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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 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.
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OBJECTIVE To study the effect of Longsheng zhi capsule on lear ning and memory ability of vascular dementia model rats and explore its mechanism based on metabonomics. METHODS Totally 90 SD rats were randomly divided into sham operation group ，model group ，Dihydroergotoxin mesylate tablet group （positive control ，0.54 mg/kg），Longshengzhi capsule high-dose，medium-dose and low-dose groups （2.16，1.08 and 0.54 g/kg），with 15 rats in each group. In addition to sham operation group （only threading without ligation ），the vascular dementia model was prepared by permanent ligation of bilateral common carotid arteries in each group ，which was administered by gavage for 28 d. Morris water maze test was used to determine the learning and memory ability of rats ；hematoxylin eosin （HE）staining was used to observe the histopathological changes of hippocampus；the serum levels of superoxide dismutase （SOD），malondialdehyde（MDA）and glutathione peroxidase （GSH-Px） were detected ；the serum metabolic map was analyzed by ultra high performance liquid chromatography quadrupole time of flight tandem mass spectrometry （UPLC-Q-TOF/MS），the specific metabolites were screened by multivariate statistical analysis ，and the metabolic pathway was enriched and analyzed. RESULTS Morris water maze test showed that compared with model group ，the escape latency of rats in each administration group was significantly shortened ，the number of crossing the platform was significantly increased ，and the residence time in the target quadrant was significantly prolonged （P＜0.01 or P＜ 0.05）. The results of serum biochemical indexes showed that compared with model group ， the s erum level of SOD increased significantly in Dihydroergotoxine mesylate tablet group and Longshengzhi capsule high-dose group ，the serum level of GSH-Px increased significantly while the MDA level decreased significantly in each administration group （P＜0.05 or P＜0.01）. HE staining showed that Longshengzhi capsule could improve the histopathological damage of hippocampus in vascular dementia model rats. A total of 14 differential metabolites were screened and identified by UPLC-Q/TOF-MS and orthogonal partial least squares discriminant analysis model （VIP＞1 and P＜0.05）. The results of metabolic pathway enrichment analysis showed that the metabolic pathways involved in vascular dementia in rats mainly included vitamin B 6 metabolism，fatty acid metabolism and steroid hormone biosynthesis. CONCLUSIONS Longshengzhi capsule can improve the learning and memory ability of rats caused by vascular dementia. Its effect may be related to improving the oxidative stress injury caused by lipid accumulation in the process of vascular dementia. The metabolic pathways involved mainly include vitamin B 6 metabolism，fatty acid metabolism and steroid hormone biosynthesis.
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ObjectiveTo study the effect of Longshengzhi capsule （LSZC） on high fat diet （HFD）-induced atherosclerosis （AS） in apolipoprotein E knockout （ApoE-/-） mice. MethodApoE-/- mice were fed with HFD for 8 weeks to induce AS. Then the mice were randomized into model group， simvastatin group （4 mg·kg-1）， high-dose LSZC group （1.6 g·kg-1）， medium-dose LSZC group （0.8 g·kg-1）， and low-dose LSZC group （0.4 g·kg-1）. C57BL/6J Mice with normal diet were used as the blank control. After 10 weeks， serum levels of triglyceride （TG）， total cholesterol （TC）， high-density lipoprotein cholesterol （HDL-C）， low-density lipoprotein cholesterol （LDL-C）， malondialdehyde （MDA）， superoxide dismutase （SOD）， interleukin-1β （IL-1β）， and interleukin-6 （IL-6） were detected. Hematoxylin-eosin （HE） and oil red O were used to detect aortic plaque in each group. The levels of CD34 and F4/80 in aorta were determined by immunohistochemistry （IHC）. ResultCompared with the blank control， the model group demonstrated obvious aortic plaque， a large amount of lipid accumulation， serious damage of aortic intima， increase in serum levels of TC， TG， LDL-C， HDL-C， MDA， IL-1β， and IL-6 （P<0.01）， decrease in SOD level （P<0.01）， and rise of the expression of CD34 and F4/80 （P<0.01）. Compared with the model group， LSZC of the three doses all decreased the serum levels of TG and LDL-C （P<0.05）， and the levels of IL-1β and IL-6 （P<0.05， P<0.01）， and the high-dose and medium-dose LSZC improved SOD level， decreased MDA content （P<0.05， P<0.01）， and reduced the expression of the CD34 and F4/80 in blood vessels （P<0.05， P<0.01）. ConclusionLSZC has certain intervention effect on the formation of aortic plaque in atherosclerosis ApoE-/- mice. The mechanism is that it reduces the levels of serum TG and LDL-C to lower blood lipid， decreases MDA level and improves SOD activity to inhibit lipid peroxidation， lowers the levels of IL-1β and IL-6 and down-regulates the expression of CD34 and F4/80 to protect blood vessels from inflammatory damage.
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Objective:To investigate the effect of Longshengzhi capsule (LSZC) on behavior, serum biochemical index, nuclear transcription factor-κB(NF-κB) and cysteine aspartate-specific protease-3(Caspase-3) in stroke rats. Method:Ischaemic stroke was induced by the permanent middle cerebral artery occlusion (pMCAO). The male sprague-dawley rats with successful pMCAO modeling were randomly divided into five groups:model (pMCAO) group, nimodipine (12 mg·kg-1) group and LSZC groups (2.4, 1.2, 0.6 g·kg-1). A sham group also set in the study. The intragastric administration lasted for 14 days. The rats were tested for neurological deficits, grip strength, rotation tolerance. An open field test was also performed to evaluate the behavior. The pathology of the brain was studied by 2,3,5-triphenyltetrazolium chloride (TTC) staining, and hematoxylin and eosin (HE) staining. In addition, the levels of malondialdehyde (MDA), nitric oxide (NO) and superoxide dismutase (SOD) in rat serum were tested in the experiment. Immunohistochemistry (ICH) and Western blot were used to analyze the expressions of Caspase-3 and NF-κB. Result:Compared the model group, LSZC significantly improved the grip strength, rotation tolerance and spontaneous activity. LSZC markedly reduced infarct volume, water content of brain, neurological scores, body weight decline in MCAO stroke rats (PPPPκB. Conclusion:LSZC treatment can improve neurological deficit function and protect the brain of stroke rats. This effect may be realized by down-regulating the expressions of NF-κB and Caspase-3.