胰腺组织 [(33 5± 2 8) % ],P <0 0 1;β -GCD过度表达程度与癌细胞分化程度有关 ,但与淋巴结转移无关。TGF - β1和β-GCD在胰腺癌中表达呈一致性关系。 结论 胰腺癌的发生是多因素、多步骤、多基因变化的结果。TGF - β1和β-GCD的同步检测有助于胰腺癌恶性程度的判定和预后的评估。