Progress of heat shock protein 47 in glioma / 肿瘤研究与临床

Heat shock protein 47 (HSP47) is mainly involved in regulating collagen folding, secretion and maturation in the tumor microenvironment (TME) and is widely amplified in human cancers. HSP47 has been shown to be overexpressed in a variety of extracranial tumors. In glioma, the expression of HSP47 correlates with the grading of glioma and is involved in proliferation, invasion, angiogenesis, and immune regulation of glioma, regulates the TME of glioma, and promotes the survival of glioma stem cells, which may be related to the heterogeneity of glioma. This article reviews the progress of HSP47 in the development and progression of glioma, and discusses the significances of HSP47 in the proliferation, invasion, targeted therapy and immunotherapy of glioma.

Effects of HSP47 siRNA on biological behaviors of HTCF cells cultured <i>in vitro</i> and the expression level of TGF-β1 / 国际眼科杂志(Guoji Yanke Zazhi)

@#AIM: To explore the effects of heat shock protein 47(HSP47)siRNA on biological behaviors of human Tenon capsule fibroblasts(HTCF)cells cultured <i>in vitro</i> and the expression level of transforming growth factor-β1(TGF-β1).<p>METHODS: HTCF were cultured <i>in vitro</i> and divided into blank control group, empty vector group and transfection group. In transfection group, interfering siRNA sequences were designed and synthesized based on the HSP47 gene sequences, vectors were constructed and introduced into HTCF. The empty vector group was introduced with empty vectors. The expressions of HSP47 mRNA and protein in cells were detected by RT-PCR and Western blot. The proliferation, apoptosis, invasion and migration of cells were detected by clone formation assay, flow cytometry, Transwell method and scratch test. The expressions of proliferation, apoptosis, invasion and migration proteins, and TGF-β1 were detected by Western blot.<p>RESULTS: Compared with empty vector group, expression of HSP47 mRNA and protein, clone formation rate, cell healing rate, number of invasive cells, relative expression levels of Ki67, N-cadherin and TGF-β1 were significantly decreased in transfection group(<i>P</i><0.05), relative expression level of E-cadherin protein was significantly increased(<i>P</i><0.05), but there was no difference in apoptosis rate, and relative expression levels of Bcl-2 and Bax(<i>P</i>>0.05).<p>CONCLUSION: HSP47 siRNA can reduce proliferation, invasion and migration abilities of HTCF cells by inhibiting the expression of TGF-β1 protein, without significant effects on the apoptosis of HTCF cells.

Roles of transforming growth factor - β1 and heat shock protein 47 in progression of Schistosoma japonicum-induced hepatic fibrosis / 中国血吸虫病防治杂志

Objective To investigate the dynamic expression of transforming growth factor-β1 (TGF-β1) and heat shock protein 47 (HSP47) and explore their roles in the progression of hepatic fibrosis induced by Schistosoma japonicum infection. Methods Fifty female mice of the ICR strain were randomly divided into the infection group and the normal control group, of 25 mice in each group. Each mouse in the infection group was infected with 20 ± 1 cercariae of S. japonicum via the abdominal skin, while uninfected animals served as normal control. Five mice were sacrificed 4, 6, 8, 10 and 12 weeks post-infection and liver tissues were sampled. Serum HSP47 and TGF-β1 was determined using enzyme-linked immunosorbent assay (ELISA), and the pathological changes of liver specimens were observed with hematoxylin & eosin (HE) staining. In addition, the synthesis of alpha 1 chain of type I collagen (COL1A1) was measured using Masson staining, and the mRNA expression of TGF-β1, HSP47 and COL1A1 was determined using real-time fluorescent quantitative PCR (qPCR) assay. Results During the period of S. japonicum-induced hepatic fibrosis, the serum HSP47 and TGF-β1 levels and the mRNA expression of TGF - β1, HSP47 and COL1A1 gradually increased with the progression of hepatic fibrosis. The serum levels of HSP47 and TGF-β1 were (179.26 ± 29.87) pg/mL and (22.37 ± 5.21) ng/mL 6 weeks post-infection, respectively, which were significantly greater than those [(150.29 ± 34.91) pg/mL and (18.54 ± 7.78) ng/mL, respectively] in the normal control group (both P values < 0.05). In addition, the mRNA expression of HSP47, COL1A1 and TGF-β1 was (0.86 ± 0.04), (1.17 ± 0.06) and (0.64 ± 0.13) in mouse liver specimens, which was significantly higher than that (0.23 ± 0.03, 0.20 ± 0.02 and 0.38 ± 0.02) in the normal control group (all P values < 0.01). Conclusions The expression of TGF-β1 and HSP47 during the period of S. japonicum-induced hepatic fibrosis is consistent with the progression of the hepatic fibrosis, and exhibits the same tendency with type I collagen expression. HSP47 is a novel promising diagnosis marker and therapeutic target for S. japonicum-induced hepatic fibrosis.

