Role of miR-219 and TGFBR2 in pathogenesis of renal fibrosis

AIM:To study the role of microRNA-219 (miR-219) in regulation of transforming growth factor-β receptor type 2 (TGFBR2) in renal fibrosis. METHODS:The renal fibrosis patients (n=70) were selected in this stu-dy, and 20 cases of healthy people were selected as control group. RT-qPCR was used to detect the expression of miR-219 in the serum of the patients with renal fibrosis and control group, and the expression of miR-219 in NRK49F cells after stimulation with angiotensin Ⅱ(AngⅡ) was detected. The protein expression of α-smooth muscle actin (α-SMA) in the NRK49F cells transfected with miR-219 mimics after stimulation with AngⅡ was determined by Western blot. The potential target gene TGFBR2 of miR-219 was screened and verified by the method of luciferase reporter gene. RT-qPCR and Western blot were used to detected the effect of miR-219 mimics on the expression of TGFBR2 at mRNA and protein levels, and the mRNA expression of α-SMA, connective tissue growth factor (CTGF), type I collagen α1 (COL1A1) and COL3A1 in the NRK49F cells was also detected, respectively. The unilateral ureteral occlusion (UUO) mouse model was established and the expression of miR-219 in the renal tissue was monitored. The morphological change of renal fibrosis was observed in the UUO mice after injection of miR-219, and the mRNA expression levels of COL1A1 and COL3A1 were detected. RESULTS:The expression level of miR-219 in the patients with renal fibrosis was significantly lower than that in control group, and the expression of miR-219 in the UUO mice was decreased significantly (P<0.01). The expression level of miR-219 was significantly decreased in the NRK49F cells after AngⅡ stimulation, and miR-219 mimics inhibited the protein expression of α-SMA(P<0.01). miR-219 mimics had a targeted regulatory effect on TGFBR2 gene, which inhibited the mRNA and protein expression of TGFBR2. miR-219 mimics inhibited the mRNA expression of α-SMA, CTGF, COL1A1 and COL3A1. miR-219 also down-regulated the mRNA expression of COL1A1 and COL3A1 in the UUO mice and inhibited the process of renal fibrosis. CONCLUSION:miR-219 inhibits the development of renal fibrosis by inhibiting the expression of TGFBR2, which may become a new target for the diagnosis and treatment of renal fibrosis.     

Effect of miR-29b-mediated TGF-β/Smad signaling pathway on progression of hepatic fibrosis in rats

AIM: To investigate the role of microRNA-29b (miR-29b)-mediated TGF-β/Smad signaling pathway in the activation of hepatic stellate cells (HSC) and its effect on the progression of hepatic fibrosis in rats.METHODS: Hepatic liver fibrosis rat model was established, and its HSC were isolated. Normal rat HSC were also obtained and identified in vitro. RT-qPCR and Western blot were used to detect the alterations of miR-29b, TGF-β/Smad signaling pathway-related proteins and liver fibrosis marker proteins in the acquired cells. Finally, the direct targeting binding of miR-29b to TGF-β1 was identified by dual-luciferase reporter assay system.RESULTS: With the activation of HSC, the expression of miR-29b gradually decreased (P<0.01), while the expression of collagen type I and α-smooth muscle actin gradually increased (P<0.01). At the same time, the expression of Smad2/3/4 was significantly increased, and the expression of Smad7 was significantly decreased (P<0.01). Dual-luciferase reporter assay showed that miR-29b bound directly to "UCUCUCCGU" in the 3'UTR of TGF-β1, indicating that TGF-β1 was a downstream target gene of miR-29b.CONCLUSION: miR-29b may be involved in the inhibition of HSC activation and migration, thereby inhibiting the process of liver fibrosis. The biological function of miR-29b may be through the direct targeting of TGF-β1, thus regulating and inhibiting the TGF-β/Smad signaling pathway.     

Notch1 promotes renal tubulointerstitial fibrosis in renal tissues of diabetic mice by inhibiting autophagy through Akt/mTOR signaling pathway

AIM: To observe the changes of Notch1 expression and autophagy in the renal tissues of diabetic mice, and to explore the regulatory effect of Notch1 on tubulointerstitial fibrosis by inhibiting autophagy in diabetic nephro-pathy. METHODS: The mice were randomly divided into normal control group (db/m mice) and diabetes group (db/db mice), with 8 rats in each group. After 12 weeks of feeding, the mice were sacrificed and the corresponding biochemical indexes were measured. The protein expression of Notch1 in the renal tubular epithelial cells was observed by immunohistochemical staining. The protein levels of Notch1, PTEN, p-Akt (Thr308), Akt, p-mTOR (Ser2448), mTOR, LC3, P62, collagen type Ⅰ (Col-Ⅰ) and collagen type Ⅲ (Col-Ⅲ) were determined by Western blot. RESULTS: Compared with the db/m mice, the blood glucose, glycosylated hemoglobin, serum creatinine, triglyceride and total cholesterol were increased in the db/db mice (P<0.01). Renal tubular epithelial cell vacuolar degeneration, renal tubular expansion and interstitial inflammatory cell infiltration in db/db mouse renal tissues with HE staining were observed. The images of Masson staining showed collagenous fiber-like substance deposition in the glomerular capillaries and renal interstitium, and disarrangement of tubular structure in the renal tissues of db/db mice. The protein expression levels of PTEN and LC3-Ⅱ were decreased (P<0.01 or P<0.05), while the protein levels of Notch1, P62, p-mTOR (Ser2448), p-Akt (Thr308), Col-I and Col-III were increased in the db/db mice as compared with the db/m mice (P<0.01). However, no significant change of total mTOR and Akt proteins between the 2 groups was found. CONCLUSION: Notch1 protein expression was increased, PTEN expression was significantly reduced, Akt/mTOR pathway was activated, autophagy was inhibited, and fibrosis was aggravated in the renal tissues of the diabetic mice.     

