Annao Pingchong decoction alleviate the neurological impairment by attenuating neuroinflammation and apoptosis in intracerebral hemorrhage rats.

Intracerebral hemorrhage (ICH) is a central nervous system disease that causes severe disability or death. Even though Annao Pingchong decoction (ANPCD), a traditional Chinese decoction, has been used clinically to treat ICH in China, its molecular mechanism remains unclear. To study whether the neuroprotective effect of ANPCD on ICH rats is achieved by alleviating neuroinflammation. This paper mainly explored whether inflammation-related signaling pathways (HMGB1/TLR4/NF-κB P65) plays a role in ANPCD treatment of ICH rats. Liquid chromatography-tandem mass spectrometry was used to analyze the chemical composition of ANPCD. ICH models were established by injecting autologous whole blood into the left caudate nucleus of Sprague-Dawley (SD) rats. Modified neurological severity scoring (mNSS) was used to assess the neurological deficits. The levels of tumor necrosis factor (TNF)-α, interleukin (IL)-1β, and IL-6 were analyzed using enzyme-linked immunosorbent assay (ELISA). Pathological changes in the rat brains were observed using hematoxylin-eosin, Nissl, and TUNEL staining. The protein levels of HMGB1, TLR4, NF-κB p65, B-cell lymphoma 2 (Bcl-2), and Bcl-2-associated X protein (Bax) were measured by western blotting and immunofluorescence analysis. Ninety-three ANPCD compounds were identified, including 48 active plasma components. Treatment with ANPCD effectively improved the outcome, as observed by the neurological function scores analysis and brain histopathology. Our results showed that ANPCD exerts its anti-inflammatory effects by significantly downregulating the expression of HMGB1, TLR4, NF-κB p65, TNF-α, IL-1β, and IL-6. ANPCD also exerted anti-apoptotic effects by significantly decreasing the apoptosis rate and Bax/Bcl-2 ratio. We found that ANPCD had neuroprotective effect in clinical work. Here, we also found that the action mechanism of ANPCD might be related to attenuate neuroinflammation and apoptosis. These effects were achieved by inhibiting the expression of HMGB1, TLR4 and NF-κB p65.

Guo C ，Zhou X ，Wang X ，Wang H ，Liu J ，Wang J ，Lin X ，Lei S ，Yang Y ，Liu K ，Long H ，Zhou D ... -  《-》

Integrated Network Pharmacology and in vivo Experimental Validation Approach to Explore the Potential Antioxidant Effects of Annao Pingchong Decoction in Intracerebral Hemorrhage Rats.

Annao Pingchong decoction (ANPCD) is a traditional Chinese decoction which has definite effects on treating intracerebral hemorrhage (ICH) validated through clinical and experimental studies. However, the impact of ANPCD on oxidative stress (OS) after ICH remains unclear and is worth further investigating. To investigate whether the therapeutic effects of ANPCD on ICH are related to alleviating OS damage and seek potential targets for its antioxidant effects. The therapeutic candidate genes of ANPCD on ICH were identified through a comparison of the target genes of ANPCD, target genes of ICH and differentially expressed genes (DEGs). Protein-protein interaction (PPI) network analysis and functional enrichment analysis were combined with targets-related literature to select suitable antioxidant targets. The affinity between ANPCD and the selected target was verified using macromolecular docking. Subsequently, the effects of ANPCD on OS and the selected target were further investigated through in vivo experiments. Forty-eight candidate genes were screened, in which silent information regulator sirtuin 1 (SIRT1) is one of the core genes that has antioxidant effects and ICH significantly affected its expression. The good affinity between 6 compounds of ANPCD and SIRT1 was also demonstrated by macromolecular docking. The results of in vivo experiments demonstrated that ANPCD significantly decreased modified neurological severity scoring (mNSS) scores and serum MDA and 8-OHdG content in ICH rats, while significantly increasing serum SOD and CAT activity, complicated with the up-regulation of ANPCD on SIRT1, FOXO1, PGC-1α and Nrf2. Furthermore, ANPCD significantly decreased the apoptosis rate and the expression of apoptosis-related proteins (P53, cytochrome c and caspase-3). ANPCD alleviates OS damage and apoptosis after ICH in rats. As a potential therapeutic target, SIRT1 can be effectively regulated by ANPCD, as are its downstream proteins.

Zhou X ，Wang X ，Li J ，Zhang M ，Yang Y ，Lei S ，He Y ，Yang H ，Zhou D ，Guo C ... -  《-》

MicroRNA-93 regulates the neurological function, cerebral edema and neuronal apoptosis of rats with intracerebral hemorrhage through TLR4/NF-κB signaling pathway.

Shang Y ，Dai S ，Chen X ，Wen W ，Liu X ... -  《-》

[Ligusticum cycloprolactam inhibits IL-1β-induced apoptosis and inflammation of rat chondrocytes via HMGB1/TLR4/NF-κB signaling pathway].

Qi X ，Chen X ，An WB ，Xu ZM ，Wang DX ，Luo PF ，Chen YX ，Ma JJ ，Hu ZY ，Qi W ，Liu JJ ，Liu JX ... -  《-》

HMGB1/TLR4 induces autophagy and promotes neuroinflammation after intracerebral hemorrhage.

Intracerebral hemorrhage (ICH) causes autophagy as well as inflammation; the latter is known to involve the high-mobility group box 1 protein (HMGB1)/Toll-like receptor 4 (TLR4) axis. Here we investigated whether this axis may help mediate both the autophagy and inflammation associated with ICH. ICH was induced by injecting autologous blood into Sprague-Dawley rats, followed in some cases by intracerebroventricular injection of short interfering RNA (siRNA) against HMGB1 or TLR4 at 6 h after ICH induction or by intraperitoneal injection of the autophagy inhibitor 3-methyladenine (3-MA) or autophagy activator rapamycin at 6, 24, and 48 h after ICH induction. Western blotting, immunohistochemistry or immunofluorescence was used to assess levels of HMGB1/TLR4 signaling pathway proteins as well as markers of autophagy (LC3B, Beclin1, Atg5) or inflammation (IL-1 beta, TNF-α). Numbers of apoptotic cells were determined using TUNEL staining. Changes in levels of these proteins were correlated with neurological deficits measured using the modified Neurological Severity Score. ICH caused HMGB1 to translocate from the nucleus into the cytoplasm, and it up-regulated expression of TLR4 and myeloid differentiation factor 88 (MyD88), and induced neurological deficits. Administering siRNA against HMGB1 or TLR4 reversed this up-regulation. Levels of markers of autophagy (LC3B, Beclin1, Atg5) or inflammation (IL-1 beta, TNF-α) were significantly higher 72 h after ICH than at baseline, as were the numbers of TUNEL-positive cells. Administering siRNA against HMGB1 or TLR4 markedly alleviated inflammation, and autophagy, apoptosis, and neurological deficits. Similarly, administering autophagy inhibitor 3-MA alleviated inflammation, apoptosis, and neurological deficits. Conversely, autophagy activator rapamycin exacerbated these effects of ICH. During the acute phase of ICH, the HMGB1/TLR4/MyD88 axis acts via autophagy to promote inflammation.

Lei C ，Li Y ，Zhu X ，Li H ，Chang X ... -  《-》