Objectives This study aimed to investigate the therapeutic effects of Suan Zao Ren(Semen Ziziphi Spinosae,SZS)extract on insomnia induced by p-chlorophenylalanine(PCPA)in rats and its influence on the thioredoxin-inte...Objectives This study aimed to investigate the therapeutic effects of Suan Zao Ren(Semen Ziziphi Spinosae,SZS)extract on insomnia induced by p-chlorophenylalanine(PCPA)in rats and its influence on the thioredoxin-interacting protein(TXNIP)/nucleotide-binding domain Leucine-rich repeat and pyrin domain-containing receptor 3(NLRP3)inflammasome pathway,and to preliminarily explore themechanism by which SZS extract improves insomnia.Methods Fifty male Sprague–Dawley(SD)rats were used,with 8 rats in the blank group and 42 rats in the modeling group.The modeling group was induced by intraperitoneal injection of PCPA at a dose of 500mg·kg^(-1) for six consecutive days,with daily cage exchange.After 6 days,40 successfully modeled rats were randomly divided into five groups:themodel group(equal volume of distilled water),the positive group(0.75 mg·kg^(-1)),and low-,medium-,and high-dose SZS extract groups(1.5,3,and 6 g·kg^(-1),respectively),with 8 rats in each group.Treatments were administered for seven consecutive days.Enzyme-linked immunosorbent assay was used to measure levels of 5-hydroxytryptamine(5-HT)and gamma-aminobutyric acid(GABA)in the rat cerebral cortex.The thiobarbituric acid(TBA)method was used to determine malondialdehyde(MDA)levels,and the hydroxylamine method was used to determine superoxide dismutase(SOD)levels.The 2,2′-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid)method was used to measure total antioxidant capacity(TAOC)in the cerebral cortex.Pathological changes in the cerebral cortex were observed,and Western blot was used to detect the protein expressions of TXNIP,NLRP3,apoptosis-associated speck-like protein containing a Caspase activation and recruitment domain(CARD),and cysteine–aspartate-specific protease 1(Caspase-1)in the cerebral cortex.Results Compared with the blank group,the model group showed a significantly prolonged sleep latency(p<0.001)and a significantly shortened sleep duration(p<0.001).There were no changes in serum MDA and SOD levels.MDA levels in the cerebral cortex were significantly increased(p<0.001),while SOD and TAOC levels were significantly decreased(p<0.001).The 5-HT level was increased(p<0.05),and the GABA level was significantly decreased(p<0.001).SZS extract improved these conditions to varying degrees.Light microscopy showed no significant changes in cortical neurons but transmission electron microscopy revealed intact mitochondrial structures in the blank group,while the model group showed swollen and unclear mitochondria with reduced organelles.After 7 days of treatment,these conditions improved in the SZS extract groups.Compared with the blank group,the expressions of the four proteins in the model group were increased,and the expressions of these proteins were decreased in the SZS extract groups compared with the model group.Conclusion SZS extract may exert an antioxidant effect to treat insomnia by downregulating the expression of TXNIP/NLRP3 proteins and regulating oxidative stress levels in the cerebral cortex.展开更多
Conventional quantity determination of Chinese herbal medicines by High-Performance Thin-Layer Chromatography (HPTLC) must be carried out on a TLC scanner for measuring the marker component on the HPTLC plate. General...Conventional quantity determination of Chinese herbal medicines by High-Performance Thin-Layer Chromatography (HPTLC) must be carried out on a TLC scanner for measuring the marker component on the HPTLC plate. Generally the HPTLC image of the herbal drug always takes the photograph as the permanent visual document. Considering the photo of the HPTLC image preserving all the visual quality-related parameters, there might be an alternative cost/effective approach worth studying. It would be presumably applicable to use the exclusively developed software as a virtual scanner instead of the TLC Scanner instrument to do digital scanning, integrating and calculating target components in the photo of the HPTLC image. It means digitally scanning the photo of the HPTLC image solely with software does the same thing as the instrumental scanner does. Based on this protocol, we primarily conducted the assay of the contents of the major saponins, jujuboside a and jujuboside b, in Ziziphi Spinosae Semen (Suan-Zao-Ren). Using home-developed software “digiscan” for acquiring the raw data and integrating the peaks area of the target components with “Origin Pro”, the contents of jujuboside a and jujuboside b can then be calculated respectively. The primary investigated experiment results were well compatible with official HPLC-ELSD method in Chinese Pharmacopoeia. The methodology validation and comparative analysis with HPLC confirmed that the unprecedented method is acceptable. The advantage of the recommended approach is far more simple, flexible and cost/effective than the conventional method, and it can be done at anytime and anywhere once only if the high quality HPTLC image photo is available. There would be a bright perspective for application to other herbal medicines. And the one-stop software for merging digital scanning, integration and calculation together is in progress.展开更多
基金Research on the Mechanism of Suan Zao Ren’s(SZS)Hypnotic Effect Based on TXNIP/NLRP3 Signaling Pathway(2023-ZXFZJJ-JW-071).
