anti human connective tissue growth factor  (R&D Systems)

Journal: Frontiers in Pharmacology

Article Title: Kinsenoside Protects Against Radiation-Induced Liver Fibrosis via Downregulating Connective Tissue Growth Factor Through TGF- β 1 Signaling

doi: 10.3389/fphar.2022.808576

Figure Lengend Snippet: The key fibrosis-related gene and signaling pathway that are inhibited by KD (A) The results of high-throughput sequencing of the transcriptome showed that the expression of CTGF gene increased after irradiation (B) The results of high-throughput sequencing of the transcriptome showed that the expression of CTGF gene decreased after the administration of KD (C) The results of Real-Time PCR were consistent with the results of the sequencing. The expression of CTGF mRNA increased in the IR group, but decreased in the IR + KD group (D) The results of western blotting were consistent with the results of sequencing. The expression of CTGF protein increased in the IR group, but decreased in the IR + KD group (E) In the transcriptome KEGG gene pathway enrichment map, the TGF-β pathway was closely related to CTGF and RILF (F) The results of tissue TGF-β1 immunohistochemistry indicated that in the IR + KD group the expression of TGF-β1 in the liver tissue decreased compared with the IR group. For all results in this figure, original magnification, ×40 and ×100. Mean ± SEM. * p < 0.05, ** p < 0.01.

Article Snippet: The antibodies included: α -SMA (1:1,000; Servicebio Technology, Wuhan, China), fibronectin (FN), collagen I and connective tissue growth factor (CTGF) (1:1,000; BOSTER Biological Technology, Wuhan, China), TGF- β 1 and Smad2/3 and P-Smad2/3 (1:1,000; Cell Signaling Technology Inc., Danvers, MA), GAPDH and β -actin (1:1,000; Aspen Biological, Wuhan, China).

Techniques: Next-Generation Sequencing, Expressing, Irradiation, Real-time Polymerase Chain Reaction, Sequencing, Western Blot, Immunohistochemistry

Journal: Frontiers in Pharmacology

Article Title: Kinsenoside Protects Against Radiation-Induced Liver Fibrosis via Downregulating Connective Tissue Growth Factor Through TGF- β 1 Signaling

doi: 10.3389/fphar.2022.808576

Figure Lengend Snippet: KD ameliorates the RILF by inhibiting TGF-β1/Smad/CTGF pathway (A) Exogenous TGF-β1 led to the increased expression of CTGF protein (B) Exogenous TGF-β1 induced an increase in the expression of collagen and a -SMA proteins (C) The TGF-β1 activity in the conditioned media significantly increased after 6Gy irradiation compared with the control group, while the TGF-β1 activity in the conditioned media in the IR + KD group decreased compared with the IR group (D-G) The expressions of TGF-β1 and downstream Smad2/3 as well as phosphorylated Smad2/3 reduced in the IR + KD group compared with the IR group. Mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: The antibodies included: α -SMA (1:1,000; Servicebio Technology, Wuhan, China), fibronectin (FN), collagen I and connective tissue growth factor (CTGF) (1:1,000; BOSTER Biological Technology, Wuhan, China), TGF- β 1 and Smad2/3 and P-Smad2/3 (1:1,000; Cell Signaling Technology Inc., Danvers, MA), GAPDH and β -actin (1:1,000; Aspen Biological, Wuhan, China).

Techniques: Expressing, Activity Assay, Irradiation, Control

Journal: Frontiers in Pharmacology

Article Title: Kinsenoside Protects Against Radiation-Induced Liver Fibrosis via Downregulating Connective Tissue Growth Factor Through TGF- β 1 Signaling

doi: 10.3389/fphar.2022.808576

Figure Lengend Snippet: The schematic diagram. KD ameliorated the RILF through the regulation of the key molecule CTGF and TGF-β1/Smad/CTGF pathway in the HSCs.

Article Snippet: The antibodies included: α -SMA (1:1,000; Servicebio Technology, Wuhan, China), fibronectin (FN), collagen I and connective tissue growth factor (CTGF) (1:1,000; BOSTER Biological Technology, Wuhan, China), TGF- β 1 and Smad2/3 and P-Smad2/3 (1:1,000; Cell Signaling Technology Inc., Danvers, MA), GAPDH and β -actin (1:1,000; Aspen Biological, Wuhan, China).

