[Research status of insect infestation of Chinese medicinal materials during storage].

During the storage process, Chinese medicinal materials are susceptible to insect infestation due to their own nature and external storage factors. Infestation by insects can have varying impacts on the materials. In mild cases, it affects the appearance and reduces consumer purchasing power, while in severe cases, it affects the quality, reduces medicinal value, and introduces impurities such as insect bodies, excrement, and secretions, resulting in significant contamination of the medicinal materials. This study reviewed the rele-vant factors influencing insect infestation in Chinese medicinal materials and the compositional changes that occur after infestation and summarized maintenance measures for preventing insect infestation. Additionally, it provided an overview of detection techniques applicable to identifying insect infestation during the storage of Chinese medicinal materials. During the storage process, insect infestation is the result of the combined effects of biological factors(source, species, and population density of insects), intrinsic factors(moisture, chemical composition, and metabolism), and environmental factors(temperature, relative humidity, and oxygen content). After infestation, there are significant changes in the content of constituents in the medicinal materials. By implementing strict pre-storage inspections, regular maintenance after storage, and appropriate storage and maintenance methods, the occurrence of insect infestation can be reduced, and the preservation rate of Chinese medicinal materials can be improved. The storage and maintenance of Chinese medicinal materials are critical for ensuring their quality. Through scientifically standardized storage and strict adherence to operational management standards, the risk of insect infestation can be minimized, thus guaranteeing the quality of Chinese medicinal materials.

Saponins and their derivatives: Potential candidates to alleviate anthracycline-induced cardiotoxicity and multidrug resistance.

Anthracyclines (ANTs) continue to play an irreplaceable role in oncology treatment. However, the clinical application of ANTs has been limited. In the first place, ANTs can cause dose-dependent cardiotoxicity such as arrhythmia, cardiomyopathy, and congestive heart failure. In the second place, the development of multidrug resistance (MDR) leads to their chemotherapeutic failure. Oncology cardiologists are urgently searching for agents that can both protect the heart and reverse MDR without compromising the antitumor effects of ANTs. Based on in vivo and in vitro data, we found that natural compounds, including saponins, may be active agents for other both natural and chemical compounds in the inhibition of anthracycline-induced cardiotoxicity (AIC) and the reversal of MDR. In this review, we summarize the work of previous researchers, describe the mechanisms of AIC and MDR, and focus on revealing the pharmacological effects and potential molecular targets of saponins and their derivatives in the inhibition of AIC and the reversal of MDR, aiming to encourage future research and clinical trials.

Characterization of chemical composition variations in raw and processed Codonopsis Radix by integrating metabolomics and glycomics based on multiple chromatography-mass spectrometry technology.

Codonopsis Radix, a popular food homology medicine, is widely used in clinical traditional Chinese medicine and food supplement, raw products and three types of processed products are the main forms of decoction pieces in China. However, there is no scientific basis for comprehensive chemical characterization of raw and three types of processed products. Herein, we investigated qualitatively and quantificationally secondary and primary metabolites in raw Codonopsis Radix and three types of processed products by metabolomics and glycomics employing multiple chromatography-mass spectrometry technology combined with chemometric analysis further to look for differential compounds and propose the processing-induced chemical mechanisms. The results indicated that Codonopsis Radix became dark-colored and the smell of burnt incense odor was observed after processing. The principal component analysis demonstrated that secondary metabolome and glycome were significantly altered between raw and processed products, and 36 differential secondary metabolites and 11 differential primary metabolites were finally screened through orthogonal partial least-squares-discriminant analysis. The main types of compounds are alkaloids, terpenoids, glycosides, amino acids, monosaccharides, oligosaccharides, and furfural derivatives. Meanwhile, Chemical mechanisms could be involved, including oxidation, glycosidic hydrolysis, esterification, dehydration, and Maillard reaction. This work supplies a chemical basis for the application of various types of Codonopsis Radix decoction pieces.

TREM-1-targeting LP17 attenuates cerebral ischemia-induced neuronal injury by inhibiting oxidative stress and pyroptosis.