文摘Objectives This study aimed to investigate the therapeutic effects of Suan Zao Ren(Semen Ziziphi Spinosae,SZS)extract on insomnia induced by p-chlorophenylalanine(PCPA)in rats and its influence on the thioredoxin-interacting protein(TXNIP)/nucleotide-binding domain Leucine-rich repeat and pyrin domain-containing receptor 3(NLRP3)inflammasome pathway,and to preliminarily explore themechanism by which SZS extract improves insomnia.Methods Fifty male Sprague–Dawley(SD)rats were used,with 8 rats in the blank group and 42 rats in the modeling group.The modeling group was induced by intraperitoneal injection of PCPA at a dose of 500mg·kg^(-1) for six consecutive days,with daily cage exchange.After 6 days,40 successfully modeled rats were randomly divided into five groups:themodel group(equal volume of distilled water),the positive group(0.75 mg·kg^(-1)),and low-,medium-,and high-dose SZS extract groups(1.5,3,and 6 g·kg^(-1),respectively),with 8 rats in each group.Treatments were administered for seven consecutive days.Enzyme-linked immunosorbent assay was used to measure levels of 5-hydroxytryptamine(5-HT)and gamma-aminobutyric acid(GABA)in the rat cerebral cortex.The thiobarbituric acid(TBA)method was used to determine malondialdehyde(MDA)levels,and the hydroxylamine method was used to determine superoxide dismutase(SOD)levels.The 2,2′-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid)method was used to measure total antioxidant capacity(TAOC)in the cerebral cortex.Pathological changes in the cerebral cortex were observed,and Western blot was used to detect the protein expressions of TXNIP,NLRP3,apoptosis-associated speck-like protein containing a Caspase activation and recruitment domain(CARD),and cysteine–aspartate-specific protease 1(Caspase-1)in the cerebral cortex.Results Compared with the blank group,the model group showed a significantly prolonged sleep latency(p<0.001)and a significantly shortened sleep duration(p<0.001).There were no changes in serum MDA and SOD levels.MDA levels in the cerebral cortex were significantly increased(p<0.001),while SOD and TAOC levels were significantly decreased(p<0.001).The 5-HT level was increased(p<0.05),and the GABA level was significantly decreased(p<0.001).SZS extract improved these conditions to varying degrees.Light microscopy showed no significant changes in cortical neurons but transmission electron microscopy revealed intact mitochondrial structures in the blank group,while the model group showed swollen and unclear mitochondria with reduced organelles.After 7 days of treatment,these conditions improved in the SZS extract groups.Compared with the blank group,the expressions of the four proteins in the model group were increased,and the expressions of these proteins were decreased in the SZS extract groups compared with the model group.Conclusion SZS extract may exert an antioxidant effect to treat insomnia by downregulating the expression of TXNIP/NLRP3 proteins and regulating oxidative stress levels in the cerebral cortex.
文摘Conventional quantity determination of Chinese herbal medicines by High-Performance Thin-Layer Chromatography (HPTLC) must be carried out on a TLC scanner for measuring the marker component on the HPTLC plate. Generally the HPTLC image of the herbal drug always takes the photograph as the permanent visual document. Considering the photo of the HPTLC image preserving all the visual quality-related parameters, there might be an alternative cost/effective approach worth studying. It would be presumably applicable to use the exclusively developed software as a virtual scanner instead of the TLC Scanner instrument to do digital scanning, integrating and calculating target components in the photo of the HPTLC image. It means digitally scanning the photo of the HPTLC image solely with software does the same thing as the instrumental scanner does. Based on this protocol, we primarily conducted the assay of the contents of the major saponins, jujuboside a and jujuboside b, in Ziziphi Spinosae Semen (Suan-Zao-Ren). Using home-developed software “digiscan” for acquiring the raw data and integrating the peaks area of the target components with “Origin Pro”, the contents of jujuboside a and jujuboside b can then be calculated respectively. The primary investigated experiment results were well compatible with official HPLC-ELSD method in Chinese Pharmacopoeia. The methodology validation and comparative analysis with HPLC confirmed that the unprecedented method is acceptable. The advantage of the recommended approach is far more simple, flexible and cost/effective than the conventional method, and it can be done at anytime and anywhere once only if the high quality HPTLC image photo is available. There would be a bright perspective for application to other herbal medicines. And the one-stop software for merging digital scanning, integration and calculation together is in progress.
