ABSTRACT
Objective To explore the acute toxicity effect of CKLF1-C19,a polypeptide of chemokine-like factor 1(CKLF1),on the KM mice. Methods A total of 40 KM mice,half male and half female,were randomly divided into 2 groups. The mice in the experimental group were injected with CKLF1-C19 at a dose of 25 mL/kg(100 μg/mL,1 mg/mL and 10 mg/mL)through the tail vein,and those in the control group received an equal volume of sterile saline solution. Changes in the body weight of the mice were recorded the day after treatment, and the general conditions of mice in the experimental group were observed closely and compared with the normal group. Then blood samples were taken from the abdominal aorta to measure liver and kidney function. Tissue samples of liver, kidney, spleen and lung were taken for histopathological examination by HE staining. Results In the maximum tolerance test,the mice of the two groups were in good condition, and their body weight was increased gradually, without significant difference between the experimental group and the control group(P > 0.05). There was no death within 14 days. The blood biochemical indexes of liver and kidney function showed no significant differences between the two groups(P > 0.05). The gross appearances of heart, liver,kidney,spleen and lung were normal in the two mouse groups, and the pathological examination with HE staining showed normal clear structure with no obvious changes in these organs of each group. Conclusions Our results demonstrate that CKLF1-C19 has no acute toxicity effect on mice.
ABSTRACT
AIM:To investigate the effect of CKLF1-C19 polypeptide (C19) on differentiation of human lung fibroblast (LFB) into myofibroblast (MFB) induced by TGF-β. METHODS:LFBs were cultured and identified. LFBs were treated with TGF-β(5 μg/L) to establish the cell model of LFB differentiate into MFB. The LFBs were divided into 6 experimental groups including control group,TGF-β group,and TGF-β plus different doses(1,0.1,0.01,0.001 mg/L) C19 groups. The cell morphology,cell proliferation rate, and the expression of α smooth muscle actin (α-SMA) and collagen Ⅰ were observed. RESULTS:Human primary LFB was successfully cultured and was confirmed by the method of immunofluorescence. TGF-β at 5 μg/L induced proliferation and differentiation of LFB. The mRNA levels of α-SMA and collagen Ⅰ in TGF-β group were higher than that in control group(P<0.05).The cell proliferation rates,mRNA levels of α-SMA and collagen Ⅰ, and the protein expression of α-SMA in 0.01 mg/L+TGF-β group and 0.001 mg/L+TGF-β group were markedly lower than those in TGF-β group(P<0.05). CONCLUSION:C19 at 0.01 mg/L and 0.001 mg/L effectively inhibits differentiation of LFB into MFB induced by TGF-β, thus inhibiting the process of airway remodeling and fibrosis to some extent.
ABSTRACT
Aim To investigate the role of chemokine-like factor 1 ( CKLF1 ) in SH-SY5 Y cell migration and its molecular regulatory mechanism. Methods SH-SY5Y cells were stimulated with CKLF1 for 0. 5 h, 2 h, 8 h and 24 h, respectively. The migration distance and the percentage of migration cells were recorded by CELLocate analysis. The phosphorylation of focal ad-hesion kinase ( FAK) at Tyr-397 site was detected by Western blot analysis. By chemotaxis assays, we con-firmed the chemotaxis of CKLF1. Furthermore, FAK inhibitor PF-573228 and PLCγ inhibitor U73122 were used for the research of molecular regulatory mecha-nisms involved. Results CKLF1 promoted cell migra-tion and induced a strong increase in the phosphoryla-tion level of FAK-pY397 , which were significantly at-tenuated by the presence of U73122 ( a specific inhibi-tor for PLCγ) . In addition, the chemotaxis of CKLF1 was obviously blocked by the FAK inhibitor PF-573228 . Conclusion CKLF1 induces SH-SY5 Y cell migration via PLCγ/FAK signaling pathway.