Protective effects of Baicalin injection on severe acute pancreatitis through regulating follistatin-like-1 signaling pathway by down-regulating miR-429 expression in mice

Abstract In China, Scutellaria is used for treating inflammatory-related diseases. Baicalin is the main active component of Scutellaria and has protective effects on acute pancreatitis. However, the mechanism of Baicalin is still unclear. In this study, the protective effects of baicalin on acute pancreatitis induced by taurocholate and its mechanism are investigated. In this study, mice were randomly divided into three groups: sham operation, model, and treatment groups. Acute pancreatitis in mice was induced by intraperitoneal injection of taurocholate (35 mg/kg). The treatment group was given baicalin (100 mg/kg) 2 h before acute pancreatitis induction. The mRNA expression levels of miR-429, nuclear factor kappa B65(NF-kB65), toll-like receptor 4(TLR4), TNF receptor associated factor6 (TRAF6), NF-kappa-B inhibitor(IkB), Follistatin-like 1 (FSTL1), and interleukin-1 receptor-associated kinase (IRAK) in the liver tissues 24 h after intraperitoneal injection were detected by RT-PCR. Then, the expression levels of NF-kB65, p-NF-κB65, TLR4, TRAF6, IkB, FSTL1, IRAK, p- IRAK, and p- IkB-а proteins were detected by Western blot. IL-6, TNF-α and IL-1 ß in plasma were measured by ELISA, and histopathological changes in the pancreases of the mice were observed. The results showed that after baicalin treatment, miR-429 expression in the pancreatic tissues and the expression levels of NF-kB65, TLR4, TRAF6, p-IkB-а, FSTL1, and p-IRAK decreased. Similarly, pancreatic myeloperoxidase (MPO) activity and the plasma levels of IL-6, TNF-а, IL-12, IL-1ß1, endotoxin, serum amylase, and lipase were reduced. Thus, the pancreatic injury induced by taurocholate was alleviated. The present study indicates that pretreatment with Baicalin can alleviate acute pancreatic injury induced by taurocholate in mice. The mechanism may be associated with the decreased miR-429 expression, reduced FSTL1 signaling pathway activity, TLR4 and TLR4/MyD88 signaling pathway inhibition, and reduced pancreatic inflammation. FSTL1 is the regulatory target for miR-429

Follistatin Mitigates Myofibroblast Differentiation and Collagen Synthesis of Fibroblasts from Scar Tissue around Injured Flexor Tendons

Follistatin-Like 1 Promotes Bleomycin-Induced Pulmonary Fibrosis through the Transforming Growth Factor Beta 1/Mitogen-Activated Protein Kinase Signaling Pathway / 中华医学杂志(英文版)

