Adenovirus-mediated P311 ameliorates renal fibrosis through inhibition of epithelial-mesenchymal transition via TGF-ß1-Smad-ILK pathway in unilateral ureteral obstruction rats.

Epithelial-mesenchymal transition (EMT) is a critical step and key factor during renal fibrosis. Preventing renal tubular EMT is important for delaying the progression of chronic kidney disease (CKD). P311, a highly conserved 8-kDa intracellular protein, has been indicated as an important factor in myofibroblast transformation and in the progression of fibrosis. However, the related studies on P311 on renal fibrosis are limited and the mechanisms of P311 in the progression of renal tubulointerstitial fibrosis remain largely unknown. In the present study, we examined the effect of P311 on transforming growth factor-ß1 (TGF-ß1)-mediated EMT in a rat model of unilateral ureteral occlusion (UUO) renal fibrosis. The recombinant adenovirus p311 (also called Ad-P311) was constructed and transferred it into UUO rats, the preventive effect and possible mechanism of P311 on TGF-ß1-mediated EMT were explored. The UUO model was established successfully and Ad-P311 was administered into UUO rats each week for 4 weeks, then the serum levels of Cr, blood urea nitrogen (BUN) and albumin (ALB) were evaluated. H&E staining and Masson staining were performed to observe the pathological changes of kidneys. Immunohistochemical staining and western blot analysis were used to examine the EMT markers [E-cadherin and α-smooth muscle actin (α-SMA)], and signal transducers (p-Smad2/3 and Smad7). Integrin linked kinase (ILK) as a keyintracellular mediator that controls TGF-ß1-mediated-EMT was also assayed by western blot analysis. The results showed that P311 could alleviate renal tubular damage and interstitial fibrosis improving Cr, BUN and ALB serum levels in UUO kidneys. Furthermore, P311 attenuated TGF-ß1-mediated EMT through Smad-ILK signaling pathway with an increase in α-SMA, pSmad2/3 and ILK expression, and a decrease in E-cadherin and Smad7 expression in UUO kidneys. In conclusion, P311 may be involved in the pathogenesis of renal fibrosis by blocking TGF-ß1-mediated EMT via TGF-ß1-Smad-ILK pathway in UUO kidneys. P311 may be a novel target for the control of renal fibrosis and the progression of CKD.

Knockdown of ubiquitin­specific peptidase 39 inhibits the malignant progression of human renal cell carcinoma.

Ubiquitin specific peptidase 39 (USP39) serves important roles in mRNA processing and is involved in tumorigenesis of multiple solid malignancies. However, the influence and underlying mechanism of USP39 on human renal cell carcinomas (RCC) remain to be elucidated. The current study investigated the functional roles of USP39 in human RCC cell lines. siRNA­mediated RNA interference was used to downregulate USP39 in RCC cells. CCK­8, wound healing and invasion assays were performed to assess the proliferative ability and metastatic potential. The cell cycle distribution and apoptosis were evaluated by flow cytometry. The activity of signaling pathways and the expression of cell cycle­related proteins were detected by western blot analysis. The siRNA­directed RNA interference targeting USP39 could effectively downregulate the expression level of USP39 in two RCC cell lines. Depletion of USP39 by siRNA significantly suppressed cell growth and decreased invasive capacity of RCC cells. Silencing of USP39 induced cell apoptosis and cell cycle arrest at G2/M phase. Additionally, the expression levels of apoptotic and G2/M phase­related proteins were notably decreased following depletion of USP39. Mechanistically, downregulation of USP39 blocked the activation of Akt and extracellular signal regulated kinase signaling pathways in RCC cells. These findings indicate that USP39 may serve as an oncogenic factor in RCC and could be a potential therapeutic candidate for human RCCs.

[A literature review on Chinese medicine syndrome and syndrome elements of chronic fatigue syndrome].

OBJECTIVE: To research the distribution characteristics of Chinese medicine (CM) syndrome and syndrome elements of chronic fatigue syndrome (CFS) by analyzing literature in recent 20 years. METHODS: Relevant literature on treating CFS by syndrome differentiation of CM at home were retrieved by computer and manual ways. Database were established by using EpiData 3.1 to conduct frequency analysis of syndrome and syndrome elements. RESULTS: The most common clinical syndromes were Xin-Pi deficiency syndrome, Gan stagnation Pi deficiency syndrome, Gan-Shen yin deficiency syndrome, Gan qi stagnation syndrome, and Pi-Wei qi deficiency syndrome. Disease locations were sequenced as Pi, Gan, Shen, and Xin. The clinical pathogenesis of CFS was characterized by deficiency of vital energy, complicated with intermingled excess and deficiency. Asthenia of healthy energy was mainly manifested as qi deficiency, blood deficiency, and yin deficiency, while excess of sthenia was mainly manifested as qi stagnation, phlegm dampness, and static blood. CONCLUSIONS: Research of CM syndrome starting from syndrome elements can better unify and standardize clinical syndrome differentiation. Results of literature analysis can provide reference for further studies.

[Effects of Shenqi Jiedu Decoction on expressions of transforming growth factor-beta1, smad2 and smad3 in renal tissues of rats with chronic renal failure induced by adenine].

OBJECTIVE: To investigate the effects of Shenqi Jiedu Decoction (SQJDD), a compound traditional Chinese herbal medicine, on expressions of transforming growth factor beta1 (TGF-beta1), smad2 and smad3 proteins in renal tissues of rats with chronic renal failure (CRF) induced by adenine, and to explore the mechanisms of SQJDD in treating CRF. METHODS: Ninety healthy Wistar male rats were randomly divided into six groups: normal group, untreated group, losartan group, low-dose SQJDD group, medium-dose SQJDD group, and high-dose SQJDD group. Rat model of CRF was established by feeding 2.5% adenine. Levels of plasma creatinine (Cr) and blood urea nitrogen (BUN) in different groups after model establishment were detected to estimate the renal function of rats with CRF. After eight-week treatment, levels of Cr, BUN, triacylglycerol (TAG), cholesterol (Chol) and albumin (ALB) in plasma were detected. Pathological changes of renal tissue were observed by HE staining. Protein expressions of TGF-beta1, Smad2 and Smad3 in renal tissues were detected by Western blotting. RESULTS: Compared with pretreatment, levels of Cr and BUN in the losartan group and the SQJDD groups were markedly decreased (P<0.01). After treatment, levels of Cr, BUN and Chol in all treated groups were lower than those in the untreated group (P<0.01, P<0.05). Levels of ALB in all treated groups were higher than that in the untreated group (P<0.01). Pathological lesions of renal tissues in all treated groups were lessened as compared with the untreated group (P<0.01). The Western blotting results showed that the protein expressions of TGF-beta1, Smad2 and Smad3 in renal tissues in all treated groups were lower than those in the untreated group (P<0.01). CONCLUSION: SQJDD can delay the progress of CRF by inhibiting TGF-beta1/Smads signal transduction pathway.