ABSTRACT
Objective To investigate the protective effect and mechanism of Danshen Tongluo Jiedu Decoction medicated serum for hypoxia/reoxygenation rat myocardial microvascular endothelial cells(CMECs)by regulating MALAT1.Methods Rats CMECs cells were cultured in vitro to establish a model of hypoxia/reoxygenation damaged cells,and were transfected overexpressing/silencing blank MALAT1 slow virus,cells were divided into overexpressed blank + TCM group,overexpressed MALAT1 + TCM group,overexpressed MALAT1 group,silenced blank + TCM group,silenced MALAT1 group,and silenced MALAT1 + TCM group.They were cultured with corresponding serum separately.Beclin-1 protein expression was detected by immunofluorescence method,and SRPK1,SRSF1,VEGF and Bax protein expressions were detected by Western blot,MALAT1,SRPK1 and SRSF1 mRNA expressions were detected by RT-PCR.Results Compared with the overexpressed blank + TCM group,Beclin-1 protein expression increased in the overexpressed MALAT1 + TCM group,the protein expressions of SRPK1,SRSF1 and Bax significantly increased(P<0.05,P<0.01),VEGF protein expression significantly decreased(P<0.01),while MALAT1,SRPK1 and SRSF1 mRNA expressions significantly increased(P<0.05,P<0.01).Compared with the overexpressed MALAT1 group,the protein expression of Beclin-1 in overexpressed MALAT1 + TCM group decreased,the expressions of SRPK1,SRSF1 and Bax protein significantly decreased(P<0.01,P<0.05),the expression of VEGF protein significantly increased(P<0.01),the mRNA expressions of MALAT1,SRPK1 and SRSF1 significantly decreased(P<0.05).Compared with the silenced blank + TCM group,the protein expression of Beclin-1 in silenced MALAT1 + TCM group decreased,the expressions of SRPK1,SRSF1 and Bax protein significantly decreased(P<0.01),while the expression of VEGF protein significantly increased(P<0.01),the mRNA expressions of MALAT1,SRPK1 and SRSF1 significantly decreased(P<0.01).Compared with the silenced MALAT1 group,the protein expression of Beclin-1 in silenced MALAT1 + TCM group decreased,the expressions of SRPK1,SRSF1 and Bax protein significantly decreased(P<0.05),the expression of VEGF protein significantly increased(P<0.01),the mRNA expressions of MALAT1,SRPK1 and SRSF1 significantly decreased(P<0.01,P<0.05).Conclusion Upregulation of MALAT1 expression can promote autophagy in hypoxia/reoxygenation model CMECs,while Danshen Tongluo Jiedu Decoction medicated serum can inhibit MALAT1 expression,thus inhibiting autophagy and promoting angiogenesis,and the mechanism may be related to the downregulation of SRPK1 and SRSF1 expressions.
ABSTRACT
Objective To discuss the clinical value of protein induced by vitaminK absence antagonist-Ⅱ (PIVKA-Ⅱ)and al-pha-fetoprotein (AFP)in diagnosing primary hepatocellular carcinoma (PHC).Methods There were 178 samples from in-patients in Fuzhou Infectious Disease Hospital,including 54 patients with PHC,39 patients with liver cirrhosis,55 patients with hepatitis and 30 cases of healthy.Serum levels of PIVKA-II and AFP levels were detected by LUMI-PULSEG1200 au-tomatic immunity analyzer and Abbott automatic immunity analyzer respectively,and the difference between the levels was compared.Analyzed the areas under the receiver operating characteristic curves (ROC-AUC)and compared the sensitivity and specificity of single PIVKA-II or AFP assay,and the combined detection of PHC.Results The serum level of PIVKA-Ⅱ in hepatocellular carcinoma group was 274 mAU/ml,which was higher than that in liver cirrhosis group (23 mAU/ml), chronic hepatitis group (26 mAU/ml)and healthy group (21 mAU/ml)(P<0.001),and the levels of AFP in PHC group was 84.0 ng/ml,which was higher than that in liver cirrhosis (21.78 ng/ml)and healthy groups (2.8 ng/ml).But it was not statistically significant (P=0.585)compared with those in the chronic hepatitis group (66.8 ng/ml),the results of re-ceiver operating characteristic (ROC)curve showed that the area under the curve of PIVKA-Ⅱ was 0.776,higher than the AFP (0.649),(Z=2.262,P=0.023 7).Serum PIVKA-Ⅱ (≥40 mAU/ml)had a sensitivity of 78.52% and a specificity of 76.23% in the diagnosis of PHC,While serum AFP (≥10 mg/ml)had a sensitivity of 77.78% and a specificity of 34.64%in the diagnosis of PHC.A combination of serum levels of PIVKA-Ⅱ and AFP could increase the sensitivity in the diagnosis of PHC (vs PIVKA-Ⅱ,P=0.031;vs AFP,P=0.016)and specificity (vs PIVKA-Ⅱ,P=0.004;vs AFP,P=0.001).Con-clusion Serum PIVKA-Ⅱ have high clinical application values in diagnosing PHC.A combination of serum levels of PIV-KA-Ⅱ and AFP could increase the sensitivity and specificity in diagnosis of PHC.