Roles of transforming growth factor - β1 and heat shock protein 47 in progression of Schistosoma japonicum-induced hepatic fibrosis / 中国血吸虫病防治杂志

Objective To investigate the dynamic expression of transforming growth factor-β1 (TGF-β1) and heat shock protein 47 (HSP47) and explore their roles in the progression of hepatic fibrosis induced by Schistosoma japonicum infection. Methods Fifty female mice of the ICR strain were randomly divided into the infection group and the normal control group, of 25 mice in each group. Each mouse in the infection group was infected with 20 ± 1 cercariae of S. japonicum via the abdominal skin, while uninfected animals served as normal control. Five mice were sacrificed 4, 6, 8, 10 and 12 weeks post-infection and liver tissues were sampled. Serum HSP47 and TGF-β1 was determined using enzyme-linked immunosorbent assay (ELISA), and the pathological changes of liver specimens were observed with hematoxylin & eosin (HE) staining. In addition, the synthesis of alpha 1 chain of type I collagen (COL1A1) was measured using Masson staining, and the mRNA expression of TGF-β1, HSP47 and COL1A1 was determined using real-time fluorescent quantitative PCR (qPCR) assay. Results During the period of S. japonicum-induced hepatic fibrosis, the serum HSP47 and TGF-β1 levels and the mRNA expression of TGF - β1, HSP47 and COL1A1 gradually increased with the progression of hepatic fibrosis. The serum levels of HSP47 and TGF-β1 were (179.26 ± 29.87) pg/mL and (22.37 ± 5.21) ng/mL 6 weeks post-infection, respectively, which were significantly greater than those [(150.29 ± 34.91) pg/mL and (18.54 ± 7.78) ng/mL, respectively] in the normal control group (both P values < 0.05). In addition, the mRNA expression of HSP47, COL1A1 and TGF-β1 was (0.86 ± 0.04), (1.17 ± 0.06) and (0.64 ± 0.13) in mouse liver specimens, which was significantly higher than that (0.23 ± 0.03, 0.20 ± 0.02 and 0.38 ± 0.02) in the normal control group (all P values < 0.01). Conclusions The expression of TGF-β1 and HSP47 during the period of S. japonicum-induced hepatic fibrosis is consistent with the progression of the hepatic fibrosis, and exhibits the same tendency with type I collagen expression. HSP47 is a novel promising diagnosis marker and therapeutic target for S. japonicum-induced hepatic fibrosis.

Effects of 1,25-dihydroxyvitamin D3 on the expression of connective tissue growth factor and heat shock protein 47 in peritoneum fibrosis rats / 中华肾脏病杂志