Stroke ranks as the second leading cause of disability and death globally. Trigger receptors expressed on myeloid cells (TREM) -1 are responsible for the activation of the innate immune response and also play a critical role in inflammation. In this study, we reported the contribution of TREM-1 after ischemic damage in a rat middle cerebral artery occlusion (MCAO) model. This study also demonstrated that TREM-1 expression was upregulated following cerebral infarction in rats. TREM-1 inhibition was determined using its selective inhibitor, LP17, which indicated a neuroprotective effect on cerebral infarction damage. The findings revealed that inhibition of TREM-1 by administering LP17 improved cerebral damage and decreased ischemic areas and brain water contents. Moreover, LP17 decreased MCAO-induced microglial activation and neurodegeneration, evidenced by a reduction in the expression of microglial Iba-1 and FJ-B positive cells, and reversed neuronal loss. Besides, the contribution of LP17 to ischemic neuronal damage may be associated with a decrease in the production of pro-inflammatory cytokines, and enhanced production of anti-inflammatory cytokine IL-10. Both in vivo and in vitro studies showed that inhibiting TREM-1 attenuated ROS accumulation, lipid per-oxidation (LPO) contents such as malondialdehyde (MDA) and enhanced the superoxide dismutase (SOD) activity after ischemia. Inhibiting TREM-1 alleviated inflammation and pyroptosis found in MCAO rats. This was achieved through the inhibition of the levels of NLRP3, caspase-1, ASC (an apoptosis-associated speck-like protein containing a CARD) and gasdermin D. These results confirmed that inhibiting TREM-1 protects against ischemia-induced neuronal damage and alleviates microglial mediated neuro-inflammation by reducing oxidative stress and pyroptosis. Therefore, blocking TREM-1 expression provides an effective intervention for improving ischemic stroke.

[Chemical constituents of Angelica nitida roots].

OBJECTIVE: To investigate the chemical constituents in the roots of Angelica nitida. METHODS: The chemical constituents were isolated by silica gel,their structures were identified by spectral analysis. RESULTS: Nine compounds were isolated and identified as isoimperatorin(I), imperatorin(II), cnidilin(III), beta-sitosterol(IV), isopimpinellin(V), phellopterin(VI), neobyakangelicol(VII), (3S)-2,2-dimethyl-3,5-dihydroxy-8-hydroxylmethyl-3,4-dihydro-2H,6H-benzo[1,2-b: 5,4-b'] dipyran-6-one(VIll) and byakangelicin (IX). CONCLUSION: All the compounds are isolated from the plant for the first time.

[Comparative study on HPLC fingerprint of ordinary powder and ultrafine powder for Gardenia jasminoides f. longicarpa].

OBJECTIVE: To establish an HPLC fingerprint of Gardenia jasminoides f. longicarpa and compare the differences between its ordinary powder and ultrafine powder. METHODS: The analysis was carried out on a Kromasil C18 (250 mm x 4.6 mm, 5 microm) column with gradient elution of acetonitrile-0.4% phosphoric acid at the flow rate of 1.0 ml/min. The wavelength was 240 nm during 0 - 40 min and 440 nm during 40 - 80 min. RESULTS: HPLC fingerprint of Gardenia jasminoides f. longicarpa was established, 23 common peaks were identified,and the similarity of 10 samples was greater than 0.9. Ultrafine grinding did not change the types and number of chemical compositions, but it obviously increased the content of main chemical compositions. CONCLUSION: The HPLC fingerprint is accurate, reliable and repeatable, which can be used for quality control of Gardenia jasminoides f. longicarpa. Ultrafine grinding can stimulate the release of chemical components of Gardenia jasminoides f. longicarpa.

Alpha-Asarone modulates kynurenine disposal in muscle and mediates resilience to stress-induced depression via PGC-1α induction.

INTRODUCTION: Kynurenine (KYN) accumulation in periphery induces brain injury, responsible for depression. α-Asarone is a simple phenylpropanoids that exerts beneficial effects on central nervous system. However, the effect of α-asarone on periphery is unexplored. AIMS: Here, we investigated its protective role against depression from the aspect of KYN metabolism in skeletal muscle. METHODS: The antidepressant effects of α-asarone were evaluated in chronic mild stress (CMS) and muscle-specific PGC-1α-deficient mice. The effects of KYN metabolism were determined in mice and C2C12 myoblasts. RESULTS: α-Asarone exerted antidepressant effects in CMS and KYN-challenged mice via modulating KYN metabolism. In myoblasts, α-asarone regulated PGC-1α induction via cAMP/CREB signaling and upregulated KYN aminotransferases (KATs) to increase KYN clearance in a manner dependent on PGC-1α. KAT function is coupled with malate-aspartate shuttle (MAS), while α-asarone combated oxidative stress to protect MAS and mitochondrial integrity by raising the NAD+ /NADH ratio, ensuring effective KYN disposal. In support, the antidepressant effect of α-asarone was diminished by muscle-specific PGC-1α deficient mice subjected to KYN challenge. CONCLUSION: KATs coupled with MAS to clear KYN in muscle. α-Asarone increased PGC-1α induction and promoted KYN disposal in muscle, suggesting that protection of mitochondria is a way for pharmacological intervention to depression.