Background@#Follistatin-like 1 (FSTL1) is a novel profibrogenic factor that induces pulmonary fibrosis (PF) through the transforming growth factor-beta 1 (TGF-β1)/Smad signaling. Little is known about its effects on PF through the non-Smad signaling, like the mitogen-activated protein kinase (MAPK) pathway. Therefore, this study aimed to investigate the role of FSTL1 in PF through the MAPK signaling pathway and its mechanisms in lung fibrogenesis.@*Methods@#PF was induced in Fstl1and wild-type (WT) C57BL/6 mice with bleomycin. After 14 days, the mice were sacrificed, and lung tissues were stained with hematoxylin and eosin; the hydroxyproline content was measured to confirm PF. The mRNA and protein level of FSTL1 and the change of MAPK phosphorylation were measured by quantitative polymerase chain reaction and Western blotting. The effect of Fstl1 deficiency on fibroblasts differentiation was measured by Western blotting and cell immunofluorescence. MAPK signaling activation was measured by Western blotting in Fstl1 and WT fibroblasts treated with recombinant human FSTL1 protein. We pretreated mouse lung fibroblast cells with inhibitors of the extracellular signal-regulated kinase (ERK), p38, and Jun N-terminal kinase (JNK) signaling and analyzed their differentiation, proliferation, migration, and invasion by Western blotting, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide analysis, and transwell assays. The Student's t-test was used to compare the differences between two groups.@*Results@#Fstl1 deficiency attenuated phosphorylation of the ERK, p38, and JNK signaling in bleomycin-induced fibrotic lung tissue 14 days after injury (0.67 ± 0.05 vs. 1.22 ± 0.03, t = 14.92, P = 0.0001; 0.41 ± 0.01 vs. 1.15 ± 0.07; t = 11.19; P = 0.0004; and 0.41 ± 0.01 vs. 1.07 ± 0.07, t = 8.92, P = 0.0009; respectively), compared with WT lungs at the same time and in primary lung fibroblasts (0.82 ± 0.01 vs. 1.01 ± 0.04, t = 4.06, P = 0.0150; 1.04 ± 0.03 vs. 1.24 ± 0.03, t = 4.44, P = 0.0100; and 0.76 ± 0.05 vs. 0.99 ± 0.05, t = 4.48, P = 0.0100; respectively), compared with TGF-β1-stimulated WT group. Recombinant human FSTL1 protein in lung fibroblasts enhanced TGF-β1-mediated phosphorylation of the ERK (1.19 ± 0.08 vs. 0.55 ± 0.04, t = 6.99, P = 0.0020), p38 (1.18 ± 0.04 vs. 0.66 ± 0.03, t = 11.20, P = 0.0020), and JNK (1.11 ± 0.01 vs. 0.84 ± 0.04, t = 6.53, P = 0.0030), compared with the TGF-β1-stimulated WT group. Fstl1-deficient fibroblasts showed reduced alpha-smooth muscle actin (α-SMA) expression (0.70 ± 0.06 vs. 1.28 ± 0.11, t = 4.65, P = 0.0035, compared with the untreated WT group; 1.40 ± 0.05 vs. 1.76 ± 0.02, t = 6.31, P = 0.0007; compared with the TGF-β1-treated WT group). Compared with the corresponding condition in the control group, the TGF-β1/FSTL1-mediated α-SMA expression was significantly suppressed by pretreatment with an inhibitor of p38 (0.73 ± 0.01 vs. 1.13 ± 0.10, t = 3.92, P = 0.0078) and JNK (0.78 ± 0.03 vs. 1.08 ± 0.06, t = 4.40, P = 0.0046) signaling. The proliferation of mouse lung fibroblast cells (MLgs) significantly decreased after treatment of an inhibitor of p38 (0.30 ± 0.01 vs. 0.46 ± 0.03, t = 4.64, P = 0.0009), JNK (0.30 ± 0.01 vs. 0.49 ± 0.01, t = 12.84, P = 0.0001), and Smad2/3 (0.18 ± 0.02 vs. 0.46 ± 0.02, t = 12.69, P = 0.0001) signaling compared with the dimethylsulfoxide group. The migration and invasion cells of MLgs significantly decreased in medium pretreated with an inhibitor of p38 (70.17 ± 3.28 vs. 116.30 ± 7.11, t = 5.89, P = 0.0042 for the migratory cells; 19.87 ± 0.84 vs. 32.70 ± 0.95, t = 10.14, P = 0.0005 for the invasive cells), JNK (72.30 ± 3.85 vs. 116.30 ± 7.11, t = 5.44, P = 0.0056 for the migratory cells; 18.03 ± 0.94 vs. 32.70 ± 0.95, t = 11.00, P = 0.0004 for the invasive cells), and Smad2/3 (64.76 ± 1.41 vs. 116.30 ± 7.11, t = 7.11, P = 0.0021 for the migratory cells; 18.03 ± 0.94 vs. 32.70 ± 0.95, t = 13.29, P = 0.0002 for the invasive cells) signaling compared with the corresponding condition in the dimethylsulfoxide group.@*Conclusion@#FSTL1 affects lung fibroblast differentiation, proliferation, migration, and invasion through p38 and JNK signaling, and in this way, it might influence the development of PF.