Techniques:

Journal: Journal of the American Society of Nephrology : JASN

Article Title: The Role of the EGF Receptor in Sex Differences in Kidney Injury

doi: 10.1681/ASN.2018121244

Figure Lengend Snippet: Male Dsk5 mice developed spontaneous kidney injury. Male wild-type and Dsk5 mice were euthanized at 15 weeks of age. (A) Male Dsk5 mice had kidney hypertrophy as indicated by increased kidney weight versus body weight (KW/BW) ratios (**P<0.01; n=7). (B) Male Dsk5 mice had increased albuminuria as indicated by increased urinary albumin-to-creatinine ratio (ACR) at 15 weeks of age (**P<0.01; n=10 in wild-type group and n=9 in Dsk5 group). (C) Period acid–Schiff staining showed that kidneys of male Dsk5 mice exhibited segmental and global glomerular sclerosis, mesangial expansion, hyaline, and protein casts. Original magnification, ×400 (scale bar, 25 µm). (D) Male Dsk5 mice had loss of podocytes as indicated by two representative photomicrographs from wild-type and Dsk5 mice (left panel) and quantitatively decreased WT1-positive cells (right panel; **P<0.01; n=3). Original magnification, ×400 (scale bar, 25 µm). (E) Male Dsk5 mice had epithelial cell injury as indicated by increased renal expression levels of KIM-1, a marker of proximal tubule epithelial injury. Original magnification, ×250 (scale bar, 40 µm). (F) Male Dsk5 mice exhibited tubulointerstitial fibrosis as indicated by Sirius Red and Masson trichrome staining. Original magnification, ×160 (scale bar, 62.5 µm). (G) Immunostaining and immunoblotting determined that male Dsk5 mice had increased renal expression levels of α-SMA, a marker of myofibroblasts, CTGF, and collagen I. Original magnification, ×160 for α-SMA (scale bar, 62.5 µm) and ×400 for collagen I (scale bar, 25 µm). Arrows indicate blood vessels. (H) Kidneys of male Dsk5 mice had increased mRNA levels of profibrotic and fibrotic components, including α-SMA, CTGF, TGF-β, collagen I and III (Col I and Col III), and fibronectin. *P<0.05; **P<0.01. n=7 in each group.

Article Snippet: Rat anti-mouse F4/80 (MCA497R, a marker of macrophages), CD68 (MCA1957), CD3 (MCA1477, a marker of T lymphocytes), CD4 (MCA2961), CD8a (MCA2694), and Ly-6G (Gr-1, MCA2387, a marker of neutrophils) were purchased from AbD Serotec (now Bio-Rad); goat anti–phospho-EGFR (Tyr 1173, SC-12351) and anti-human connective tissue growth factor (CTGF; SC-14939), rabbit anti–phospho-EGFR (Tyr845, SC-23420-R) and anti–IL-6 (SC-12912T) were purchased from Santa Cruz Biotechnology; rabbit anti-Wilms Tumor Protein (WT1, a marker of podocytes; ab89901) and TNF- α (ab6671) were from Abcam; goat anti-EGFR (E1282) and mouse anti– α -smooth muscle actin ( α -SMA; a marker of myofibroblasts, A5228) were from Sigma (St. Louis, MO); rabbit anti-collagen I (600–401–103–01) was purchased from Rockland Immunochemicals; rat anti–KIM-1 (a marker of kidney epithelial injury, MAB1817–100) and mouse anti–4-hydroxynonenal (a marker of oxidative stress, no. 198960) were from R&D Systems; and rabbit anti–p-ERK (4370S) was from Cell Signaling Technology.