Metagenomic next-generation sequencing in the diagnosis of neurocysticercosis: A case report.

BACKGROUND: The clinical symptoms and imaging manifestations of neurocysticercosis (NCC) are very different, and the difficulty and delay of clinical diagnoses may lead to an increase in mortality and disability. Rapid and accurate pathogen identification is important for the treatment of these patients. Metagenomic next-generation sequencing (mNGS) is a powerful tool to identify pathogens, especially in infections that are difficult to identify by conventional methods. CASE SUMMARY: A 43-year-old male patient was admitted due to a recurrent headache for a few months. Imaging examinations showed hydrocephalus and cystic lesions, which were considered to be a central nervous system infection, but no etiology was found by routine examination. mNGS of the cerebrospinal fluid revealed high Taenia solium reads, and the positive results of a cysticercosis antibody test confirmed the infection. Combined with the patient's clinical manifestations, the etiological evidence, and the imaging manifestation, the patient was finally diagnosed with NCC and he was prescribed dexamethasone, albendazole, neurotrophic drugs, and intracranial pressure reduction therapy. The headaches disappeared after anti-parasite treatment, and no associated symptoms recurred prior to the three- and six-month follow-up. CONCLUSION: As an accurate and sensitivity detection method, mNGS can be a reliable approach for the diagnosis of NCC.

Crucial Role of the Ca2+/CN Signaling Pathway in the Antifungal Activity of Seselin against Botrytis cinerea.

Botrytis cinerea causes gray mold in many fruit and vegetable crops. We previously found that Seselin (SL) displayed antifungal activity against B. cinerea (EC50 = 6.1 µg·mL-1), and this study investigated the effects of Ca2+ and the Ca2+/CN signaling pathway on its antifungal activity against B. cinerea. The results indicated that exogenous Ca2+, Cyclosporine A, and Verapamil reduced the sensitivity of SL against B. cinerea; SL significantly reduced the intracellular Ca2+ concentration in the hyphae; the sensitivity of strains ΔbcCCH1 and ΔbcMID1 to SL were significantly increased; and the expressions of CCH1, MID1, CNA, PMC1, and PMR1 genes of the Ca2+/CN signaling pathway were significantly downregulated by SL treatment. Hence, SL is a potential compound for developing fungicides against B. cinerea. SL dramatically reduces intracellular Ca2+ concentration and disturbs Ca2+ homeostasis, leading to cell death. The Ca2+/CN signaling pathway plays an important role in the antifungal activity of SL against B. cinerea.

Crucial role of Ca2+ /CN signalling pathway in the antifungal activity of disenecioyl-cis-khellactone against Botrytis cinerea.

BACKGROUND: Botrytis cinerea causes grey mould and is one of the most destructive fungal pathogens affecting important fruit and vegetable crops. In preliminary studies, we found that disenecioyl-cis-khellactone (DK) had strong antifungal activity against several fungi species including B. cinerea [half maximal effective concentration (EC50 ) = 11.0 µg mL-1 ]. In this study, we aimed to further evaluate the antifungal activity of DK against B. cinerea and determine the role of calcium ion/calcineurin (Ca2+ /CN) signalling pathway on its antifungal effect. RESULTS: DK was effective against B. cinerea in both in vitro and in vivo assays. Exogenous Ca2+ reduced the antifungal activity of DK. The combination of DK and cyclosporine A (CsA) did not exhibit an additive effect against B. cinerea. In contrast to CsA, DK reduced the intracellular Ca2+ concentration in B. cinerea. DK bound to calcineurin A (cnA) and up-regulated the expression of PMC1 and PMR1 genes. Moreover, DK sensitivity of △bccnA significantly decreased compared with that of Bc05.10 strain. CONCLUSION: DK is a promising lead compound for developing fungicides against B. cinerea. The Ca2+ /CN signalling pathway plays a crucial role in the DK antifungal activity, and cnA is one of the targets of DK against B. cinerea. DK directly reacts with cnA, which up-regulates the transcription of Ca2+ /CN-dependent target genes PMC1 and PMR1, decreasing the intracellular Ca2+ concentration and disturbing the intracellular Ca2+ balance, leading to cell death. © 2022 Society of Chemical Industry.