Follistatin-like protein 1 plays a tumor suppressor role in clear-cell renal cell carcinoma / 癌症

BACKGROUND@#We previously showed that the expression of follistatin-like protein 1 (FSTL1) was significantly down-regulated in metastatic clear-cell renal cell carcinoma (ccRCC). In this study, we aimed to characterize the role of FSTL1 in the development of ccRCC.@*METHODS@#The effects of FSTL1 on cell activity and cell cycle were investigated in ccRCC cell lines with altered FSTL1 expression. Gene expression microarray assays were performed to identify the major signaling pathways affected by FSTL1 knockdown. The expression of FSTL1 in ccRCC and its effect on postoperative prognosis were estimated in a cohort with 89 patients.@*RESULTS@#FSTL1 knockdown promoted anchorage-independent growth, migration, invasion, and cell cycle of ccRCC cell lines, whereas FSTL1 overexpression attenuated cell migration. FSTL1 knockdown up-regulated nuclear factor-κB (NF-κB) and hypoxia-inducible factor (HIF) signaling pathways, increased epithelial-to-mesenchymal transition, up-regulated interleukin-6 expression, and promoted tumor necrosis factor-α-induced degradation of NF-κB inhibitor (IκBα) in ccRCC cell lines. FSTL1 immunostaining was selectively positive in epithelial cytoplasm in the loop of Henle, and positive rate of FSTL1 was significantly lower in ccRCC tissues than in adjacent renal tissues (P < 0.001). The multivariate Cox regression analysis showed that the intratumoral FSTL1 expression conferred a favorable independent prognosis with a hazard ratio of 0.325 (95% confidence interval 0.118-0.894). HIF-2α expression was negatively correlated with FSTL1 expression in ccRCC specimens (r = - 0.229, P = 0.044). Intratumoral expression of HIF-2α, rather than HIF-1α, significantly predicted an unfavorable prognosis in ccRCC (log-rank, P = 0.038).@*CONCLUSIONS@#FSTL1 plays a tumor suppression role possibly via repressing the NF-κB and HIF-2α signaling pathways. To increase FSTL1 expression might be a candidate therapeutic strategy for metastatic ccRCC.

Expression level and clinical significance of FSTL1 in serum of patients with acute coronary syndrome / 国际检验医学杂志

Objective To observe the level changes and clinical diagnostic value of follicular statin -1 (FSTL1)in the serum of patients with different types of acute coronary syndrome(ACS).Methods Collected the clinical diagnosis of acute coronary syndrome patients 98 cases,which contained ST segment elevation my-ocardial infarction(STEMI)in 34 cases,non ST elevation myocardial infarction(NSTEMI)in 28 cases,unsta-ble angina pectoris(UA)in 36 cases,while the examination resuLts of healthy people as a control group of 20 cases.The Venous blood was collected and the FSTL1 levels of the 4 groups were detected by ELISA.Results The levels of Serum FSTL1 in ACS group was significantly higher than that in normal control group(P<0.05).Serum FSTL1 of the ACS group were significant correlated with Gensini score,cTNT,hs-CRP(related coefficient:0.210,0.236,0.219 separately).The AUC of FSTL1 was 0.910(95% CI:0.832 -0.988),which was lower than cTNT.The best cut-off value of FSTL1 as a biomarker was 5.65 μg/L(specificity:84.2% and sensitivity:77.5%).Moreover the combination of FSTL1,HDL and cTNT exhibited significantly higher AUC=0.945(95% CI:0.909 -0.981)than did other biomarkers alone or pair combinations.Conclusion In pa-tients with acute coronary syndrome,serum FSTL1 levels has a positive correlation with the degree of coro-nary stenosis and inflammation reaction,and has certain value in the diagnosis of acute coronary syndrome.

Deficiency of Follistatin-Like Protein 1 Accelerates the Growth of Breast Cancer Cells at Lung Metastatic Sites / 한국유방암학회지

PURPOSE: Follistatin-like protein 1 (FSTL1) is a secreted glycoprotein that has been shown to play a role in various types of cancer. However, the clinical significance and function of FSTL1 in breast cancer have not been reported. We investigated the role of FSTL1 in breast cancer in this study. METHODS: Enzyme-linked immunosorbent assays, western blot analysis, and reverse transcription polymerase chain reaction were used to monitor the expression of FSTL1 in breast cancer tissue and in serum samples from breast cancer patients. We employed a 4T1 breast cancer model and Fstl1(+/−) mice for in vivo studies. Hematoxylin and eosin staining, western blot analysis, and RNA sequencing were used to analyze the effect of FSTL1 on primary tumor growth and lung metastasis. RESULTS: We demonstrated that the expression of FSTL1 is reduced in both the breast cancer tissue and the serum of breast cancer patients. We showed that reduced levels of FSTL1 in serum correlate with elevated expression of Ki-67 and epidermal growth factor receptor (EGFR) in cancer tissues. Moreover, lowered expression of FSTL1 was associated with decreased survival in breast cancer patients. Experiments on the Fstl1(+/−) mouse model established that FSTL1 deficiency had no effect on primary tumor growth, but increased the lung metastases of breast cancer cells, resulting in reduced survival of tumor-bearing mice. RNA sequencing found significantly reduced expression of Egln3 and increased expression of EGFR in Fstl1(+/−) mice. Thus, our results suggest that FSTL1 may affect the expression of EGFR through Egln3, inhibiting the proliferation of breast cancer cells at lung metastatic sites. CONCLUSION: In conclusion, we suggest a suppressor role of FSTL1 in breast cancer lung metastasis. Furthermore, FSTL1 may represent a potential prognostic biomarker and a candidate therapeutic target in breast cancer patients.

