| Objective:To evaluate the role of heat shock protein 22?HSP22?in atherosclerosis?AS?induced by high-fat diet and in intervention with atorvastatin?ator?.Methods:1.Total 3 groups of 8-9 weeks-old Apo E-/-,HSP22-/-//Apo E-/-,HSP22+//Apo E-/-male mice were used,sheach group were 18 mice.After one week of adaptive feeding,the mice in each group was randomly divided into 2 subgroups:control group and ator group,HSP22 knockout group?KO group?and HSP22 knockout ator group?KO+ator group?,HSP22 overexpression group?Tg group?and HSP22 overexpression ator group?Tg+ator group?were set up.All these mice were fed with high fat diet for 12weeks to establish atherosclerosis models.Atro(ator 10mg/kg-1-d-1)was administered to mice of ator group,KO+ator group and Tg+ator group from the 5th week.Other groups were administered by saline.2.The weight of all mice was recorded every week.The lipid levels were measured at baseline and the end of intervention.3.The Oil Red O staining and HE staining of the aortic wall of the mice were used to measure the atherosclerotic lesion burdens.4.The protein levels of HSP22,NF-?B,e NOS,ICAM-1 and IL-6 in the aortas or serum were examined by Western blotting or immunohist ochemistry?IHC?or ELISA.Results:1.The changes of lipid level?1?The baseline lipid shows no difference in all groups?P>0.05?;?2?The lipid at the end of intervention:there were no significant difference in serum TC,TG and HDL-C of mice among the control group,KO group and Tg group?P>0.05?.LDL-C in Tg group was significantly less than KO group?13.59±0.84vs15.41±0.39,P<0.01?.After 8 weeks of ator intervention,there were no difference in serum TC,TG,LDL-C and HDL-C of mice among the ator group,KO+ator group and Tg+ator group?P>0.05?.2.Histopathological observationAortic Oil Red O staining showed the relative area of aorta plaque in Tg group was less than that in KO group??12.68±0.75?%vs?13.75±0.56?%,P<0.05?.After 8weeks of ator intervention,the relative area of aorta plaque in Tg+ator group was significantly less than that in KO+ator group??3.78±0.20?%vs?4.90±0.65?%,P<0.01?.3.The protein expression of HSP22,e NOS,NF-?B and ICAM-1 were measured by WB or IHCThe protein expression of HSP22 and e NOS in Tg group was significantly higher than that in control group?HSP22:1.08±0.06vs0.31±0.14,P<0.01?and KO group?HSP22:1.08±0.06vs0.06±0.04,P<0.01;e NOS:0.40±0.03vs0.21±0.01,P<0.01?,and their expression in control group was significantly higher than that in KO group?HSP22:0.31±0.14vs0.06±0.04,P<0.01;e NOS:0.39±0.02vs0.21±0.01,P<0.01?.The pro tein expression of NF-?B and ICAM-1 in control group was significantly less compared with KO group?NF-?B:0.46±0.02vs0.52±0.02,P<0.01;ICAM-1:0.34±0.01vs0.74±0.02,P<0.01?,and their expression was significantly higher than that in Tg gro up?NF-?B:0.46±0.02vs0.29±0.01,P<0.01;ICAM-1:0.34±0.01vs0.18±0.01,P<0.01?.After 8 weeks of ator intervention,the protein expression of HSP22 and e NOS in Tg+ator group was significantly higher than that in ator group?HSP22:0.70±0.03vs0.13±0.02,P<0.01?and KO+ator group?HSP22:0.70±0.03vs0.03±0.02,P<0.01;e NOS:0.47±0.02vs0.35±0.03,P<0.01?,and their expressions in ator group were significantly higher than KO+ator group?HSP22:0.13±0.02vs0.03±0.02,P<0.01;e NOS:0.46±0.02vs0.35±0.03,P<0.01?.The expressions of NF-?B and ICAM-1 protein in KO+ator group were significantly higher than ator group and Tg+ator group?ICAM-1:0.70±0.03vs0.23±0.02or0.04±0.01,P<0.01;NF-?B:0.39±0.02vs0.17±0.01or0.15±0.01,P<0.01?,and the protein expression of ICAM-1 in Tg+ator group was significantly less than that in ator group?ICAM-1:0.04±0.01vs0.23±0.02,P<0.01?.4.The secretions of HSP22 and IL-6 from serum samples of all groups were measured by ELISAThe concentration of serum HSP22 in KO group was significantly less than that in control group and Tg group?45.84±1.18vs181.37±21.21or248.17±36.78,P<0.01?,and its expression in control group was significantly less than that in Tg group?181.37±21.21vs 248.17±36.78,P<0.01?.No difference in concentration of serum IL-6 was found between Tg group,KO group and control group?271.43±12.13vs263.62±11.58or 251.29±39.98,P>0.05?.After 8 weeks of ator intervention,the concentration of serum HSP22 in KO+ator group was significantly less than that in ator group and Tg+ator group?38.97±8.36vs145.83±6.95or203.28±18.82,P<0.01?,and its expression in Tg+ator group was significantly higher than that in ator group?203.28±18.82vs145.83±6.95,P<0.01?.The concentration of serum IL-6 between Tg+ator group,KO+ator group and ator group was not different?160.16±6.27vs172.44±9.48or144.95±11.98,P>0.05?.Conclusion:1.High fat feeding induces the expression of NF-?B,ICAM-1 and IL-6,and inhibits the expression of e NOS,accelerates AS.The intervention with ator up-regulates the expression of e NOS and down-regulates the expression of NF-?B,IL-6 and ICAM-1,thus attenuating AS development.2.HSP22 gene deletion up-regulates the expression of NF-?B and ICAM-1,and down-regulates the expression of e NOS,accelerates AS.HSP22 overexpression decreases the expression of NF-?B and ICAM-1,thus attenuating AS development.3.HSP22 gene deletion partial limits the role of ator on the expression of NF-?B,ICAM-1 and e NOS.HSP22 overexpression amplifies the reduced expression of ICAM-1 by the intervention with ator,further improveattenuates AS development.Key words:atherosclerosis;heat shock protein 22;atorvastatin;inflammation. |
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