Ginkgo biloba exocarp extracts inhibit S. aureus and MRSA by disrupting biofilms and affecting gene expression.

ETHNOPHARMACOLOGICAL RELEVANCE: Ginkgo biloba L. fruit, also known as Bai Guo, Ya Jiao Zi (in pinyin Chinese), and ginkgo nut (in English), has been used for many years as an important material in Chinese traditional medicine to treat coughs and asthma and as a disinfectant, as described in the Compendium of Materia Medica (Ben Cao Gang Mu, pinyin in Chinese), an old herbal book. Ginkgo nuts are used to treat phlegm-associated asthma, astringent gasp, frequent urination, gonorrhoea and turgidity; consumed raw to reduce phlegm and treat hangovers; and used as a disinfectant and insecticide. A similar record was also found in Sheng Nong's herbal classic (Shen Nong Ben Cao Jing, pinyin in Chinese). Recent research has shown that Ginkgo biloba L. exocarp extract (GBEE) can unblock blood vessels and improve brain function and exhibits antitumour and antibacterial activities. AIM OF STUDY: To investigate the inhibitory effect of Ginkgo biloba L. exocarp extract (GBEE) on methicillin-resistant S. aureus (MRSA) biofilms and assess its associated molecular mechanism. MATERIALS AND METHODS: The antibacterial effects of GBEE on S. aureus and MRSA were determined using the broth microdilution method. The growth curves of bacteria treated with or without GBEE were generated by measuring the CFU (colony forming unit) of cultures at different time points. The effects of GBEE on bacterial biofilm formation and mature biofilm disruption were determined by crystal violet staining. Quantitative polymerase chain reaction (qPCR) was used to measure the effects of GBEE on the gene expression profiles of MRSA biofilm-related factors at 6, 8, 12, 16 and 24 h. RESULTS: The minimum inhibitory concentration (MIC) of GBEE on S. aureus and MRSA was 4 µg/mL, and the minimum bactericidal concentration (MBC) was 8 µg/ml. Moreover, GBEE (4-12 µg/mL) inhibited S. aureus and MRSA biofilm formation in a dose-dependent manner. Interestingly, GBEE also destroyed mature biofilms of S. aureus and MRSA at 12 µg/ml. The expression of the MRSA biofilm-associated factor icaA and sarA were downregulated after 6 h of treatment with GBEE, while sigB was downregulated after 12 h. MeanwhileMeanwhile, icaR was upregulated at 12 h. In addition, GBEE also downregulated the virulence gene hld and inhibited the synthesis of staphyloxanthin. CONCLUSIONS: GBEE has excellent antibacterial effects against S. aureus and MRSA and inhibits their biofilm-forming ability by altering related gene expression.

[Studies on the chemical constituents in roots of Angelica tianmuensis and A. megaphylla].

OBJECTIVE: To study the chemical constituents in roots of Angelica tianmuensis and A. megaphylla. METHODS: Compounds were isolated by column chromatography with silica gel, their structures were identified by spectral analysis. RESULTS: Another three compounds (ligustilone, 5-methoxyhamaudol, cimifugin) were obtained from the roots of Angelica tianmuensis, and another two compounds (ligustilone, angelol) were obtained from the roots of A. megaphylla. CONCLUSION: All the compounds are isolated in these two plants for the first time, and ligustilone is first found from Angelica L..

Memantine ameliorates tau protein deposition and secondary damage in the ipsilateral thalamus and sensory decline following focal cortical infarction in rats.

Previous studies have reported that memantine presents evidence of therapeutic benefits in several animal models of ischemic stroke and neurodegenerative diseases. However, the effect of memantine on secondary damage in the ipsilateral thalamus after focal cortical infarction remains undefined. Present study investigated whether memantine has a protective effect on secondary damage in the ipsilateral thalamus after focal cerebral infarction in rats. At 24 h after distal middle cerebral artery occlusion (MCAO), rats in the memantine and vehicle groups were intraperitoneal injected with memantine and isopycnic vehicle, respectively, was once daily administered for consecutive 7 days. Infarct size was evaluated through Nissl staining and sensory decline determined using adhesive removal test. Secondary thalamic damage was assessed using Nissl staining and immunofluorescence 8 days after MCAO. Immunoboltting was used to identify tau and apoptosis-associated proteins in the ipsilateral thalamus after MCAO. Results revealed that memantine ameliorated sensory decline compared to the vehicle controls. Subsequently, tau phosphorylated at threonine 231 (p-tau-231), glycogen synthase kinase3ßpY216 (GSK3ßpY216) and protein phosphatase 2A (PP2ApY307) were reduced by memantine, causing greater reduction in neuronal loss and inhibition of reactive astrogliosis in the ipsilateral ventroposterior thalamic nucleus (VPN) compared with the vehicle groups. In addition, increase in secondary damage-induced TUNEL-positive cells was blunted by memantine, as demonstrated by the significant reduction in expression of apoptosis-associated proteins. Our results suggest that memantine has a neuro-protective effect on secondary damage in the ipsilateral thalamus following MCAO by inhibiting the activity of GSK3ßpY216/PP2ApY307 and down regulating the levels of p-tau-231 protein.