Effect of follistatin on skeletal muscle wasting of cancer cachexia mice and its mechanism / 吉林大学学报(医学版)

Objective: To observe the effects of follistatin (FST)on the skeletal muscle wasting of cancer cachexia mice and the expressions of Mstn, LncRNA-MALAT1 and Caspase-3, and to elucidate its associated molecular mechanisms.Methods:Thirty-two BALB/c mices were randomly assigned into:healthy control (HC) group,FST prevention (FP)group,FST treatment (FT)group and cancer cachexia (CC)group.The murine colon adenocarcinoma CT26 cells were inoculated subcutaneously into the mices in FP, FT and CC groups to establish the cancer cachexia models. The body weight, spontaneous activity and tumor growth were daily monitored.The mice in FP and FT groups were administrated with FST intraperitoneally on day 6 and 12 after inoculation.The samples were collected on day 20.The tumor and gastrocnemius weights of the mice were detected. The biochemical metabolism indexes and myofiber cross-sectional area of gastrocnemius tissue were detected.The mRNA expression levels of Mstn,Caspase-3 and LncRNA-MALAT1 were examined by Real-time PCR.The protein expression levels of Mstn and Caspase-3 were measured by Western blotting method. Results:Compared with CC group,the body weights,spontaneous activities,gastrocnemius weights and myofiber cross-sectional areas were increased (P <0.05);the serum levels of glucose,total protein and albumin of the mice in FP and FT groups were increased (P <0.05).The protein and mRNA expression levels of Mstn and Caspase-3 in gastrocnemius of the mice in CC group were significantly higher and the expression level of LncRNA-MALAT1 was significantly lower than those in HC group (P < 0.05).The mRNA and protein expression levels of Mstn and Caspase-3 in FP and FT groups were reduced and the expression level of LncRNA-MALAT1 was increased compared with CC group (P < 0.05).The prevention effect in FP group is better than FT group (P < 0.05). Conclusion:FST may alleviate the muscle wasting of the mice with cancer cachexia by inhibiting the expression of Mstn,thus upregulating the expression of LncRNA-MALAT1 which in turn to suppress the expression of Caspase-3.

Effects of cobalt chloride-induced hypoxia on cell migration and expression and secretion of FSTL1 in melanoma cell line / 天津医药

Objective To explore the effects of cobalt chloride (CoCl2)-induced hypoxia on migration of melanoma cells, and to detect the transcription, expression and secretion of Follistatin-like 1(FSTL1) in this process. Methods B16F10 melanoma cell line was treated with CoCl2 in order to mimic hypoxia. Experimental cells were divided into three groups: 0μmol/L, 50μmol/L and 100μmol/L CoCl2 treatment groups. MTT assay was used to assure cell viability, and to determine the treatment concentration of CoCl2. Transwell assay was used to determine the migration ability of B16F10 melanoma cell line. Real-time PCR was used to measure the mRNA expression of Fstl1. Western blot assay was used to detect the intracel?lular and extracellular protein expression of FSTL1. Results The cell viability of B16F10 melanoma cell line was signifi?cantly reduced by CoCl2 treatment, with a time and concentration-dependent manner. The migration ability of B16F10 cell line was significantly increased in CoCl2 treated group compared with that of control group (P<0.05). The mRNA level of Fstl1 was obviously higher in CoCl2 treated group than that of control group (P<0.05). The intracellular expression of FSTL1 protein was consistent with the expression trend of Fstl1 mRNA. Simultaneously, the extracellular protein level of FSTL1 was significantly decreased compared with that of control group. There was no expression of FSTL1 in 100μmol/L CoCl2 treat?ment group. Conclusion The migration ability of melanoma cell line is enhanced by CoCl2 treatment, which may be associ?ated with expression and secretion of FSTL1, however, the relevant mechanism still needs further investigation.

