Relationship Between NLRP3 Inflammasome and Ischemic Stroke and Intervention Effect of Traditional Chinese Medicine / 中国实验方剂学杂志

Ischemic stroke， also known as cerebral infarction， is the most common type of stroke. Ischemic stroke is extremely harmful with high rates of morbidity， incidence， disability， and mortality， bringing a huge burden on society and families. As a result， finding new and effective prevention and treatment methods is critical. The pathological mechanism of ischemic stroke is very complex and superimposed， with inflammatory response serving as a critical pathological link in the ischemic stroke cascade injury process. NOD-like receptor 3 （NLRP3） is an intracellular sensor， and the inflammatory cascade mediated by the activated NLRP3 inflammasome can exacerbate ischemic stroke injury through the release of inflammatory factors. Taking the NLRP3 inflammasome as the entry point， a large number of experimental studies on the intervention of traditional Chinese medicine （TCM） in the assembly and activation of NLRP3 inflammasome have been carried out， which proved that Chinese medicinal monomers or prescriptions with the main functions of tonifying deficiency， clearing heat and removing toxin， eliminating phlegm， promoting circulation and resolving stasis can interfere with the assembly and activation of NLRP3 inflammasome， reduce the inflammatory response， and relieve ischemic stroke. This study reviewed the assembly and activation of NLRP3 inflammasome， the mechanism of NLRP3 inflammasome in ischemic stroke， and the prevention and treatment of ischemic stroke by TCM through regulation of the NLRP3 inflammasome， which provides a new entry point for the pathological mechanism of ischemic stroke and a direction for the development of new treatments for ischemic stroke.

Relationship Between NLRP3 Inflammasome and Ischemic Stroke and Intervention Effect of Traditional Chinese Medicine / 中国实验方剂学杂志

Ischemic stroke， also known as cerebral infarction， is the most common type of stroke. Ischemic stroke is extremely harmful with high rates of morbidity， incidence， disability， and mortality， bringing a huge burden on society and families. As a result， finding new and effective prevention and treatment methods is critical. The pathological mechanism of ischemic stroke is very complex and superimposed， with inflammatory response serving as a critical pathological link in the ischemic stroke cascade injury process. NOD-like receptor 3 （NLRP3） is an intracellular sensor， and the inflammatory cascade mediated by the activated NLRP3 inflammasome can exacerbate ischemic stroke injury through the release of inflammatory factors. Taking the NLRP3 inflammasome as the entry point， a large number of experimental studies on the intervention of traditional Chinese medicine （TCM） in the assembly and activation of NLRP3 inflammasome have been carried out， which proved that Chinese medicinal monomers or prescriptions with the main functions of tonifying deficiency， clearing heat and removing toxin， eliminating phlegm， promoting circulation and resolving stasis can interfere with the assembly and activation of NLRP3 inflammasome， reduce the inflammatory response， and relieve ischemic stroke. This study reviewed the assembly and activation of NLRP3 inflammasome， the mechanism of NLRP3 inflammasome in ischemic stroke， and the prevention and treatment of ischemic stroke by TCM through regulation of the NLRP3 inflammasome， which provides a new entry point for the pathological mechanism of ischemic stroke and a direction for the development of new treatments for ischemic stroke.

Effect of Yinxing Mihuan Oral Solutionon P2RX7/NLRP3 Signaling Pathway in Depression Model Rats Induced by Isolation Combined with Chronic Unpredictable Mild Stress / 中国实验方剂学杂志