河豚的食用安全性及营养价值研究进展

阐述了河豚食用安全性,以及国内外对河豚鱼肉、肝脏和精巢营养价值的研究现状,以期为养殖低毒或无毒的河豚、科学地利用河豚资源及开放河豚市场提供参考。     

饲粮中添加啤酒花渣对肉仔鸡肝和肾功能的影响

[目的]探讨啤酒花渣的饲用价值。[方法]各试验组肉仔鸡饲粮中分别添加0、4％、8％、12％、16％和20％啤酒花渣,研究啤酒花渣对肉仔鸡肝和肾功能的影响。[结果]各试验组肉仔鸡肝脏超氧化物歧化酶（SOD）、过氧化氢酶（CAT）、总抗氧化能力（T-AOC）、谷胱甘肽过氧化物酶（GSH—Px）和丙二醛（MDA）差异均不显著（P〉0．05）。各试验组肉仔鸡肝脏和肾脏谷丙转氨酶（GPT）、谷草转氨酶（GOT）、乳酸脱氢酶（LDH）、胆碱酯酶（CHE）、醛缩酶（ALD）差异均不显著（P〉0．05）。[结论]饲粮中添加啤酒花渣对肉仔鸡肝和肾功能无不良影响。     

红树植物老鼠簕的研究进展

从多糖成分、甾醇成分、生物碱A等方面对近年来红树植物老鼠簕的化学成分研究及在护肝方面临床药理研究进行综述,为进一步开发利用老鼠簕提供依据。     

长白猪肝脏、肾脏和淋巴结的显微形态及糖原PAS反应研究

[目的]对商品长白猪的组织形态学进行研究,了解商品长白猪宰前的生理状态。[方法]采集长白猪的肝脏、肾脏和淋巴结3种组织器官并制成石蜡切片,采用H.E染色方法对其组织结构进行研究。[结果]肾脏和肝脏的组织结构形态正常,肾脏和肝脏的细胞核染色明显,淋巴结的皮质和髓质分界不清晰,周围组织没有明显的淋巴索,淋巴窦较小。糖原PAS染色结果表明肝脏中的糖原颗粒较少,分布不均匀,而肾脏中糖原或糖蛋白多集中于血管球和肾小囊壁上。[结论]该研究可为今后进一步的商品长白猪理论研究奠定基础。     

甘蔗汁对大鼠酒精性肝损伤的保护作用

[目的]通过建立大鼠酒精性肝损伤病理模型,探讨甘蔗汁对大鼠酒精性肝损伤的保护作用。[方法]取大鼠40只随机分为4组,正常组、低剂量组、高剂组量和模型组。采用白酒灌胃的方法建立大鼠酒精性肝病模式,给予甘蔗汁进行保护。测定肝组织中超氧化物歧化酶（SOD）活力、丙二醛（MDA）含量,并取肝组织制作石蜡切片,HE染色,普通光镜观察。[结果]甘蔗汁能够直接提高SOD活性、降低MDA含量,且低剂量（7.5 m l/kg）和高剂量（15 m l/kg）能够有效清除活性氧和自由基,明显改善肝组织脂肪样变。[结论]甘蔗汁能抑制酒精诱导脂质过氧化反应对肝组织的损伤,对酒精性肝损伤有明显的保护作用。     

雏鸭饲粮锰适宜水平的研究

采用玉米-豆饼型饲粮(含Mn19ppm),对210只1日龄北京雏鸭进行补锰试验,试验分7组,锰添加水平(试剂级MnSO_4·H_2O)按1～7组顺序依次为0,20,45,60,85,110,135ppm,试验期3周,研究雏鸭饲粮锰适宜水平。结果为,饲粮锰水平不影响雏鸭体增重,日采食量和饲料转化率(P>0.05);胫骨锰浓度随饲粮锰水平增加呈线性上升(P<0.01);肝、肾、胰、心组织锰浓度和肝脏含锰的SOD酶活性可与饲粮锰水平拟合为相应的二次曲线模式(P<0.001),上述软组织锰浓度和肝SOD酶活性达到平衡时,所需饲粮锰依次为110.05,100.00,122.91,113.99和116.00ppm。综合试验结果考虑,雏鸭饲粮锰适宜水平为110ppm。