中国病理生理杂志
中國病理生理雜誌
중국병리생리잡지
CHINESE JOURNAL OF PATHOPHYSIOLOGY
2014年
4期
577-583
,共7页
张运%王莉莉%赵茜%马士程%贺茂林
張運%王莉莉%趙茜%馬士程%賀茂林
장운%왕리리%조천%마사정%하무림
G蛋白偶联受体激酶5%NF-κB%星形细胞%炎症%氧化性应激
G蛋白偶聯受體激酶5%NF-κB%星形細胞%炎癥%氧化性應激
G단백우련수체격매5%NF-κB%성형세포%염증%양화성응격
G-protein-coupled receptor kinase 5%NF-kappa B%Astrocytes%Inflammation%Oxidative stress
目的：探讨培养新生大鼠皮层星形胶质细胞G蛋白偶联受体激酶5（ GRK5）基因沉默后核因子κB （ NF-κB）表达的变化及其与胶质细胞活化、炎症反应和氧化应激的关系。方法：采用分别或联合RNA干扰沉默GRK5基因表达与NF-κB抑制剂N-乙酰半胱氨酸干预进行实验分组。利用免疫荧光、实时定量PCR和Western blotting观察GFAP与活性caspase-3表达，细胞分泌TNF-α与NO的浓度， p65、TNF-α、IL-1β和iNOS的表达情况等。结果：GRK5 siRNA刺激星形胶质细胞活化，并检测到NF-κB表达增多（P＜0.01），TNF-α、IL-1β和iNOS mR-NA表达水平增高（P＜0.01），细胞分泌TNF-α和NO增多（P＜0.01），活性caspase-3表达增多（P＜0.01）。采用GRK5 siRNA＋NF-κB抑制剂联合干预可部分逆转GRK5 siRNA引起的上述变化（ P＜0.05）。结论：GRK5基因沉默可能通过刺激NF-κB表达增多引起星形胶质细胞活化。GRK5的正常表达可能具有抑制星形胶质细胞活化相关炎症反应及氧化应激的作用。
目的：探討培養新生大鼠皮層星形膠質細胞G蛋白偶聯受體激酶5（ GRK5）基因沉默後覈因子κB （ NF-κB）錶達的變化及其與膠質細胞活化、炎癥反應和氧化應激的關繫。方法：採用分彆或聯閤RNA榦擾沉默GRK5基因錶達與NF-κB抑製劑N-乙酰半胱氨痠榦預進行實驗分組。利用免疫熒光、實時定量PCR和Western blotting觀察GFAP與活性caspase-3錶達，細胞分泌TNF-α與NO的濃度， p65、TNF-α、IL-1β和iNOS的錶達情況等。結果：GRK5 siRNA刺激星形膠質細胞活化，併檢測到NF-κB錶達增多（P＜0.01），TNF-α、IL-1β和iNOS mR-NA錶達水平增高（P＜0.01），細胞分泌TNF-α和NO增多（P＜0.01），活性caspase-3錶達增多（P＜0.01）。採用GRK5 siRNA＋NF-κB抑製劑聯閤榦預可部分逆轉GRK5 siRNA引起的上述變化（ P＜0.05）。結論：GRK5基因沉默可能通過刺激NF-κB錶達增多引起星形膠質細胞活化。GRK5的正常錶達可能具有抑製星形膠質細胞活化相關炎癥反應及氧化應激的作用。
목적：탐토배양신생대서피층성형효질세포G단백우련수체격매5（ GRK5）기인침묵후핵인자κB （ NF-κB）표체적변화급기여효질세포활화、염증반응화양화응격적관계。방법：채용분별혹연합RNA간우침묵GRK5기인표체여NF-κB억제제N-을선반광안산간예진행실험분조。이용면역형광、실시정량PCR화Western blotting관찰GFAP여활성caspase-3표체，세포분비TNF-α여NO적농도， p65、TNF-α、IL-1β화iNOS적표체정황등。결과：GRK5 siRNA자격성형효질세포활화，병검측도NF-κB표체증다（P＜0.01），TNF-α、IL-1β화iNOS mR-NA표체수평증고（P＜0.01），세포분비TNF-α화NO증다（P＜0.01），활성caspase-3표체증다（P＜0.01）。채용GRK5 siRNA＋NF-κB억제제연합간예가부분역전GRK5 siRNA인기적상술변화（ P＜0.05）。결론：GRK5기인침묵가능통과자격NF-κB표체증다인기성형효질세포활화。GRK5적정상표체가능구유억제성형효질세포활화상관염증반응급양화응격적작용。
AIM: To study the effect of G-protein-coupled receptor kinase 5 (GRK5) on the activation of astrocytesin the brain cortex of newborn Wistar rats .METHODS: GRK5 gene was silenced in the model of rat brain cortexastrocytes in vitro for 24 h.N-acetylcysteine (NAC), which is a known inhibitor of NF-κB, was added into the culture mediumaccording to gene silencing for 24 h.The expression levels of GFAP and caspase-3 were detected by the method of immunofluorescence,and the mRNA levels of NF-κB, TNF-α, IL-1βand iNOS were determined by real-time PCR.Moreover,the activity of SOD and concentrations of TNF -αand NO were measured.RESULTS: GRK5 gene silencing increasedthe expression of NF-κB at mRNA and protein levels obviously (P <0.01), and the mRNA levels of IL-1βand iNOS increasedsynchronously (P <0.01).Furthermore, caspase-3-positive cells in GRK5 siRNA group were increased comparedwith control siRNA group (P <0.01).Treatment with NAC obviously reduced the activity of NF -κB and weakened theeffects induced by GRK5 siRNA (P <0.05).CONCLUSION: GRK5 siRNA increases NF-κB activity and induces the activationof astrocytes.
