Mechanism of total flavonoids of Rhododendra simsii in alleviating ischemic brain injury.
10.19540/j.cnki.cjcmm.20221010.402
- Author:
Chen-Chen JIANG
1
;
Lei SHI
1
;
Xin-Ya ZHAO
2
;
Hui ZHANG
2
;
Zi-Xu LI
3
;
Jia-Jun LU
2
;
Yu-Xiang HE
4
;
Di CAO
3
;
Hao-Ran HU
5
;
Jun HAN
6
Author Information
1. Third-Grade Pharmacology Laboratory of National,Administration of Traditional Chinese Medicine Wuhu 241002,China School of Pharmacy,Drug Research and Development Center,Wannan Medical College Wuhu 241002,China.
2. Third-Grade Pharmacology Laboratory of National,Administration of Traditional Chinese Medicine Wuhu 241002,China Anhui Provincial Engineering Laboratory for Screening and Re-evaluation of Active Compounds of Herbal Medicines in Southern Anhui Wuhu 241002,China.
3. Anhui Provincial Engineering Laboratory for Screening and Re-evaluation of Active Compounds of Herbal Medicines in Southern Anhui Wuhu 241002,China Anhui Provincial Engineering Research Center for Polysaccharide Drugs Wuhu 241002,China.
4. Anhui Provincial Engineering Laboratory for Screening and Re-evaluation of Active Compounds of Herbal Medicines in Southern Anhui Wuhu 241002,China School of Pharmacy,Drug Research and Development Center,Wannan Medical College Wuhu 241002,China.
5. Third-Grade Pharmacology Laboratory of National,Administration of Traditional Chinese Medicine Wuhu 241002,China Anhui Provincial Engineering Research Center for Polysaccharide Drugs Wuhu 241002,China.
6. Third-Grade Pharmacology Laboratory of National,Administration of Traditional Chinese Medicine Wuhu 241002,China Anhui Provincial Engineering Laboratory for Screening and Re-evaluation of Active Compounds of Herbal Medicines in Southern Anhui Wuhu 241002,China Anhui Provincial Engineering Research Center for Polysaccharide Drugs Wuhu 241002,China School of Pharmacy,Drug Research and Development Center,Wannan Medical College Wuhu 241002,China.
- Publication Type:Journal Article
- Keywords:
SOCE pathway;
STIM-ORAI;
cerebral ischemia-reperfusion injury;
total flavonoids of Rhododendra simsii
- MeSH:
Animals;
Male;
Rats;
Apoptosis;
Brain Ischemia/metabolism*;
Caspase 3;
Interleukin-1;
Interleukin-6;
Rats, Sprague-Dawley;
Reperfusion Injury/metabolism*;
Tumor Necrosis Factor-alpha/genetics*;
Flavonoids/pharmacology*;
Rhododendron/chemistry*
- From:
China Journal of Chinese Materia Medica
2023;48(2):455-464
- CountryChina
- Language:Chinese
-
Abstract:
This study explores the effect of total flavonoids of Rhododendra simsii(TFR) on middle cerebral artery occlusion(MCAO)-induced cerebral injury in rats and oxygen-glucose deprivation/reoxygenation(OGD/R) injury in PC12 cells and the underlying mechanism. The MCAO method was used to induce focal ischemic cerebral injury in rats. Male SD rats were randomized into sham group, model group, and TFR group. After MCAO, TFR(60 mg·kg~(-1)) was administered for 3 days. The content of tumor necrosis factor-α(TNF-α), interleukin-1(IL-1), and interleukin-6(IL-6) in serum was detected by enzyme-linked immunosorbent assay(ELISA). The pathological changes of brain tissue and cerebral infarction were observed based on hematoxylin and eosin(HE) staining and 2,3,5-triphenyltetrazolium chloride(TTC) staining. RT-qPCR and Western blot were used to detect the mRNA and protein levels of calcium release-activated calcium channel modulator 1(ORAI1), stromal interaction molecule 1(STIM1), stromal intera-ction molecule 2(STIM2), protein kinase B(PKB), and cysteinyl aspartate specific proteinase 3(caspase-3) in brain tissues. The OGD/R method was employed to induce injury in PC12 cells. Cells were randomized into the normal group, model group, gene silencing group, TFR(30 μg·mL~(-1)) group, and TFR(30 μg·mL~(-1))+gene overexpression plasmid group. Intracellular Ca~(2+) concentration and apoptosis rate of PC12 cells were measured by laser scanning confocal microscopy and flow cytometry. The effect of STIM-ORAI-regulated store-operated calcium entry(SOCE) pathway on TFR was explored based on gene silencing and gene overexpression techniques. The results showed that TFR significantly alleviated the histopathological damage of brains in MCAO rats after 3 days of admini-stration, reduced the contents of TNF-α, IL-1, and IL-6 in the serum, down-regulated the expression of ORAI1, STIM1, STIM2, and caspase-3 genes, and up-regulated the expression of PKB gene in brain tissues of MCAO rats. TFR significantly decreased OGD/R induced Ca~(2+) overload and apoptosis in PC12 cells. However, it induced TFR-like effect by ORAI1, STIM1 and STIM2 genes silencing. However, overexpression of these genes significantly blocked the effect of TFR in reducing Ca~(2+) overload and apoptosis in PC12 cells. In summary, in the early stage of focal cerebral ischemia-reperfusion injury and OGD/R-induced injury in PC12 cells TFR attenuates ischemic brain injury by inhibiting the STIM-ORAI-regulated SOCE pathway and reducing Ca~(2+) overload and inflammatory factor expression, and apoptosis.