Objective:To explore the antidepressant mechanism of Yinxing Mihuan oral solution （YMO） by investigating its effect on depression model rats. Method:The depression rats were induced by isolation combined with chronic unpredictable mild stress （CUMS） and then randomly divided into model group， fluoxetine group （10 mg·kg^-1） and high-dose （618 mg·kg^-1） and low-dose （309 mg·kg^-1） YMO groups. A blank control group was also set up and ten rats were included in each group. Modeling lasted for 21 consecutive days， and rats were administered the 8th day after stimulation at a dose of 10 mL·kg^-1 for 14 days， except those in the blank control and model groups which were given distilled water. Afterward， the sucrose preference test， open field test， tail suspension test were carried out. The pathological changes of hippocampus in depression rats were observed after hematoxylin-eosin （HE） staining. The content of interleukin-1<italic>β </italic>（IL-1<italic>β</italic>）， interleukin-6 （IL-6） and tumor necrosis factor-<italic>α </italic>（TNF-<italic>α</italic>） in the hippocampus of rats in each group and the expression of NOD-like receptor 3 （NLRP3） and other proteins in its related activation signaling pathways were detected with multi-factor detection （Luminex） and Western blot. Result:After 14 days of continuous administration， compared with the blank control group， the model group witnessed significantly reduced sugar water consumption rate and the times of rearing and significantly prolonged cumulative time of immobility during tail suspension （<italic>P</italic><0.05， <italic>P</italic><0.01）. Compared with the model group， the fluoxetine group and the high-dose YMO group saw increases in the times of rearing， times of crossing and sugar water consumption rate and a significant decrease in the cumulative time of immobility during tail suspension （<italic>P</italic><0.05， <italic>P</italic><0.01）. The results of HE staining showed that the neurons in the hippocampus of rats in the high-dose YMO group were arranged in order and slightly loosened， without obvious microglia infiltration observed. The levels of IL-1<italic>β</italic>， IL-6 and TNF-<italic>α</italic> in the hippocampus of the model group increased significantly as compared with the blank control group （<italic>P</italic><0.05， <italic>P</italic><0.01）， and their content in the high-dose YMO group was significantly lowered in the comparison with the model group （<italic>P</italic><0.05， <italic>P</italic><0.01）. Molecular biology experiments demonstrated that compared with the results of blank group， the expression of purinergic receptor P2X7 （P2RX7）， NLRP3， apoptosis-associated speck-like protein （ASC）， Caspase-1 and IL-1<italic>β</italic> remarkably increased in the model group （<italic>P</italic><0.05， <italic>P</italic><0.01）. Additionally， the expression of P2RX7， NLRP3， ASC， Caspase-1 and IL-1<italic>β </italic>was significantly inhibited in the fluoxetine group and the high-dose YMO group compared with the model group （<italic>P</italic><0.05， <italic>P</italic><0.01）. Conclusion:YMO can improve the depression-like behaviors of rats induced by isolation combined with CUMS， and its mechanism of action is related to the regulation of the P2RX7/NLRP3 signaling pathway.

Effect of Modified Shuyuwan on Neuroinflammation in APP/PS1 Dementia Mice Based on Nlrp3/ASC/Caspase-1 Signaling Pathway / 中国实验方剂学杂志

Objective:To observe the effect of modified Shuyuwan in amyloid precursor protein/ presenilin 1 （APP/PS1） dementia mice on cognitive and memory impairment and to explore its mechanism. Method:The 40 APP/PS1 mice were divided into model group （given Physiological saline）， low and high-dose modified Shuyuwan （14，64 g·kg^-1）group， and donepezil group （1 mg·kg^-1） and 10 wild mice were set as the blank control group （given Physiological saline）. All of the mice were administered intragastrically for 35 days. The memory and space exploration ability of mice was detected by Morris water maze， the morphology of mouse hippocampal neurons were observed by Nissl staining. The deposition of <italic>β </italic>amyloid 1-42（A<italic>β</italic>_1-42） in mouse hippocampus was detected by immunohistochemistry， and the expression of ionized calcium-binding adapter molecule 1（Iba1）， a marker of hippocampal microglia （MG） and Nitric oxide synthase（iNOS）， a marker of actived MG， were detected by immunofluorescence. The protein expression of NLR family pyrin domain containing 3（Nlrp3）， Apoptosis-associated speck-like protein containing a Caspase-recruitment domain （ASC）， cysteine protease-1（Caspase-1）and interleukin-1 beta （IL-1<italic>β</italic>） were detected by Western blot， and the expression of IL-1<italic>β</italic>， tumor necrosis factor-<italic>α</italic>（TNF-<italic>α</italic>）and interleukin-18 （IL-18） mRNA were detected by Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）. Result:Compared with the blank control group， the memory and space exploration ability of the model group were significantly reduced （<italic>P</italic><0.05）， the number of hippocampal neurons decreased， the deposition of A<italic>β</italic>_1-42 increased， the markers of actived MG Iba1，iNOS increased， the protein expression of Nlrp3， ASC， Caspase-1， IL-1<italic>β</italic> increased significantly （<italic>P</italic><0.05）， and the mRNA expression of IL-1<italic>β</italic>， IL-18， and TNF-<italic>α</italic> increased significantly （<italic>P</italic><0.05）. Compared with model group， the Chinese medicine group can improve the APP/PS1 mice's space exploration ability and memory ability （<italic>P</italic><0.05）， increase the number of hippocampal neurons， reduce A<italic>β</italic>_1-42 deposition， reduce the activation of MG， and reduce the protein expression of Nlrp3， ASC， Caspase-1 and IL-1<italic>β</italic> （<italic>P</italic><0.05）， and reduced the expression of IL-1<italic>β</italic> mRNA （<italic>P</italic><0.05）. Conclusion:Modified Shuyuwan can reduce the expression of IL-1<italic>β</italic> and other inflammatory factors in the hippocampus of APP/PS1 mice by inhibiting the Nlrp3/ASC/Caspase-1 pathway， and relieve nerve inflammation and pathological injury of AD.