Objective To investigate the expression of connective tissue growth factor (CTGF) and heat shock protein 47 (HSP47) in peritoneum fibrosis rats,and the mechanism of 1,25-dihydroxyvitamin D3 [1,25-(OH)2-VitD3] in inhibiting the peritoneum fibrosis.Methods Adult male Sprague-Dawley rats were randomly divided into 3 groups:control group (n=8),model group (n=8) and 1,25-dihydroxyvitamin D3 group (VitD3,n=8).The model of peritoneum fibrosis rats were induced by daily intraperitoneally injection of 15％ chlorhexidine gluconate (CHX) 0.2 ml/d with 0.1％ glucose for 4 weeks.Rats in VitD3 group were also treated with 1,25-(OH)2-VitD3 [i.p.6 ng· (100 g) 1 · d-1].Peritoneal transport function,renal function,peritoneum thickness and serum level of 25hydroxyvitamin D3 were detected.In vitro,primary cultured peritoneal mesothelial cells were divided into control group,high glucose group (HG,2.5％),CTGF siRNA intervention group (CTGF siRNA+HG),VitD3 intervention group (VitD3+HG) and combined intervention group (CTGF siRNA+VitD3+HG).Real-time PCR,Western blotting and immunofluorescence were applied to measure the expression of CTGF and HSP47,also ELISA was used to detect the protein level of FN in peritoneum and peritoneal mesothelial cells.Results Compared with control group,the peritoneal ultrafiltration in peritoneum fibrosis rats were significantly decreased (P ＜ 0.05),the absorbance level of peritoneal fibrosis,peritoneum thickness,the rate of dialysate urea nitrogen and blood urea nitrogen (DUN/BUN) and the expressions of CTGF and HSP47 were increased (all P＜0.05).After application of 1,25-dihydroxyvitamin D3,peritoneal fibrosis lesion was significantly improved,the peritoneum thickness,the expressions of CTGF and HSP47 were decreased (all P ＜ 0.05).In vitro,2.5％ high glucose induced-peritoneal mesothelial cells were respectively treated by CTGF siRNA,1,25-(OH)2-VitD3 and combined interventions,the expression of FN,CTGF and HSP47 was significantly lower than that in high glucose group (all P ＜ 0.05).Conclusions The expression of CTGF and HSP47 is significantly increased in peritoneal fibrosis rats.1,25-(OH)2-VitD3 may ameliorate the progression of peritoneal fibrosis via reducing the expression of CTGF,decreasing the expression of HSP47 and FN.

In-silico analysis of heat shock protein 47 for identifying the novel therapeutic agents in the management of oral submucous fibrosis.

Background: Heat shock proteins-47 (HSP47) is a collagen specific molecular chaperone, involved in the processing and/or secretion of procollagen. It seems to be regularly upregulated in various fibrotic or collagen disorders. Hence, this protein can be a potential target for the treatment of various fibrotic diseases including oral submucous fibrosis (OSF), which is a collagen metabolic disorder of oral cavity and whose etiopathogeneic mechanism and therapeutic protocols are still not well documented. Aim: The aim of this study is to identify the novel therapeutic agents using in-silico methods for the management of OSF. Objectives: The objectives of this study are to identify the binding sites of HSP47 on the collagen molecule and to identify the lead compound with anti-HSP47 activity from the library of natural compounds, using in-silico methodology. Materials and Methods: The web-based and tool based in-silico analysis of the HSP47 and collagen molecules are used in this study. The crystal structure of collagen and HSP47 were retrieved from Protein Data Bank website. The binding site identification and the docking studies are done using Molegro Virtual Docker offline tool. Results: Out of the 104 Natural compounds, six ligands are found to possess best binding affinity to the binding amino acid residues. Silymarin binds with the 4AU2A receptor and the energy value are found to be −178.193 with four Hbonds. The other best five natural compounds are hesperidin, ginkgolides, withanolides, resveratrol, and gingerol. Our findings provide the basis for the in-vitro validation of the above specified compounds, which can possibly act as "lead" molecules in designing the drugs for OSF. Conclusion: HSP47 can be a potential candidate to target, in order to control the production of abundance collagen in OSF. Hence, the binding sites of HSP47 with collagen are identified and some natural compounds with a potential to bind with these binding receptors are also recognized. These natural compounds might act as anti-HSP47 lead molecules in designing novel therapeutic agents for OSF, which are so far unavailable.

Dynamic expression of heat shock protein 47 in filtering bleb following the glaucomatous filtering surgery in rat / 中华实验眼科杂志