Clinical significance of tumor cells in the peripheral blood of patients with esophageal squamous cell carcinoma.

Circulating tumor cells (CTCs) are suspected of predicting the prognosis of malignant tumor, but there are few relevant reports specific to esophageal squamous cell carcinoma (ESCC). This study investigated the clinical significance of CTCs in patients with ESCC.Sixty patients with ESCC were enrolled, from whom CTCs had been tested by our team previously. Peripheral blood samples were obtained from these patients before treatment; and CTCs were assayed by isolation by size of epithelial tumor cells (ISET). Associations between the presence of CTCs and patients' clinicopathological parameters and clinical outcomes were analyzed.CTCs were detected in 20 patients (33.3%), who experienced significantly shorter progression-free survival (PFS) than did the CTC-negative patients. Overall, PFS was negatively associated with the number of CTCs. Multivariate analyses showed that a CTC count >2 was a strong independent prognostic indicator of tumor recurrence (hazard ratio [HR] 5.63; 95% confidence interval [CI] 1.77-17.89; P = .003). In the subgroup of 50 patients who underwent R0 resection and postoperative adjuvant radiotherapy or chemotherapy, CTC was a strong, independent, and prognostic indicator of tumor recurrence (HR 10.70; 95% CI, 1.40-81.91; P = .022). The number of CTCs correlated with the T stage (r = 0.26, P = .043) but not with the N or M stage. For subgroups in stages II or I-IIIB or T3 or T3 + T4, the PFS of patients with CTCs > 1 or > 2 was significantly shorter than that of the patients with CTCs ≤ 1 or CTCs ≤ 2. In the stage III or T3 + T4 groups, the PFS of patients with CTCs > 0 was significantly shorter than that of patients with CTC = 0.This is the first study to report that the CTC detected by ISET is an independent and prognostic indicator of patients' outcome in ESCC. Consideration of CTCs may improve the accuracy of preoperative staging in ESCC.

Acute toxicity, bioconcentration, elimination and antioxidant effects of fluralaner in zebrafish, Danio rerio.

Fluralaner is a novel isoxazoline insecticide which shows high insecticidal activity against parasitic, sanitary and agricultural pests, but there is little information about the effect of fluralaner on non-target organisms. This study reports the acute toxicity, bioconcentration, elimination and antioxidant response of fluralaner in zebrafish. All LC50 values of fluralaner to zebrafish were higher than 10 mg L-1 at 24, 48, 72 and 96 h. To study the bioconcentration and elimination, the zebrafish were exposed to sub-lethal concentrations of fluralaner (2.00 and 0.20 mg L-1) for 15 d and then held 6 d in clean water. The results showed medium BCF of fluralaner with values of 12.06 (48 h) and 21.34 (144 h) after exposure to 2.00 and 0.20 mg L-1 fluralaner, respectively. In the elimination process, a concentration of only 0.113 mg kg-1 was found in zebrafish on the 6th day after removal to clean water. After exposure in 2.00 mg L-1 fluralaner, the enzyme activities of SOD, CAT, and GST, GSH-PX, CarE and content of MDA were measured. Only CAT and CarE activities were significantly regulated and the others stayed at a stable level compared to the control group. Meanwhile, transcriptional expression of CYP1C2, CYP1D1, CYP11A were significantly down-regulated at 12 h exposed to 2.00 mg L-1 of fluralaner. Except CYP1D1, others CYPs were up-regulated at different time during exposure periods. Fluralaner and its formulated product (BRAVECTO®) are of low toxicity to zebrafish and are rapidly concentrated in zebrafish and eliminated after exposure in clean water. Antioxidant defense and metabolic systems were involved in the fluralaner-induced toxicity. Among them, the activities of CAT and CarE, and most mRNA expression level of CYPs showed fast response to the sub-lethal concentration of fluralaner, which could be used as a biomarker relevant to the toxicity.

