Exosomal miRNA-222 Alerts shRNA-PCSK9 Induction Brain Tau Hyperphosphorylation
10.13748/j.cnki.issn1007-7693.20222249
- VernacularTitle:外泌体miRNA-222预警shRNA-PCSK9诱发脑Tau蛋白过度磷酸化
- Author:
Lei WANG
1
;
Qian JIANG
1
;
Hong WANG
1
;
Ling YUAN
1
;
Nan LYU
1
;
Di HAO
1
;
Xiaoxue CUI
1
;
Zi WANG
1
Author Information
1. Cardiovascular and Cerebrovascular Drugs Research and Development Center, Tianjin Institute of Medical and Pharmaceutical Sciences, Tianjin 300020, China
- Publication Type:Journal Article
- Keywords:
hypercholesterolemia ； PCSK9； cognitive impairment ； miRNA-222
- From:
Chinese Journal of Modern Applied Pharmacy
2024;41(5):636-643
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE :To investigate whether the microRNA-222(miRNA-222) carried by plasma exosomes can serve as an early warning marker for cognitive impairment induced by shRNA-PCSK9.
METHODS
The high-fat diet(HFD) was used to prepare a hypercholesterolemic mouse model group. The model group mice were divided into HFD-shRNA control group and HFD-shRNA-PCSK9 group. The shRNA-PCSK9 was constructed, injected intravenously into the body, and the expression of PCSK9 mRNA was detected by real-time PCR(RT-PCR). Tau protein and phosphorylation in brain tissue were observed by immunohistochemistry (IHC). Western blotting was used to detect Tau protein and P-Tau protein. Serum amyloid Aβ1-42Ab levels were determined by ELISA. The kits extracted plasma exosomes step by step, identify the exosome morphology by negative staining electron microscopy, and determined the size of exosomes by NTA technology. RT-PCR technique was used to detect the expression level of miRNA-222 carried in plasma exosomes.
RESULTS
The model mouse were prepared by feeding HFD for 13 weeks, whose total cholesterol(TC) and low-density lipoprotein(LDL-C) contents in serum were significantly increased. At the same time, the expression of PCSK9 mRNA in the brain tissue of model group was significantly increased. After shRNA-PCSK9 lentivirus interference, PCSK9 mRNA expression was inhibited, and IHC observed that shRNA-PCSK9 induced abnormal expression and hyperphosphorylation of Tau protein in brain tissue, indicating that the pathological changes of neurofibrillary tangles had occurred. However, at this time, serum Aβ1-42Ab had not been significantly increased, and it had not yet been of significance for the diagnosis of cognitive impairment. The miRNA in plasma exosomes was extracted, and RT-PCR results showed that the expression of miRNA-222 carried in the exosomes of the HFD-shRNA-PCSK9 group was significantly lower than that of the HFD-shRNA control group.
CONCLUSION
Plasma exosomes carried miRNA-222 provides an early warning marker for shRNA-PCSK9- induced cognitive impairment.