Background Anti-scarring is a focus following glaucoma filtering surgery.It has been verified that expression of heat shock protein 47 (HSP47) is always correlated with that of various types of collagens and closely related with the collagen-related diseases including fibrosis in various organs.However,the relationship between the expression of HSP47 and fibrosis of filtering bleb is unclear.Ohjective Present study was to investigate the dynamic expression of HSP47 in subconjunctival filtering bleb after anti-glaucomatous filtering surgery.Methods Filtering surgery was performed on the right eyes of 10 SPF male Spargue-Dawley(SD) rats.Then the operative eyes were randomized into the postoperative 7-day group and 14-day group according to the randomized number table,and the left eyes of the rats served as the normal control group.The shape of filtering blebs and the ocular anterior segment response were examined under microscope daily.The animals were sacrificed by excessive anesthesia on day 7 and 14 after operation,respectively,and the specimens were prepared of the eyes.Regular histopathological examination was performed to observe the collagen fibrosis change,and immunohistochemistry was carried out to assay the dynamic expression of HSP47 depended on the fibrosis.Percentage of positive cells for HSP47 were calculated.The use and care of the experimental animals complied with the Regulation for the Administration of Affair Concerning the Experimental Animals by State Science and Technology Committee.Results Diffuse subconjunctival blebs were seen postoperatively in t0 eyes after surgery with mild inflammatory response in ocular anterior segment.Hematoxylin-eosin staining showed that compared with the specimens of the normal control group,fibroblasts obviously proliferated and collagenous fibers increased in the specimens of operative group.The HSP47 protein was less expressed in cytoplasm of many fibroblasts and less vascular endothelial cells in the normal rat specimens.The expressing intensity of HSP47 protein was enhanced.However,the expression intensity was weaker in the postoperative 14-day group in comparison with the 7-day group.The percentage of positive cells for HSP47 in the filtering blebs 7 days after operation was (56.40±1.12)％,that in the normal control group was (13.70±0.74)％,with a significant difference between them (P=0.000).In the postoperative 14-day group,the percentage of positive cells for HSP47 reduced from (56.40± 1.12) ％ to (23.90±0.76) ％ (P =0.000),but it was still higher than that of the normal control group (P =0.000).Conclusions Expression intensity of HSP47 alters as the activity of fibroblasts in filtering bleb,suggesting that HSP47 plays a role in collagen-fibrosis following filtering surgery in SD rats.

Angiotensin Ⅱ induces the release of inflammation factors via TLR4-MyD88 pathway in rat tubular epithelial cells / 中华肾脏病杂志

Objective To observe the release of inflammation-related factors after angiotensin Ⅱ (Ang Ⅱ ) stimulation in rat tubular epithelial cells (NRK-52E), to analyze whether these effects were mediated by TLR4-MyD88 pathway, and to reveal the novel mechanism of injury by Ang Ⅱ on NRK-52E cells. Methods After synchronization, cells incubated with AngⅡ (10-7 mmol/L) were used as the stimulation group, cells without stimulation were as normal control. To determine the role of TLR4 and the adaptor MyD88, equal number of NRK-52E cells was added with 10-5 mmol/L candesartan or 20 mg/L TLR4 blocking peptide for 1 h and then incubated with Ang Ⅱ (10-7 mmol/L) respectively. RT-PCR was used to analyze TLR4 mRNA and MyD88 mRNA expression. Immunofluorescence and confocal microscopy were used to observe TLR4 protein expression. ELISA was used to detect the concentration of tumor necrosis factor-alpha (TNF-α) and heat shock protein 47(HSP47) in cell supernatant respectively. Results TLR4 and MyD88 were highly expressed in Ang Ⅱ-induced NRK-52E cells (P＜0.01), and the TNF-α and HSP47 levels were also increased markedly compared with control group (P＜0.01). In NRK-52E cells that were pre-incubated with candesartan, TLR4 and MyD88 expression were obviously inhibited,subsequently, HSP47 and TNF-α production decreased remarkably compared with Ang Ⅱ group (P＜0.01). TLR4 blocking peptide had the similar effect in a dose-dependent manner, in which its effect was dependent on inhibiting TLR4-MyD88 expression. Conclusion The mechanism of Ang Ⅱ -induced injury effect on NRK-52E cells is related to the increase of TLR4-MyD88 activity,which is followed by the enhance of TNF-α and HSP47 expression. This process is inhibited by candesartan via modulation of innate immune pathway.

Effects of RNAi on keloid formation by inhibiting heat shock protein / 中华医学美学美容杂志

Objective To study the significance of HSP47 gene in keloid formation after in vivo study with RNAi technology and the recombinant HSP4 7 siRNA against heat shock protein 47 to keloid in a nude mice model.Methods We injected RNAi mixture into the keloid of a nude mice model in experimental group and PBS water(0.25 ml)into control group at the 16th days after establishing the models.After interference we observed the specimens and harvested specimens at 7th days for biochemical and pathological analysis.Results The expression of HSP47 mRNA reduced obviously and the collagen content also reduced in the experimental group.The rusults had statistical significance.Conclusion We can suppress the expression of HSP47 gene and then reduce the production of collagen after in vivo interfering experiment with HSP4 7 siRNA in keloid nude mice models using RNAi technique.This study cornfirms the mechanism that HSP47 promotes the keloid formation,which provides a new target to treat keloid.