Effect of Suanzaoren Tang on Learning-memory Function and NLRP3 Pathway in Sleep-deprived Rats / 中国实验方剂学杂志

Objective:To explore the mechanism of Suanzaoren Tang in improving learning-memory of sleep-deprived rats based on Nod-like receptor 3 （NLRP3） inflammatome pathway. Method:The rats were randomly divided into normal control group， model group， Eszolam group（5.4×10^-4 g·kg^-1·d^-1）， low-dose Suanzaoren Tang group（4.59 g·kg^-1·d^-1）and high-dose Suanzaoren Tang group （18.36 g·kg^-1·d^-1）. In addition to normal control group， other groups were used to constructed sleep-deprived model， which was concurrent with 30-day continuous drug administration. Water maze was used to evaluate the learning-memory function of rats； The mRNA and protein expressions of NLRP3， apoptosis-related speckle proteins （ASC）， aspartic acid-specific cysteine protease-1 （Caspase-1）， interleukin-1（IL-1） and IL-18 in the hippocampus of rats were detected by Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） and Western blot. Result:Compared with control group， the incubation period of the platform， the total distance of swimming and the duration of first reaching the platform in model group were significantly increased （<italic>P</italic><0.01）， while the number of platform crossings and the target quadrant time were decreased （<italic>P</italic><0.01）. Compared with the model group， the incubation period， total swimming distance and the duration of first reaching the platform in low-dose Suanzaoren Tang group and high-dose Suanzaoren Tang group were decreased to different degrees （<italic>P</italic><0.05，<italic>P</italic><0.01）， while the number of platform crossings and the target quadrant time were increased significantly （<italic>P</italic><0.05，<italic>P</italic><0.01），but with no significant change in estazolam group. Compared with normal control group， mRNA and protein expressions of NLRP3， ASC， Caspase-1， IL-1<italic>β</italic>， IL-18 in the hippocampus of the model group were significantly increased （<italic>P</italic><0.05，<italic>P</italic><0.01）. Compared with model group， mRNA and protein expressions of NLRP3， ASC， Caspase-1， IL-1<italic>β</italic> and IL-18 in the hippocampus of the rats in low-dose Suanzaoren Tang group and high-dose Suanzaoren Tang group were all decreased to different degrees （<italic>P</italic><0.05）. The mRNA and protein expressions of NLRP3， ASC， Caspase-1， IL-1<italic>β</italic> and IL-18 in the hippocampus of Suanzaoren group also decreased， but with no significant change. Conclusion:Suanzaoren Tang can improve the learning-memory function of sleep-deprived rats， and its mechanism is related to the inhibition of NLRP3 inflammatome pathway in hippocampus and the alleviation of neuroinflammation.

Effect of Modified Buwangsan on Expression of Related Molecules in NLRP3 Inflammatory Signaling Pathway in Hippocampus of AD Rats / 中国实验方剂学杂志

Objective::To investigate the effects of modified Buwangsan on the learning and memory ability of Alzheimer's disease (AD) model rats and the expression of NOD-like receptor 3 (NLRP3), cysteine-containing aspartate-specific proteases 1 (Caspase-1) and interleukin-1 beta (IL-1β) in NLRP3 inflammatory pathway in hippocampus of AD model rats, and exploring the underlying mechanism of modified Buwangsan. Method::The 52 eligible rats were randomly divided into sham control group, AD model group, low-dose modified Buwangsan group (1.5 g·kg-1) and high-dose modified Buwangsan group (3 g·kg-1). AD mouse model was established by bilateral hippocampus injection of Aβ1-425 μL (2 g·L-1). The rats in low-dose and high-dose modified Buwangsan group received low and high dose modified Buwangsan respectively within the next 4 weeks, once daily. The learning and memory ability was tested by Morris water maze. The expression of NLRP3, Caspase-1 and IL-1β mRNA was tested by quantitative PCR(Real-time PCR) and Western blot. Result::As compared with the sham group, the learning and memory ability of the rats were significantly impaired (P<0.05). Compared with AD model group, the learning and memory ability and the expression levels of NLRP3, Caspase-1, and IL-1β mRNA and protein were all no statistical differences in low-dose modified Buwangsan group, while the learning and memory ability of the rats were significantly improved and the expression of NLRP3, Caspase-1 and IL-1β mRNA in hippocampus of rats was significantly decreased in high-dose modified Buwangsan group (P<0.05). Conclusion::High-dose modified Buwangsan could attenuate neuroinflammation in the hippocampus of AD mouse model via inhibiting the expression of NLRP3, Caspase-1 and IL-1β, which may be the mechanisms of modified Buwangsan could be used to ameliorate the learning and memory ability of AD mouse model.