Effect of physical training on liver expression of activin A and follistatin in a nonalcoholic fatty liver disease model in rats

Nonalcoholic fatty liver disease (NAFLD) is characterized by fat accumulation in the liver and is associated with obesity and insulin resistance. Activin A is a member of the transforming growth factor beta (TGF)-β superfamily and inhibits hepatocyte growth. Follistatin antagonizes the biological actions of activin. Exercise is an important therapeutic strategy to reduce the metabolic effects of obesity. We evaluated the pattern of activin A and follistatin liver expression in obese rats subjected to swimming exercise. Control rats (C) and high-fat (HF) diet-fed rats were randomly assigned to a swimming training group (C-Swim and HF-Swim) or a sedentary group (C-Sed and HF-Sed). Activin βA subunit mRNA expression was significantly higher in HF-Swim than in HF-Sed rats. Follistatin mRNA expression was significantly lower in C-Swim and HF-Swim than in either C-Sed or HF-Sed animals. There was no evidence of steatosis or inflammation in C rats. In contrast, in HF animals the severity of steatosis ranged from grade 1 to grade 3. The extent of liver parenchyma damage was less in HF-Swim animals, with the severity of steatosis ranging from grade 0 to grade 1. These data showed that exercise may reduce the deleterious effects of a high-fat diet on the liver, suggesting that the local expression of activin-follistatin may be involved.

Construction of a eukaryotic expression vector for pEGFP-FST and its biological activity in duck myoblasts

Background Follistatin (FST), a secreted glycoprotein, is intrinsically linked to muscle hypertrophy. To explore the function of duck FST in myoblast proliferation and differentiation, the pEGFP-FST eukaryotic expression vector was constructed and identified. The biological activities of this vector were analyzed by transfecting pEGFP-FST into cultured duck myoblasts using Lipofectamine™ 2000 and subsequently determining the mRNA expression profiles of FST and myostatin (MSTN). Results The duck pEGFP-FST vector was successfully constructed and was confirmed to have high liposome-mediated transfection efficiency in duck myoblasts. Additionally, myoblasts transfected with pEGFP-FST had a higher biological activity. Significantly, the overexpression of FST in these cells significantly inhibited the mRNA expression of MSTN (a target gene that is negatively regulated by FST). Conclusions The duck pEGFP-FST vector has been constructed successfully and exhibits biological activity by promoting myoblast proliferation and differentiation in vitro.

Uso tópico de 5-azacitidina melhora a cicatrização de feridas cutâneas de roedores por meio do sistema ativina/folistatina / Topical 5-azacytidine accelerates skin wound healing in rats

O desenvolvimento de métodos que tem por objetivo acelerar e melhorar a qualidade do processo de cicatrização de feridas tem impacto positivo na condução de distúrbios de cicatrização associados a inúmeras condições médicas. Neste estudo, avaliamos os efeitos moleculares, celulares e clínicos da aplicação tópica de 5-azacitidina na cicatrização de feridas em ratos. De acordo com estudos pregressos, a 5-azacitidina reduz a expressão de folistatina, que é um regulador negativo das ativinas. Estas, por sua vez, promovem o crescimento de células em diferentes tecidos, incluindo a pele. Ratos Wistar machos com oito semanas de vida foram submetidos a um ferimento cutâneo com punch de oito milímetros na região dorsal. A seguir os ratos foram aleatoriamente separados em grupo controle (veículo) ou submetidos à aplicação tópica de 5-azacitidina (10 mM), uma vez por dia por até 12 dias, iniciando-se no terceiro dia após a lesão. A documentação fotográfica e coleta de amostras ocorreram nos dias 5, 9 e 15. O emprego desta droga resultou em aceleração da cicatrização da ferida, (99,7±7,0% versus 71,2±2,8% no dia 15, p <0,01). Este resultado clínico foi acompanhado pela redução de aproximadamente três vezes na expressão protéica de folistatina. O exame histológico da pele revelou re-epitelização eficiente com aumento da expressão de queratinócitos e aumento significativo na expressão do gene de TGF-β além da diminuição significativa de citocinas, tais como TNF-α e IL-10. Analisamos também a proliferação celular na lesão de pele através do método de incorporação de BrdU. O número de células positivas para BrdU aumentou significativamente quando comparado ao controle. No entanto, quando folistatina exógena foi aplicada na pele em paralelo ao tratamento tópico de 5-azacitidina a maioria dos benefícios do medicamento foi perdida.

