XingNaoJing, prescription of traditional Chinese medicine, prevents autophagy in experimental stroke by repressing p53-DRAM pathway

Abstract

BackgroundXingnaojing (XNJ), a well known prescription in traditional Chinese medicine, has been used for treatment of stroke in China. However, the effects and mechanisms of XNJ on autophagy are not clear. Here, we used the cell models of autophagy induced by serum-free condition and ischemia stroke in rats to further investigate whether the p53-DRAM pathway is involved in the effects of XNJ on autophagy.MethodsWe used the cell model of autophagy induced by serum-free condition and the rat model of ischemia caused by a middle cerebral artery occlusion (MCAO). The effects of XNJ on p53 transcriptional activity of PC12 cells were evaluated by the luciferase activity assay. The mRNA levels and the expression of p53 and its target autophagy gene DRAM (damage-regulated autophagy modulator) were analyzed respectively by Quantitative-RTPCR and Western blot assay. The activation of autophagy was detected by the levels of autophagy markers, microtubule associated protein light chain 3 (LC3) and p62 by Immunofluorescence and Western blot. p53 inhibitor was used to determine whether p53 is responsible for the effects of XNJ on preventing autophagy.ResultsThe assay for luciferase activity of p53 promoter indicated that XNJ inhibited p53 transcriptional activity. XNJ reduced the expression of p53 and its target autophagy gene DRAM (damage-regulated autophagy modulator) in serum-free condition PC12 cells and the cortex in MCAO rats. XNJ reduced autophagy of PC12 cells induced by serum-free condition and the cortex in MCAO rats. Furthermore, suppression of p53 by p53 inhibitor significantly reduced the effects of XNJ on the autophagy of PC12 cells in serum-free condition.ConclusionXNJ prevents autophagy in experimental stroke by repressing p53/DRAM pathway. Our findings are therefore of considerable therapeutic significance and provide the novel and potential application of XNJ for the treatment of brain diseases.

Tanshinone I induces cell apoptosis by reactive oxygen species-mediated endoplasmic reticulum stress and by suppressing p53/DRAM-mediated autophagy in human hepatocellular carcinoma

“…Hsieh et al reported that sedanolide promotes autophagy via the p53/DRAM signaling pathway in human liver cancer J5 cells [39]. Wei et al also demonstrated that XingNaoJing inhibits autophagy by suppressing the p53/ DRAM signaling pathway in PC12 cells [12]. Interestingly, our results demonstrated that TA I decreased p53 and DRAM expression in HepG2 and Huh7 cells.…”

“…Accumulated evidence suggests that autophagy plays an important role in cell apoptosis mediated by anti-cancer drugs [9,10]. Furthermore, the p53/damage-regulated autophagy modulator (DRAM) signaling pathway has been reported to be involved in regulating autophagy [11,12]. Many studies have revealed that ROS accumulation in various types of cancers involves cell apoptosis and autophagy induced by anti-tumor drugs [13,14].…”

Single standard substance for the determination of nine volatile components in the distillate of Fructus Gardeniae and Radix Curcumae (an intermediate of Xingnaojing Injection)

“…For another component x, A x was the peak area acquired from m x amount of component x, and the correction factor (f x ) was calculated as m x /A x . Then, the RCF of x to s (f x/s ) could be calculated by formula (1).…”

“…Xingnaojing Injection is a traditional Chinese medicine prepared from Moschus , Fructus Gardeniae (FG), Radix Curcumae (RC), and borneol and is clinically used for treating stroke , cerebral ischemia , consciousness disturbance , and alcohol intoxication , with annual sales of more than two billion Chinese yuan. However, some adverse effects of Xingnaojing Injection have also been reported , which necessitates more strict QC of this injection.…”

Xingnaojing for Moderate-to-severe Acute ischemic Stroke (XMAS): study protocol for a randomized controlled trial

“…XNJ consists of four Chinese herbs: Moschus , Radix curcumae , borneol , and Fructus gardeniae . Previous studies have demonstrated that XNJ may has multiple neuroprotective mechanisms in terms of inhibiting glutamate-induced apoptosis [ 7 ], anti-autophagy via the p53-DRAM signaling pathway [ 8 ], decreasing gamma-aminobutyric acid (GABA) [ 9 ], improving neurobehavioral disturbances, as well as reducing infarct size [ 10 ]. Recently, a systematic review and meta-analysis to assess the efficacy of XNJ for stroke demonstrated that XNJ may reduce brain injury and improve functional recovery after stroke.…”