MiR-29c reduces the cisplatin resistance of non-small cell lung cancer cells by negatively regulating the PI3K/Akt pathway.

In previous studies, miR-29s showed tumor suppressor properties against lung cancer, which improved the survival of patients upon the administration of chemotherapy via an unknown mechanism. Here, we investigated the regulatory effects of miR-29s on the cisplatin resistance of NSCLC cells. The expression of miR-29s was assessed in 130 clinical patients and in cisplatin-treated NSCLS cell lines. MiR-29c expression was decreased in 77% of NSCLC patients. Cisplatin treatment increased the expression of miR-29c and decreased the expression of its oncogenic target AKT2 in NSCLC cell lines. A Kaplan-Meier survival analysis indicated that higher miR-29c levels led to a longer disease-free survival. In particular, patients who experienced cancer recurrences after cisplatin chemotherapy exhibited a lower level of miR-29c expression, suggesting that miR-29c activation may contribute to the chemotherapeutic efficiency of cisplatin. The enforced expression of miR-29c enhanced the cisplatin sensitivity of NSCLC cells, while the knocking down of miR-29c led to cisplatin resistance. MiR-29c amplified the therapeutic effects of cisplatin in vivo. Rescue experiments suggested that miR-29c regulates the cisplatin resistance of NSCLS cells by negatively regulating the PI3K/Akt pathway. Overall, our results demonstrated that miR-29c enhances the sensitivity of NSCLC cells to cisplatin by targeting the PI3K/Akt pathway.

Evaluation of antifungal activities and structure-activity relationships of coumarin derivatives.

BACKGROUND: Osthol is a natural coumarin and lead compound that has been developed into commercial fungicides in China. Natural coumarins comprise five major subtypes: simple coumarins, linear furanocoumarins, angular furanocoumarins, linear pyranocoumarins and angular pyranocoumarins. Studies pertaining to the antifungal activities of linear pyranocoumarins are few, and no reports exist for the antifungal activities of angular pyranocoumarins. In order to discover more antifungal natural coumarins, we synthesised a series of simple natural coumarins and isolated several plant-based furanocoumarins and pyranocoumarins using previously described methods. The compounds were biologically evaluated against some plant fungal pathogens. RESULTS: Several of the 35 coumarins evaluated here exhibited strong activities against specific fungal species, including compound 25 (Pd-D-V, a linear pyranocoumarin), compound 26 (libanorin, an angular furanocoumarin) and compound 34 (disenecioyl khellactone, an angular pyranocoumarin). Compound 25 exhibited a high activity against Sclerotinia sclerotiorum (EC50 = 13.2 µg mL-1 ); compound 34 displayed a strong antifungal activity against Botrytis cinerea (EC50 = 11.0 µg mL-1 ). CONCLUSION: This study demonstrates that several natural coumarins (one linear pyranocoumarin and one angular pyranocoumarin in particular) exhibit strong antifungal activities. These results call for further studies, where these coumarins can be examined as potential lead compounds for developing novel antifungal agents. © 2016 Society of Chemical Industry.

[Effect of lidocaine on the impairment of learning and memory function and central cholinergic system after transient global cerebral ischemia in mice].

OBJECTIVE: To evaluate the effects of lidocaine on the impairments of learning and memorial function and central cholinergic system after transient global cerebral ischemia in mice of different apolipoprotein E genotypes. METHODS: Transient global ischemia was induced by bilateral common carotid arteries occlusion (BCCAO) for 17 minutes. Healthy male C57BL/6J wild-type mice (C57 mice) and apolipoprotein E knockout mice (ApoE mice) were randomly divided into six groups: C57 control group (sham operation, neither BCCAO was performed nor pharmacologic intervention was given), C57 ischemia group (BCCAO for 17 minutes was performed and normal saline was given intraperitoneally), C57 lidocaine group (BCCAO for 17 minutes was performed and lidocaine was given intraperitoneally), ApoE control group (the same procedure as that of C57 control group), ApoE ischemia group (the same procedure as that of C57 ischemia group), ApoE lidocaine group (the same procedure as that of C57 lidocaine group). The mice were allowed to recover for 7 days. Morris water maze test were performed from the 8th postoperative day. Mice were tested four times daily for 5 consecutive days. The latency periods were recorded and the percentages of effective search strategies were calculated. On the 12th postoperative day after Morris water maze test, mice were decapitated under anesthesia. The cerebral cortex and hippocampus were removed quickly. The activities of acetylcholinesterase (AChE) and choline acetyltransferase (ChAT) as well as the binding activity of muscarinic receptor (M receptor) were assayed. RESULTS: (1) The latency periods were significantly longer in the ischemia groups than in the corresponding control groups (P<0.05 or 0.01). They were also significantly longer in C57 lidocaine group than in C57 ischemia group [on the 3rd day of test, (74.1+/-32.7)s vs (49.2+/-19.5)s] (P<0.05). However, they were significantly shorter in apoE lidocaine group than in apoE ischemia group [from the 3rd to the 5th days of test, (40.7+/-27.7)s vs (84.7+/-26.8)s, (31.2+/-19.2)s vs (72.1+/-33.0)s, and (28.0+/-22.1)s vs (60.8+/-26.9)s, respectively] (P<0.05 or 0.01). When compared between two strains, they were significantly longer in apoE ischemia group than in C57 ischemia group (P<0.05 or 0.01). However, they were significantly shorter in apoE lidocaine group than in C57 lidocaine group (P<0.01). (2) The percentages of effective search strategies were significantly lower in the ischemia groups than in the corresponding control groups (P<0.01). They were also significantly lower in C57 lidocaine group than in C57 ischemia group [from the 3rd to the 5th days of test, (18.2+/-11.7)% vs (41.7+/-17.7)%, (22.7+/-20.8)% vs (55.6+/-20.8)%, and (29.6+/-27.0)% vs (66.7+/-21.7)%, respectively] (P<0.01). However, they were significantly higher in apoE lidocaine group than in apoE ischemia group [from the 3rd to the 5th days of test, (41.7+/-25.8)% vs (15.6+/-12.9)%, 8.3+/-20.4)% vs (18.8+/-11.6)%, and (66.7+/-30.3)% vs (28.1+/-20.9)%, respectively] (P<0.01). When compared between two strains, they were significantly lower in apoE ischemia group than in C57 ischemia group (P<0.01). However, they were significantly higher in apoE lidocaine group than in C57 lidocaine group (P<0.01). (3) The parameters of central cholinergic system were significantly lower in the ischemia groups than in the corresponding control groups (P<0.05 or 0.01). They were also significantly lower in C57 lidocaine group than in C57 ischemia group [the activities of AChE of cerebral cortex and hippocampus, (0.44+/-0.09) U/mg protein vs (0.57+/-0.08) U/mg protein, and (0.73+/-0.21) U/mg protein vs (1.08+/-0.27) U/mg protein, respectively; the activities of ChAT of hippocampus, (80.60+/-6.55) pmol/mg protein/min vs (93.66+/-11.15) pmol/mg protein/min; and the binding activities of M receptor of cerebral cortex and hippocampus, (6.03+/-0.74) pmol/mg protein vs (7.49+/-0.48) pmol/mg protein, and (7.56+/-0.92) pmol/mg protein vs (10.65+/-3.35) pmol/mg protein, respectively] (P< 0.05 or 0.01). However, they were significantly higher in ApoE lidocaine group than in ApoE ischemia group [the activities of ChAT of cerebral cortex and hippocampus, (66.99+/-7.55) pmol/mg protein/min vs (46.23+/-4.96) pmol/mg protein/min, and (116.46+/-24.05) pmol/mg protein/min vs (92.08+/-16.33) pmol/mg protein/min, respectively] (P<0.05 or 0.01). When compared between two strains, they were significantly higher in ApoE lidocaine group than in C57 lidocaine group (P< 0.05 or 0.01). CONCLUSION: Transient global cerebral ischemia caused significant brain damages in both strains of mice, which were represented by decline of learning and memorial function and damage of the central cholinergic system. Compared with the C57 mice, the ApoE mice had enhanced susceptibility to global cerebral ischemic injury as shown by more severe decline of the learning and memorial function. In the C57 mice, lidocaine significantly worsened the ischemic brain damage. In the ApoE mice, however, lidocaine significantly alleviated the ischemic cerebral results.

[Effects of lidocaine on brain damage induced by transient global cerebral ischemia in mice of different apolipoprotein E genotypes].

OBJECTIVE: To evaluate the effects of lidocaine on changes of neuropathological outcome as well as the learning and memory abilities induced by transient global cerebral ischemia in mice of different apolipoprotein E genotypes. METHODS: Transient global ischemia was induced by bilateral common carotid arteries occlusion (BCCAO) for a period of 17 minutes. Healthy male C57BL/6J wild-type mice (C57 mice) and apolipoprotein E knockout mice (apoE mice) were randomly divided into six groups: C57 control group (n = 15, undergoing sham operation, neither BCCAO was performed nor pharmacologic intervention was given), C57 ischemia group (n = 21, BCCAO for 17 minutes was performed and normal saline was given intraperitoneally), C57 lidocaine group (n = 22, BCCAO for 17 minutes was performed and lidocaine was given intraperitoneally), apoE control group (n = 15, undergoing the same procedure as that of the C57 control group), apoE ischemia group (n = 19, undergoing the same procedure as that of the C57 ischemia group), apoE lidocaine group (n = 16, undergoing the same procedure as that of the C57 lidocaine group). The mice were allowed to recover for 7 days. Twenty-eight mice were randomly selected from these 6 groups for neuropathological studies on the 7 th postoperative day. The percentage of ischemic neurons in the CA1 region of hippocampus was calculated. Morris water maze tasks were performed for the rest mice from the 8 th postoperative day. Mice were tested four times daily for 5 consecutive days. The latency period was recorded and the percentage of effective search strategies were calculated. RESULTS: (1) The percentage of ischemic neurons in the CA1 region of hippocampus was 0.3% +/- 0.1% in the C57 control group, 19.3% +/- 4.5% in the C57 ischemia group, 36.9% +/- 2.5% in the C57 lidocaine group, 0.6% +/- 0.3% in the apoE control group, 65.5% +/- 2.2% in the apoE ischemia group, and 39.4% +/- 6.5% in the apoE lidocaine group, significantly higher in the ischemia and lidocaine groups than in the corresponding control groups (all P < 0.01), significantly higher in the C57 lidocaine and apoE ischemia groups than in the C57 ischemia group (both P < 0.01), however, significantly lower in the apoE lidocaine group than in the apoE ischemia group (both P < 0.01). (2) The latency period decreased significantly along with the test days in all groups except in the apoE ischemia group (P < 0.05 or 0.01), significantly longer in the ischemia and lidocaine groups than in the corresponding control groups (P < 0.05 or 0.01), significantly longer in the C57 lidocaine group than in the C57 ischemia group on the 3 rd day of test [73.1 (22.1-120.1) s vs. 40.2 (28.4-91.1) s], and in the apoE ischemia group than in the C57 ischemia group on the 3 rd and 4 th days of test [88.2 (41.0-120.1) s vs. 40.2 (28.4-91.1) s, and 78.2 (32.9-120.1) s vs. 46.3 (11.6-81.9) s] (P < 0.05 or 0.01), and, however, significantly shorter in the apoE lidocaine group than in the C57 lidocaine group on the 3rd day of test [39.0 (15.5-103.5) s vs. 73.1 (22.1-120.1) s], and in the apoE lidocaine group than in the apoE ischemia group from the 3 rd to the 5 th days of test [39.0 (15.5-103.5) s vs. 88.2 (41.0-120.1) s, 24.9 (11.8-68.0) s vs. 78.2 (32.9-120.1) s, and 29.1 (6.6-57.2) s vs. 66.3 (14.2-97.0) s respectively] (P < 0.05 or 0.01). (3) The percentage of effective search strategy increased significantly along with the test day in all groups except in the C57 lidocaine and apoE ischemia groups (P < 0.05 or 0.01), significantly lower in the ischemia and lidocaine groups than in the corresponding control groups (P < 0.05 or 0.01), significantly lower in the C57 lidocaine group than in the C57 ischemia group on the 4 th and 5 th days of test [25 (0-50)% vs. 50 (25-75)% and 37.5 (0-75)% vs. 50 (50-100)%], and in the apoE ischemia group than in the C57 ischemia group from the 3 rd to the 5th days of test [25 (0-25)% vs. 50 (25-75)%, 25 (0-25)% vs. 50 (25-75)%, and 25 (0-50)% vs. 50 (50-100)% respectively] (P < 0.05 or 0.01), and, however, significantly higher in the apoE lidocaine group than in the C57 lidocaine group [50 (0-75)% vs. 25 (0-50)% and 50 (25-100)% vs. 37.5 (0-75)%], and in the apoE lidocaine group than in apoE ischemia group [50 (0-75)% vs. 25 (0-25)% and 50 (25-100)% vs. 25 (0-50)%] on the 4 th and 5 th days of test (all P < 0.05). CONCLUSION: Transient global cerebral ischemia causes significant brain damage, which is more severe in the apoE mice than in the C57 mice. Lidocaine significantly worsens the ischemic brain damage in the C57 mice, and, however, significantly alleviates the ischemic cerebral result in the apoE mice.