Techniques: Staining, Expressing, Marker, Immunostaining, Western Blot

Journal: Journal of the American Society of Nephrology : JASN

Article Title: The Role of the EGF Receptor in Sex Differences in Kidney Injury

doi: 10.1681/ASN.2018121244

Figure Lengend Snippet: Castration led to decreases in renal EGFR expression and injury in male Dsk5 mice. (A) Male Dsk5 mice were castrated at 21 days of age and euthanized at 6 months of age. (B) Castration led to decreases in renal EGFR activation as indicated by phospho-EGFR (Y845) immunofluorescence staining, total renal EGFR expression, and p-ERK levels, a downstream signaling of EGFR activation. Original magnification, ×400 (scale bar, 20 µm). (C) Glomerular sclerosis, tubular dilation, and protein casts were apparent in sham male Dsk5 mice, whereas only mesangial expansion was seen in castrated male Dsk5 mice. Original magnification, ×160 (scale bar, 62.5 µm). (D) Loss of WT1 staining, a marker of podocytes, seen in male Dsk5 mice was significantly attenuated but was not completely prevented by castration (**P<0.01; ***P<0.001 versus wild-type group; ††P<0.01 versus sham Dsk5 group; n=6 in each group). Original magnification, ×400 (scale bar, 25 µm). (E) Tubular injury seen in male Dsk5 mice was dramatically attenuated by castration as indicated by significant reduction of renal expression levels of KIM-1 and NGAL, two markers of tubular injury (***P<0.001 versus wild-type group; †††P<0.001 versus sham Dsk5 group; n=8 in wild-type group and n=7 in other groups). (F) Castration prevented progressive albuminuria seen in male Dsk5 mice (*P<0.05; **P<0.01; ***P<0.001 versus sham Dsk5 mice; n=8 in wild-type group and n=7 in other groups). (G) Castration significantly reduced tubulointerstitial fibrosis in male Dsk5 mice as indicated by Sirius Red staining (***P<0.001 versus wild-type group; †††P<0.001 versus sham Dsk5 group; n=6). Original magnification, ×160 (scale bar, 80 µm). (H) Castration decreased renal expression levels of α-SMA and CTGF in male Dsk5 mice. (I) Male Dsk5 mice had significant increases in renal expression levels of HB-EGF and amphiregulin (AREG), two ligands for EGFR, which were significantly attenuated by castration (**P<0.01, ***P<0.001 versus wild-type group; ††P<0.01, †††P<0.001 versus sham Dsk5 group; n=8 in wild-type group and n=7 in other groups).

Article Snippet: Rat anti-mouse F4/80 (MCA497R, a marker of macrophages), CD68 (MCA1957), CD3 (MCA1477, a marker of T lymphocytes), CD4 (MCA2961), CD8a (MCA2694), and Ly-6G (Gr-1, MCA2387, a marker of neutrophils) were purchased from AbD Serotec (now Bio-Rad); goat anti–phospho-EGFR (Tyr 1173, SC-12351) and anti-human connective tissue growth factor (CTGF; SC-14939), rabbit anti–phospho-EGFR (Tyr845, SC-23420-R) and anti–IL-6 (SC-12912T) were purchased from Santa Cruz Biotechnology; rabbit anti-Wilms Tumor Protein (WT1, a marker of podocytes; ab89901) and TNF- α (ab6671) were from Abcam; goat anti-EGFR (E1282) and mouse anti– α -smooth muscle actin ( α -SMA; a marker of myofibroblasts, A5228) were from Sigma (St. Louis, MO); rabbit anti-collagen I (600–401–103–01) was purchased from Rockland Immunochemicals; rat anti–KIM-1 (a marker of kidney epithelial injury, MAB1817–100) and mouse anti–4-hydroxynonenal (a marker of oxidative stress, no. 198960) were from R&D Systems; and rabbit anti–p-ERK (4370S) was from Cell Signaling Technology.

Techniques: Expressing, Activation Assay, Immunofluorescence, Staining, Marker

Journal: Brazilian Journal of Pharmaceutical Sciences

Article Title: Swertiamarin attenuates carbon tetrachloride (CCl4)-induced liver injury and inflammation in rats by regulating the TLR4 signaling pathway

doi: 10.1590/s2175-97902018000417449

Figure Lengend Snippet: FIGURE 4 - STM repressed the protein expression of CTGF, TLR4 and NF-κB p65 in the liver. The CTGF protein level of rats in CCl4+STM 100 mg/kg and CCl4+STM 200 mg/kg groups were downregulated significantly compared with the CCl4 group. The protein expression of TLR4 and NF-κB p65 of rats in CCl4+STM 100 mg/kg and CCl4+STM 200 mg/kg groups were significantly lower than that in the CCl4 group. Data are represented as means ± S.D for 3-4 animals per group. * p < 0.05 versus control, ** p < 0.01 versus control; # p < 0.05 versus CCl4, ## p < 0.01 versus CCl4 by one-way ANOVA and LSD post hoc test.

Article Snippet: Antibodies against connective tissue growth factor (CTGF) (Boster, PB0570), TLR4 (Boster, BA1717), NFκB p65 (Santa Cruz, sc-1173), ser307pIRS-1 (Boster, P00268), GAPDH (Boster, BA2913), and Histone H3 (Santa Cruz, sc-166574) were used in this study.