The development of new methods aimed at improving wound healing may have an impact on the outcomes of a number of medical conditions. Here we evaluate the molecular and clinical effects of topical 5-azacytidine, a compound used in myelodysplasia, on the wound healing in rats. According to previous studies, 5-Azacytidine decreases the expression of follistatin 1, which is a negative regulator of activins. Activins, in turn, promote cell growth in different tissues, including the skin. Eight-week old male Wistar rats were submitted to an 8 mm punchwound in the dorsal region. After three days, rats were randomly assigned to either control or topical application of a solution containing 5-azacytidine (10mM), once a day. Photo documentation and collection of samples occurred at days 5, 9 and 15. Overall, 5-azacytidine resulted on a significant acceleration of complete wound healing (99.7% ±0.7.0 vs. 71.2%±2.8 on days 15; n=10; p<0,01). This was accompanied by an up to 3-fold reduction in follistatin expression. Histological examination of the skin revealed efficient reepithelization with increase in gene expression of TGF-β and keratinocytes markers, involucrin and citokeratin, besides the significant decrease of cytokines such as TNF-α and IL-10. In addition, we analyzed cell proliferation in injured skin employing the BrdU incorporation method. The treatment with 5-azacytidine led to a progressive increase of BrdU positive cells. Finally when recombinant follistatin was employed in the skin in parallel to topical 5-azacytidine most of the benefits of the drug were lost. Thus, 5-azacytidine acts, at least in part, through the follistatin/activin pathway to improve wound healing in rats. This study belongs to online research process Caring in Nursing and Health.

Expression of follistatin-like protein 1 in the peripheral blood and intestinal tissue of patients with ulcerative colitis and its clinical significance / 中华消化杂志

Objective To examine the expression of follistatin-like protein 1 (FSTL1) in the peripheral blood and intestinal mucosa tissues of ulcerative colitis (UC) patients and analyze the correlation between its expression and the activity of UC.Methods From October 2010 to June 2012,sixty patients with UC were collected.From April 2012 to October 2012,thirty individuals without any obvious mucosa lesion under colonoscope and confirmed by pathological examination were set as control group.The serum expression level of FSTL1 of both UC group and control group were determined by enzyme linked immunosorbent assay (ELISA).t-test was performed for comparison between groups.The expression of FSTL1 in the intestinal mucosa of UC group and control group was detected by immunohistochemistry.Chi-square test was used for comparison between groups.The patients with UC were scored with ulcerative colitis disease activity index (UCDAI).Its correlation with plasma FSTL1 was analyzed by Pearson correlation coefficient.Results The serum expression level of FSTL1 of UC group ((14.37-±-1.80) μg/L) was higher than that of control group ((5.80±0.72) μg/L)and the difference was statistically significant (t=25.01,P＜ 0.05).The serum expression level of FSTL1 of UC group was positively correlated with UCDAI (r=0.814,P＜0.05).The positive expression rate of FSTL1 in the intestinal mucosa tissues of UC group (86.7％,52/60)was higher than that of control group (46.7％,14/30) and the difference was statistically significant (x2 =52.334,P＜0.05).Conclusions The expression of FSTL1 of UC patients increases and is positively correlated with disease activity.FSTL1 may play a role in the development of UC.

The expression and clinical significance of serum follistatin-like protein 1 in patients with systemic lupus erythematosus / 中华风湿病学杂志

ObjectiveTo detect the serum level of follistatin-like protein 1 (FSTL1) in patients with systemic lupus erythematosus (SLE) and its expression in renal biopsy tissues in lupus nephritis (LN) patients as well as its clinical significance were analyzed.MethodsThe serum concentration of FSTL1 in 54 SLE patients and 27 healthy controls was measured with enzyme-linked immunosorbent assay (ELISA).The distribution of FSTL1 in renal biopsy tissues was stained by immune-histochemical method.Mann-Whitney U test,t test,X2test and Pearson test were selected to compare the changes and data analysis.ResultsThe serum FSTLI level was significantly higher in SLE patients(26±21) μg/L than those of healthy controls ( 12± 14) μg/L (P＜0.01).The level of serum FSTL1 was significantly higher in SLE patients with hypertension than in patients without hypertension.The serum FSTL1 level had statistically significant changes between SLE patients with disease duration ≥ 5 years and ＜5 years.The level of serum FSTL1 correlated positively with SLEDAI score (r=0.319,P=0.022),age (r=0.700,P＜0.01),disease duration (r=0.513,P＜0.01),complement C4 level (r=0.443,P=0.004),and total serum cholesterol level (r=0.460,P=0.001 ).FSTL1 correlated inversely with platelet count (r=-0.422,P =0.001 ),anti-dsDNA antibody levels (r=-0.276,P=0.046).FSTL1 expression was evident in the cytoplasm of epithelial cells of kidney tubules.ConclusionThe level of serum FSTL1 is significantly increased in SLE patients.FSTL1 concentration correlats positively with disease activity.These data indicate that FSTL1 may play a role in the pathogenesis of SLE.

Expressions of BMP Antagonists in Shh -/- ; Gli3 -/- Mutant / 체질인류학회지

Sonic hedgehog (Shh) has been known as an essential morphogen for the generation of motor neuron in developing spinal cord. However, motor neuron can be generated in Shh -/- ; Gli3 -/- or Gli2 -/- ; Gli3 -/- mutants although these mutants don't have Shh signaling. To find out the compensatory mechanism for the generation of motor neuron in Shh -/- ; Gli3 -/- mutant, we studied bone morphogenetic protein (BMP) antagonists including follistatin, flik and noggin, and retinoic acid signaling in this mutant. To study expressions of BMP antagonists, we performed in situ hybridization. To examine an activity of retinoic acid, we measured beta -galactosidase activity in retinoic acid response element (RARE) transgenic mouse. The expression of follistatin was reduced at both levels of forelimb and hindlimb in Shh -/- mutant compared to wild type embryo. It was restored at the level of forelimb but reduced at the level of hindlimb in Shh -/- ; Gli3 -/- mutant compared to wild type. The expression of flik was similar with wild type embryo at both levels of forelimb and hindlimb in Shh -/- mutant. The expression of flik was similar with wild type embryo at the level of forelimb however reduced in hindlimb level in Shh -/- ; Gli3 -/- mutant. The expression of noggin, a BMP antagonist, was increased in Shh -/- mutant. Activity of retinoic acid signaling was not affected in Shh -/- or Shh -/- ; Gli3 -/- mutants. From these results, we conclude that retinoic acid but not follistatin and flik, may be involved in the generation of motor neuron in Shh -/- ; Gli3 -/- mutant.

Cloning and Expression Pattern of FSTL1 mRNA in Normal Myometrium and Uterine Leiomyoma / 대한산부인과학회잡지

OBJECTIVE: To study the influence and cloning of differentially expressed genes in human female normal myometrium and uterine leiomyoma tissue. METHODS: In this experiment, human uterus tissues (n=25) were taken for total RNA isolation by using Trizol reagent. Differential display was performed by using GeneFishingTM DEG Kit and processed to cDNA sequencing and gene cloning for Follistatin-like 1 (FSTL1). Data were analyzed with the image Master VDS software and statistical significance was defined as p<0.05 by paired t test results. RESULTS: FSTL1 mRNA expression level was significantly higher (p<0.05) in normal and adjacent normal myometrium tissues than uterine leiomyoma tissue of women in the reproductive age. Whereas in the menopausal age, FSTL1 mRNA expression level was significantly higher (p<0.05) in uterine leiomyoma than normal myometrium. There was no significant differences between uterine leiomyoma and adjacent normal myometrium. CONCLUSION: Although the mechanisms of FSTL1 gene were uncertain, FSTL1 seemed to play an important role in the growth of uterine leiomyoma, it also might be related to the regulation of uterine leiomyoma growth inhibiting factors by modulating Follistatin related protein gene (FLRG) system.

Influence of Activin and Follistatin on Expression of Insulin-like Growth Factor (IGF)-I, II, Insulin-like Growth Factor Binding Protein (IGFBP)-1, 2, and 3 mRNA in Cultured Mouse Granulosa Cells / 대한산부인과학회잡지

OBJECTIVE: To investigate the influence of activin and follistatin on the expression of IGF (insulin-like growth factor)-I, II, IGFBP (insulin-like growth factor binding protein)-1, 2, and 3 mRNA in cultured mouse granulosa cells MATERIALS AND METHODS: The granulosa cells were obtained from the mouse and cultured for 6 days with 10 ng/ml of activin, 10 ng/ml of follistatin, and 10 ng/ml of activin with 10 ng/m of follistatin, respectively. The cells not treated with activin or follistatin served as control. Reverse transcription-polymerase chain reaction (RT-PCR) has been used to examine the expression of IGF-I, II, IGFBP-1, 2, and 3 mRNA. Results were analyzed with Kolmogorov-Smirnov test and analysis of variance (ANOVA) and statistical significance was defined as p<0.05. RESULTS: The expression of IGF-I and II mRNA were not different significantly. However, the expression of IGFBP-3 mRNA was significantly increased in the follistatin group compared to the control group (p<0.05) and significantly decreased in the activin with follistatin group compared to the control group (p<0.05). The expression of IGFBP-1 mRNA was seemed to be increased in the activin group and decreased in the follistatin group compared to the control group, respectively (p=0.07, p=0.07). The expression of IGFBP-2 and 3 mRNA were seemed to be decreased in the activin group compared to the control group (p=0.06, p=0.07, respectively). CONCLUSION: Activin and follistatin might play a role as regulators of mouse ovarian physiology by modulating the IGF system, especially IGFBPs.

Effects of activin A and follistatin on type Ⅰ collagen of cultured rat renal interstitial fibroblasts / 解放军医学杂志

Objective To investigate the stimulating effects of activin A(ACT-A) and follistatin(FS) on the secretion of type I collagen(Col I) and the expression of Col I mRNA of cultured rat renal interstitial fibroblasts in vitro.Methods Rat renal interstitial fibroblasts isolated from normal SD rat renal medulla were cultured in vitro.The cells were divided into three groups:different concentrations of ACT-A(group A),different concentrations of FS(group F),and different concentrations of FS plus 30ng/ml ACT-A(group A+F).The expression of Col I mRNA in cultured rat renal interstitial fibroblasts was detected by RT-PCR,and the expression of Col I protein in cultured rat renal interstitial fibroblasts was examined by immunocytochemistry.Results Renal interstitial fibroblasts were successfully cultured in vitro.In present study,the expression of Col I mRNA increased significantly in cultured rat renal interstitial fibroblasts after stimulated by ACT-A in dose-dependent manner(P0.05).FS could inhibit the effects of ACT-A(30ng/ml) on Col I mRNA and Col I protein of the cultured renal interstitial fibroblasts in dose-dependent manner in A+F group(P

Effects of activin and follistatin on hydroxyproline and type Ⅳ collagen secreted by cultured rat renal interstitial fibroblasts / 第三军医大学学报

Objective To investigate the roles of activin A(ACT-A) and follistatin(FS) in renal interstitial fibrosis.Methods Renal interstitial fibroblasts were isolated from SD rat kidney and primarily cultured.The cells were divided into A group,F group and A+F group,and then each group was further divided into 5 subgroups according to their culture media being added with ACT-A(0,0.3,3,30,or 100 ng/ml),FS(0,0.3,3,30,or 100 ng/ml),and ACT-A(30 ng/ml) plus FS(0,0.3,3,30,or 100 ng/ml) respectively.Levels of type Ⅳ collagen(Ⅳ-C) and hydroxyproline(HYP) in the supernatants of cultured fibroblasts were measured by ELISA assay.Results Concentrations of Ⅳ-C and HYP in cultured rat renal interstitial fibroblasts were in a dose-dependent manner with ACT-A treatment.FS had no effect on these concentrations,but it inhibited the effects of ACT-A on Ⅳ-C and HYP in the supernatants of cultured fibroblasts in a dose-dependent manner.Conclusion ACT-A might be involved in occurrence and development of renal fibrosis by affecting renal interstitial fibroblasts.Exogenous FS could suppress the effects of ACT-A on the cultured renal fibroblasts.