Protective effect and mechanism of Longshengzhi capsules on cerebral ischemia-reperfusion injury in rats
OBJECTIVE To explore the protective effect and mechanism of Longshengzhi capsules on cerebral ischemia-reperfusion injury in rats.METHODS The model of middle cerebral artery occlusion(MCAO)was established by using the improved thread occlusion method.The experiment was divided into six groups:sham surgery group(only separating blood vessels without inserting thread plugs,given the same volume of normal saline),model group(modeling,given the same volume of normal saline),nimodipine group(positive control,modeling,dose of 20 mg/kg),and low-dose,medium-dose,and high-dose groups of Longshengzhi capsules(modeling,doses of 0.72,1.44 and 2.88 g/kg,respectively),with 10 mice in each group.Each group was given corresponding medication solution/normal saline by gavage,once a day,for 7 consecutive days.One hour after the last administration,the Zea Longa scoring method was used to score the neurological deficits in each group of rats,and the ABC enzyme-linked immunosorbent assay was used to detect the serum levels of tumor necrosis factor-α(TNF-α)and interleukin-6(IL-6)in rats;TTC staining was used to observe the volume of cerebral infarction in rats and calculate the cerebral infarction volume ratio.Hematoxylin eosin staining was used to observe the pathological changes in the brain tissue of rats.Immunohistochemical staining was used to detect the positive expression of NLRP3 protein in the brain tissue of rats.Real-time fluorescence quantitative PCR was used to detect mRNA relative expressions of Toll-like receptor 4(TLR4)and nuclear factor-κB(NF-κB)in the brain tissue of rats.Western blot assay was adopted to detect the relative expressions of TLR4,NLRP3 and phosphorylated NF-κB(p-NF-κB)protein in the brain tissue of rats and its intracellular NF-κB protein.RESULTS Compared with the sham surgery group,the neural dysfunction score,serum levels of TNF-α and IL-6,cerebral infarction volume ratio,relative expression levels of NF-κB and TLR4 mRNA,as well as protein relative expressions of TLR4,NLRP3 and p-NF-κB in the brain tissue,and relative protein expression of intracellular NF-κB were increased significantly in the model group(P＜0.01);the enlarged gap and significant edema were observed in cortical nerve cells of dysfunction score,serum levels of TNF-α and IL-6,cerebral infarction volume ratio,relative expression levels of NF-κB and TLR4 mRNA,as well as protein relative expressions of TLR4,NLRP3 and p-NF-κB in the brain tissue,and relative protein expression of intracellular NF-κB were increased significantly in the model group(P＜0.01);the enlarged gap and significant edema were observed in cortical nerve cells of dysfunction score,serum levels of TNF-α and IL-6,cerebral infarction volume ratio,relative expression levels of NF-κB and TLR4 mRNA,as well as protein relative expressions of TLR4,NLRP3 and p-NF-κB in the brain tissue,and relative protein expression of intracellular NF-κB were increased significantly in the model group(P＜0.01);the enlarged gap and significant edema were observed in cortical nerve cells of brain tissue in rats,with a large amount of inflammatory cell infiltration;the positive expression of NLRP3 protein in brain tissue of rats obviously increased.Compared with the model group,the levels of the above indicators in the medium-dose and high-dose groups of Longshengzhi capsules,as well as the Nimodipine group,were reversed to varying degrees,and most differences were statistically significant(P＜0.05 or P＜0.01);the pathological morphology observation showed a significant improvement,and the positive expression of NLRP3 protein in the brain tissue of rats was obviously reduced.CONCLUSIONS Longshengzhi capsules may inhibit TLR4/NF-κB/NLRP3 signaling pathway and neuroinflammatory response,thereby achieving a protective effect against cerebral ischemia-reperfusion injury in rats.
万方数据
维普