Techniques: Expressing, Control

Journal: Current eye research

Article Title: Immunohistochemical Detection of CTGF in the Human Eye.

doi: 10.3109/02713683.2016.1143014

Figure Lengend Snippet: Figure 2. (A) CTGF-immunoreactivity (A, red) was detected in the corneal epithelium, mainly concentrated in basal layers (blue: DAPI). (B) Single superficial epithelial cells were detected with rather intense immunoreactivity for CTGF (red, arrowhead; blue: DAPI). (C) CTGF-immunoreactivity (red) was also detectable in corneal endothelial cells (arrows), as well as in keratinocytes of the stroma (arrowheads; blue: DAPI). (D) CTGF-immunoreactivity was absent in corresponding negative controls (here: corneal epithelium and stroma; blue: DAPI). (E, F) Conjunctival epithelial cells displayed immunoreactivity for CTGF (E), but CTGF-immunoreactivity was absent in the corresponding negative control (F). Arrowheads: goblet cells; blue: DAPI). (G, H) In the trabecular meshwork (asterisk) and Schlemm’s canal (arrowheads), CTGF immunoreactivity (red) revealed an identical staining pattern (G) as compared with corresponding negative controls (H).

Article Snippet: After a 5 min rinse, slides were incubated overnight at RT with an anti-human connective tissue growth factor (CTGF; raised in goat, 1:100, AF660, R&D Systems, Minneapolis, MN, USA) in TBS, containing 1% BSA and 0.5% Triton X-100.

Techniques: Negative Control, Staining

Journal: Current eye research

Article Title: Immunohistochemical Detection of CTGF in the Human Eye.

doi: 10.3109/02713683.2016.1143014

Figure Lengend Snippet: Figure 3. (A, B) In the iris, CTGF-immunoreactivity (A, red) was present in anterior layers of the iris (arrows) as well as muscle fibers of the iris sphincter (arrowheads), as detected with alpha-smooth-muscle actin (green), but was absent in the corresponding negative control (B). DAPI: white. (C, D) In C, iris vessels displayed immunoreactivity for CTGF (red) in the vascular endothelium (blue, CD31), as seen by an association of both signals (purple color), while an overlap of CTGF with vascular smooth muscle cells (green, ASMA) was not observed, as seen by absence of yellow-mixed color. Immunoreactivity was absent in corresponding negative controls (D). Asterisks indicates vessel lumen; DAPI: white. (E, F) CTGF-immunoreactivity (E, red) was present in the muscle fibers of the ciliary body (asterisk), as detected with alpha – smooth muscle actin (green), but was absent in the corresponding negative control (E). DAPI: white. (G, H) CTGF-immunoreactivity was present in the non-pigmented ciliary epithelium (G, arrowheads), but was absent in corresponding negative controls (F; red signal here, arrows, corresponds to autofluorescence). DAPI: white. (I, J) CTGF-immunoreactivity (red) in the lens was absent in the lens epithelium (I, arrowheads), but present in superficial layers of the lens (I, asterisk), while it was absent in the corresponding negative control (J, asterisk). DAPI: blue.

Article Snippet: After a 5 min rinse, slides were incubated overnight at RT with an anti-human connective tissue growth factor (CTGF; raised in goat, 1:100, AF660, R&D Systems, Minneapolis, MN, USA) in TBS, containing 1% BSA and 0.5% Triton X-100.

Techniques: Negative Control

Journal: Current eye research

Article Title: Immunohistochemical Detection of CTGF in the Human Eye.

doi: 10.3109/02713683.2016.1143014

Figure Lengend Snippet: Figure 4. (A, B) In the retina, CTGF-immunoreactivity (red) was present in the nerve fiber layer (NFL; A: cross-section close to the optic nerve head), and further a weak signal was also present in the IPL and OPL, while immunoreactivity was absent in corresponding negative controls (F). DAPI: blue. (C, D) In the choroid, CTGF- immunoreactivity (red) was present in the choriocapillaris (C, arrowheads) and in blood vessels of the choroidal stroma (C, arrows), but was absent in corresponding negative controls (D). DAPI: blue; RPE: retinal pigment epithelium; CC: choriocapillaris. (E, F) In the optic nerve head, CTGF-immunoreactivity (red) was present in endothelial cells of the central retinal artery (E, asterisks) and was detected in single cells within connective tissue strands of the optic nerve (F, arrowhead; inset represents magnified situation in F). DAPI: blue. All images in Figures 2, 3, and 4 represent confocal images in single optical section mode.

Article Snippet: After a 5 min rinse, slides were incubated overnight at RT with an anti-human connective tissue growth factor (CTGF; raised in goat, 1:100, AF660, R&D Systems, Minneapolis, MN, USA) in TBS, containing 1% BSA and 0.5% Triton X-100.

Techniques: