ABSTRACT
Suanzaoren Tang， as documented in the Synopsis of the Golden Chamber （《金匮要略》），consists of Ziziphi Spinosae Semen， Poria， Anemarrhenae Rhizoma， Chuanxiong Rhizoma， and Glycyrrhizae Radix et Rhizoma， and is indicated for dysphoria， consumptive disease， and insomnia. In modern clinical practice，in addition to sleep disorders，Suanzaoren Tang and its modified formulas can also be used to treat anxiety disorders， as well as insomnia，cardiac intervention，myocardial infarction， and other diseases combined with anxiety. It is also used in combination with other Chinese medicinal formulas （such as Zhizichi Tang，Ganmai Dazaotang，Baihe Zhimu Dihuangtang，and Jinlingzi San），western medicines （such as paroxetine，zopiclone，enalapril，amlodipine，metformin，and sertraline hydrochloride）， acupuncture， and acupoint application to treat anxiety， as well as cardiovascular neurosis，cancer，hypertension，type 2 diabetes， and other diseases combined with anxiety. As revealed by clinical treatment results， Suanzaoren Tang and its modified formulas can significantly reduce anxiety scores such as Hamilton Anxiety Rating Scale（HAMA） and Self-rating Anxiety Scale（SAS），and improve anxiety symptoms with significant efficacy and few side effects. As reported by experimental pharmacological studies， Suanzaoren Tang can regulate the content of neurotransmitters such as dopamine（DA），homovanillic acid（HVA），γ-aminobutyric acid（GABA），noradrenaline （NE），5-hydroxyindoleacetic acid （5-HIAA），glutamic acid（Glu），β-endorphin（β-EP），5-hydroxytryptamine（5-HT）， and nitric oxide （NO）in the brain， and mediate immune function by reducing the levels of inflammatory cytokines such as interleukin-1β（IL-1β）and tumor necrosis factor-α（TNF-α），enhancing the proliferation of B lymphocytes，phagocytosis of peritoneal macrophages， and down-regulating the proliferation of T lymphocytes. Besides， it can also regulate the endocrine level to allow the homeostasis of nervous system， endocrine system， and immune system by increasing adrenocorticotropic hormone（ACTH）and corticosterone（CORT）levels and up-regulating glucocorticoid receptor（GR）expression， finally achieving the effect of anxiety prevention and treatment. The present study systematically reviewed the clinical and basic research progress on the prevention and treatment of anxiety with Suanzaoren Tang to provide references for the research on the mechanism and material basis of Suanzaoren Tang and the development of new drugs for anxiety.
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ObjectiveTo observe the effects of Suanzaoren Tang on the behavior， growth-associated protein-43 （GAP-43）， postsynaptic density protein-95 （PSD-95）， and synaptophysin Ⅰ （SynⅠ） of insomniac rats induced by p-chlorophenylalanine （PCPA）， and to investigate the mechanism of Suanzaoren Tang in improving the behavior of the insomniac rats. MethodSeventy-two SD rats were randomly assigned into 6 groups （12 rats in each group）： control group （normal saline）， PCPA （0.35 g·kg-1·d-1） group， estazolam （2.7×10-4 g·kg-1·d-1） group， and low-， medium-， and high-dose （3.25， 7.5， 15 g·kg-1·d-1， respectively） Suanzaoren Tang groups. The rat model of insomnia was established by intraperitoneal injection of PCPA and then the rats were administrated with corresponding drugs for 7 continuous days. The Morris water maze and Y-maze were used to test the learning and memory functions， and the open field to test anxiety. Histopathological changes in the hippocampus were observed via hematoxylin-eosin （HE） staining. The mRNA and protein levels of GAP-43， PSD-95， and SynⅠ in hippocampus were determined by real-time fluorescence quantitative polymerase chain reaction (Real-time PCR)， Western blot， and immunohistochemistry （IHC）. ResultCompared with the control group， the PCPA group showcased long escape latency， shortened time in the quadrants， and decreased times of crossing the platform in Morris water maze， decreased alternation correct rate was significantly Y-maze， and increased distance， mean velocity， and time in center of the open field test （P<0.01）. Furthermore， the PCPA-treated rats showed obvious pathological damage in the hippocampus and down-regulated mRNA and protein levels of GAP-43， PSD-95， and SynⅠ in hippocampus （P<0.01）. Compared with the PCPA group， the treatments with estazolam and different doses of Suanzaoren Tang improved the rat performance in Morris water maze， Y-maze， and open field test （P<0.05， P<0.01）， alleviated the hippocampal damage， and up-regulated the mRNA and protein levels of GAP-43， PSD-95， and SynⅠ （P<0.05， P<0.01）. ConclusionSuanzaoren Tang may alleviate the learning and memory disorders and anxiety in PCPA-induced insomnia rat model by up-regulating the mRNA and protein levels of hippocampal synaptic plasticity-associated proteins GAP-43， PSD-95， and SynⅠ.
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ObjectiveTo explore the effect of Suanzaoren Tang combined with Ziwu Liuzhu acupuncture on the vertebral artery hemodynamics， inflammatory cytokines， and neurotrophic factors in the patients with cervical insomnia with syndrome of deficiency of both heart and spleen. MethodThe random number table method was employed to assign 164 patients with cervical insomnia with syndrome of deficiency of both heart and spleen treated in the First Clinical Medical School of Guangzhou University of Chinese Medicine from January 2018 to June 2021 into a control group and an observation group. The control group was orally administrated with 1-2 mg estazolam tablets before bed for 4 weeks， and the observation group with Suanzaoren Tang combined with Ziwu Liuzhu acupuncture for 4 weeks. The therapeutic efficacy and safety were observed. The Pittsburgh Sleep Quality Index （PSQI） score， polysomnography monitoring results， hemodynamics parameters of vertebral artery， and serum levels of inflammatory cytokines and neurotrophic factors were compared before and after treatment. ResultExcept 4 dropouts， the remaining 160 patients were included in this study， with 80 patients in each group. The observation group had higher total effective rate than the control group ［92.50% （74/80） vs. 80.00% （64/80）， χ2=5.270， P<0.05］. Compared with that before treatment， the therapies in both groups decreased the PSQI score， sleep latency time， awakening time， awakening times， serum levels of interleukin-1β （IL-1β）， C-reactive protein （CRP）， and tumor necrosis factor-α （TNF-α） （P<0.01）. Meanwhile， they increased the proportion of rapid-eye-movement （REM） sleep， the diastolic blood flow velocity （Vd）， systolic blood flow velocity （Vs）， and mean blood flow velocity （MFV） of vertebral artery， as well as the serum levels of brain-derived neurotrophic factor （BDNF） and glial cell-derived neurotrophic factor （GDNF） （P<0.05， P<0.01）. Moreover， the observation group had lower PSQI score， sleep latency time， awakening time， awakening times， and serum IL-1β， CRP， and TNF-α levels （P<0.01） and higher proportion of REM sleep， Vd， Vs， MFV of vertebral artery， and serum BDNF and GDNF levels （P<0.05， P<0.01） than the control group. ConclusionZiwu Liuzhu acupuncture combined with Suanzaoren Tang can improve blood circulation of vertebral artery， reduce the serum levels of inflammatory cytokine， and increase the serum levels of neurotrophic factors to improve the sleep quality of the patients with cervical insomnia with syndrome of deficiency of both heart and spleen.
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ObjectiveTo explore the effect of Suanzaoren Tang combined with Ziwu Liuzhu acupuncture on the vertebral artery hemodynamics， inflammatory cytokines， and neurotrophic factors in the patients with cervical insomnia with syndrome of deficiency of both heart and spleen. MethodThe random number table method was employed to assign 164 patients with cervical insomnia with syndrome of deficiency of both heart and spleen treated in the First Clinical Medical School of Guangzhou University of Chinese Medicine from January 2018 to June 2021 into a control group and an observation group. The control group was orally administrated with 1-2 mg estazolam tablets before bed for 4 weeks， and the observation group with Suanzaoren Tang combined with Ziwu Liuzhu acupuncture for 4 weeks. The therapeutic efficacy and safety were observed. The Pittsburgh Sleep Quality Index （PSQI） score， polysomnography monitoring results， hemodynamics parameters of vertebral artery， and serum levels of inflammatory cytokines and neurotrophic factors were compared before and after treatment. ResultExcept 4 dropouts， the remaining 160 patients were included in this study， with 80 patients in each group. The observation group had higher total effective rate than the control group ［92.50% （74/80） vs. 80.00% （64/80）， χ2=5.270， P<0.05］. Compared with that before treatment， the therapies in both groups decreased the PSQI score， sleep latency time， awakening time， awakening times， serum levels of interleukin-1β （IL-1β）， C-reactive protein （CRP）， and tumor necrosis factor-α （TNF-α） （P<0.01）. Meanwhile， they increased the proportion of rapid-eye-movement （REM） sleep， the diastolic blood flow velocity （Vd）， systolic blood flow velocity （Vs）， and mean blood flow velocity （MFV） of vertebral artery， as well as the serum levels of brain-derived neurotrophic factor （BDNF） and glial cell-derived neurotrophic factor （GDNF） （P<0.05， P<0.01）. Moreover， the observation group had lower PSQI score， sleep latency time， awakening time， awakening times， and serum IL-1β， CRP， and TNF-α levels （P<0.01） and higher proportion of REM sleep， Vd， Vs， MFV of vertebral artery， and serum BDNF and GDNF levels （P<0.05， P<0.01） than the control group. ConclusionZiwu Liuzhu acupuncture combined with Suanzaoren Tang can improve blood circulation of vertebral artery， reduce the serum levels of inflammatory cytokine， and increase the serum levels of neurotrophic factors to improve the sleep quality of the patients with cervical insomnia with syndrome of deficiency of both heart and spleen.
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Objective:A comprehensive and in-depth analysis method for identification of chemical constituents in Suanzaoren Tang granules was established. Method:Ultra-performance liquid chromatography-quadrupole-time of flight mass spectrometry （UPLC-Q-TOF-MS/MS） was employed with the mobile phase of 0.1% formic acid aqueous solution （A）-acetonitrile （B） for gradient elution （0-8 min， 5%-17%B； 8-10 min， 17%B； 10-11 min， 17%-18%B； 11-12 min， 18%-20%B； 12-17 min， 20%-23%B； 17-22 min， 23%-33%B； 22-30 min， 33%-60%B； 30-32 min， 60%-100%B； 32-36 min， 100%B）， the flow rate of 0.3 mL·min<sup>-1</sup> and electrospray ionization （ESI）. High quality MS/MS data were scanned in positive and negative ion modes with scanning range of <italic>m</italic>/<italic>z</italic> 50-1 500. The local database of the chemical components from different Chinese medicines in Suanzaoren Tang granules was established by SCIEX OS software. Then the chemical components in Suanzaoren Tang granules were characterized by matching with the local database and comparing with the reference substance and literature information. Result:A total of 134 compounds were characterized and identified under positive and negative ion modes， mainly including flavonoids， triterpenoids， phthalides， steroidal saponins， alkaloids and organic phenolic acids. In addition， the sources of Chinese medicines for all compounds identified in Suanzaoren Tang granules were assigned. Among them， 41 were from Ziziphi Spinosae Semen， 11 were from Poria， 22 were from Anemarrhenae Rhizoma， 28 were from Chuanxiong Rhizoma and 35 were from Glycyrrhizae Radix et Rhizoma. Conclusion:The method can be used to identify the chemical constituents in Suanzaoren Tang granules systematically， quickly and accurately， which can provide a new strategy for the rapid and accurate identification of other Chinese patent medicines.
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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<sup>-4</sup> g·kg<sup>-1</sup>·d<sup>-1</sup>）， low-dose Suanzaoren Tang group（4.59 g·kg<sup>-1</sup>·d<sup>-1</sup>）and high-dose Suanzaoren Tang group （18.36 g·kg<sup>-1</sup>·d<sup>-1</sup>）. 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.
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Sleep has been widely concerned by the medical field all over the world. Sleep deprivation can cause damage to organs of the human body， which is related to the occurrence of a variety of diseases. Besides， the pathological change in different organs of the human body is also a key factor that causes or aggravates insomnia. When treating insomnia and its complications， traditional Chinese medicine （TCM） focuses on the homology of the brain and heart， and insomnia is mainly treated from the five internal organs， especially the heart and liver. Sleep duration and structure change with age. The prevalence of insomnia is higher in older individuals susceptible to complications than in the younger population. In TCM， insomnia of blood deficiency and Yin deficiency is common among the elderly. Suanzaoren Tang is a classic prescription for nourishing blood and calming the mind and it is critical in the treatment of "sleeplessness due to consumptive disease and dysphoria"， with the effects of nourishing liver blood to calm the mind and clearing internal heat to relieve dysphoria. It has good efficacy on the insomnia of the elderly caused by deficiency of Qi and blood and abnormal operation of nutrient Qi and defense Qi. Furthermore， it also shows a certain therapeutic effect on insomnia combined with cardiovascular and cerebrovascular diseases. The present study revealed the damage to the brain， heart， and liver caused by sleep deprivation and the effect of Suanzaoren Tang on the brain， heart， and liver， and clarified the facts that Suanzaoren Tang inhibited the damage to organs caused by sleep deprivation and regulated energy metabolism， thereby exploring the sedative and hypnotic mechanism of Suanzaoren Tang to provide new ideas for Suanzaoren Tang in the treatment of sleep disorders and other diseases.
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Objective:To study the effect of Suanzaoren Tang on energy metabolism of liver mitochondria in aged rats with chronic rapid eye movement （REM） sleep deprivation. Method:Fifty male Wistar rats were randomly divided into the control group， model group， estazolam group （0.18 mg·kg<sup>-1</sup>·d<sup>-1</sup>）， and low- （6.48 g·kg<sup>-1</sup>·d<sup>-1</sup>） and high-dose （12.96 g·kg<sup>-1</sup>·d<sup>-1</sup>） Suanzaoren Tang groups. Rats in all groups except for the control group received subcutaneous injection of <italic>D</italic>-galactose and then were deprived of sleep using the multiple platform method after the last administration. Following successful modeling， the rats in each group were treated with intragastric administration of the corresponding drugs for seven consecutive days. The morphology of liver mitochondria was observed under the transmission electron microscope. The content of adenosine triphosphate （ATP） in rat liver was detected by colorimetry， and the activities of ubiquinone oxidoreductase （complex Ⅰ）， succinate-ubiquinone oxidoreductase （complex Ⅱ）， ubiquinol-cytochrome c oxidoreductase （complex Ⅲ）， and cytochrome c oxidase （complex Ⅳ） in mitochondrial respiratory chain of rat liver were measured by colorimetry. The protein expression levels of citrate synthase （CS）， isocitrate dehydrogenase （IDH）， and ATP synthase， H<sup>+</sup> transporting， mitochondrial F0 complex， subunit b， isoform 1 （ATP5F1） in rat liver were assayed by Western blot. Result:The mitochondrial damage in rat liver of the model group was more serious than that in the control group， manifested as mitochondrial swelling and deformation as well as cristae rupture and reduction. The comparison with the model group revealed that both the positive control and Suanzaoren Tang at the high dose obviously alleviated the mitochondrial swelling and deformation and reduced cristae rupture， with better improvements observed in the high-dose Suanzaoren Tang group. Compared with the control group， the content of ATP， the activities of mitochondrial respiratory chain complexes Ⅰ， Ⅱ， Ⅲ， and Ⅳ， and the protein expression levels of IDH， CS， and ATP5F1 in rat liver of the model group were all significantly decreased （<italic>P<</italic>0.01）. Compared with the model group， the content of ATP， the activities of mitochondrial respiratory chain complexes Ⅰ， Ⅱ， Ⅲ， and Ⅳ， and the protein expression levels of IDH， CS， and ATP5F1 in rat liver of the high-dose Suanzaoren Tang group were all significantly increased （<italic>P<</italic>0.05，<italic>P<</italic>0.01）. In the positive control group， the content of ATP， the activities of mitochondrial respiratory chain complexes I and Ⅲ， and the protein expression levels of CS and ATP5F1 in rat liver were significantly increased （<italic>P<</italic>0.05，<italic> P<</italic>0.01）. The activities of mitochondrial respiratory chain complexes Ⅰ and Ⅲ and the ATP5F1 protein expression in the low-dose Suanzaoren Tang group were significantly elevated （<italic>P<</italic>0.05， <italic>P<</italic>0.01）. Conclusion:Suanzaoren Tang alleviates the abnormal liver energy metabolism induced by chronic REM sleep deprivation in the elderly rats， which may be related to its enhancement of mitochondrial electron transport chain enzyme activities and the up-regulation of protein expression levels of CS， IDH and ATP5F1.
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Objective:To investigate the effect of Suanzaoren Tang on mitochondria-mediated neuronal apoptosis. Method:Fifty male Wistar rats were randomly divided into the control group， model group， estazolam group （0.18 mg·kg<sup>-1</sup>·d<sup>-1</sup>）， and low- （6.48 g·kg<sup>-1</sup>·d<sup>-1</sup>） and high-dose （12.96 g·kg<sup>-1</sup>·d<sup>-1</sup>） Suanzaoren Tang groups. Rats in all groups except for the control group received subcutaneous injection of <italic>D</italic>-galactose and then were deprived of sleep using the multiple platform method after the last administration. Following successful modeling， the rats in each group were treated with intragastric administration of the corresponding drugs for seven consecutive days. The morphology of mitochondria in hypothalamus was observed under a transmission electron microscope. The activities of Na<sup>+</sup>-K<sup>+</sup>-ATPase and Ca<sup>2+</sup>-Mg<sup>2+</sup>-ATPase in hypothalamus were detected by spectrophotometry. Western blotting was conducted to determine the protein expression levels of cytochrome C （Cyt C）， B cell lymphoma-2 （Bcl-2）， Bcl-2 associated X protein （Bax） and cysteinyl aspartate-specific protease-3 （Caspase-3） in hypothalamus. Result:In the control group， there was no obvious pathological change in mitochondria， which were moderate in size and oval or spindle in shape， with the cristae arranged orderly. Compared with the control group， the model group exhibited abnormal mitochondrial morphology， manifested as obvious swelling， vacuolation， myelin figures， and cristae rupture and reduction. The comparison with the model group revealed that both the estazolam group and high-dose Suanzaoren Tang group alleviated the mitochondrial damage and reduced the vacuolation and swelling. Only some cristae rupture was present. The improvements were more obvious in the high-dose Suanzaoren Tang group. Compared with the control group， the activities of Na<sup>+</sup>-K<sup>+</sup>-ATPase and Ca<sup>2+</sup>-Mg<sup>2+</sup>-ATPase and the Bcl-2 protein expression in the model group were significantly decreased （<italic>P<</italic>0.01）， whereas the protein expression levels of Cyt C， Bax， and Caspase-3 were significantly increased （<italic>P</italic><0.01）. Compared with the model group， the activities of Na<sup>+</sup>-K<sup>+</sup>-ATPase and Ca<sup>2+</sup>-Mg<sup>2+</sup>-ATPase and the Bcl-2 protein expression in the estazolam group and the high-dose Suanzaoren Tang group were significantly elevated （<italic>P<</italic>0.01）， while the protein expression levels of Cyt C， Bax， and Caspase-3 were significantly lowered （<italic>P<</italic>0.05， <italic>P<</italic>0.01）. The activity of Na<sup>+</sup>-K<sup>+</sup>-ATPase and the Bcl-2 protein expression in the low-dose Suanzaoren Tang group were increased significantly （<italic>P<</italic>0.01）， but the Bax protein expression was down-regulated （<italic>P<</italic>0.05）. Conclusion:Suanzaoren Tang is able to improve the mitochondrial function of hypothalamic nerve cells and inhibit their apoptosis.
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Objective:To study the mechanism of Suanzaoren Tang in regulating the energy metabolism of myocardial mitochondria in aged rats with chronic rapid eye movement （REM） sleep deprivation through the sirtuin 3 （SIRT3）/superoxide dismutase2 （SOD2） signaling pathway. Method:Fifty male Wistar rats were randomly divided into the control group， model group， estazolam group （0.18 mg·kg<sup>-1</sup>·d<sup>-1</sup>）， and low- （6.48 g·kg<sup>-1</sup>·d<sup>-1</sup>） and high-dose （12.96 g·kg<sup>-1</sup>·d<sup>-1</sup>） Suanzaoren Tang groups. Rats in all groups except for the control group received subcutaneous injection of <italic>D</italic>-galactose and then were deprived of sleep using the multiple platform method after the last administration. Following successful modeling， the rats in each group were treated with intragastric administration of the corresponding drugs for seven consecutive days. The morphology of myocardial mitochondria was observed under a transmission electron microscope. The content of adenosine triphosphate （ATP） in rat hypothalamus was detected by colorimetry， while the malondialdehyde （MDA） content and the SOD activity in myocardium were measured by spectrophotometry. Real-time polymerase chain reaction （Real-time PCR） and Western blot were conducted to determine the mRNA and protein expression levels of SIRT3 and SOD2 in rat myocardium. The localization of SIRT3 was detected by immunofluorescence staining. Result:Compared with the control group， the model group exhibited a disordered arrangement of myocardial filaments， accompanied by filament rupture and dissolution， obviously swollen mitochondria arranged in disorder， and blurring and even rupture of most mitochondrial cristae. Besides， the content of ATP and SOD activity in the myocardium decreased significantly （<italic>P<</italic>0.01）， whereas that of MDA increased significantly （<italic>P<</italic>0.01）. The mRNA and protein expression levels of SIRT3 and SOD2 were down-regulated significantly （<italic>P<</italic>0.01）， and the average fluorescence intensity of SIRT3 protein declined significantly （<italic>P<</italic>0.01）. The comparison with the model group revealed that high-dose Suanzaoren Tang enabled the myocardial filaments to be neatly arranged， relieved the mitochondrial damage and swelling， only manifested as partial mitochondrial cristae rupture， significantly increased ATP content， SOD activity， as well as SIRT3 and SOD2 mRNA and protein expression levels （<italic>P<</italic>0.01）， reduced the content of MDA （<italic>P<</italic>0.01）， and enhanced the average fluorescence intensity of SIRT3 protein （<italic>P<</italic>0.05）. The myocardial mitochondrial injury in the estazolam group was also alleviated. The activity of SOD and the SIRT3 and SOD2 mRNA and protein expression levels in the myocardium were significantly elevated （<italic>P<</italic>0.01）， while the activity of MDA was significantly lowered （<italic>P<</italic>0.01）. In the low-dose Suanzaoren Tang group， the improvement in myocardial mitochondrial injury was not obvious. However， both the SOD activity and SOD2 protein expression were significantly increased （<italic>P<</italic>0.05）. Conclusion:Suanzaoren Tang ameliorates the myocardial mitochondria injury and abnormal energy metabolism induced by chronic REM sleep deprivation in aged rats possibly by up-regulating the SIRT3 and SOD2 expression.
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Objective:To observe the clinical therapeutic effect of Suanzaoren Tang combined with fluoxetine in the treatment of patients with depression of liver stagnation and blood deficiency accompanied by insomnia. Method:The patients with depression of liver stagnation and blood deficiency accompanied by insomnia （120 cases） were randomly divided into an observation group and a control group， with 60 cases in each group. The patients in the observation group received Suanzaoren Tang combined with fluoxetine， and those in the control group received fluoxetine. The course of treatment was eight weeks. The clinical efficacy was evaluated with Hamilton Depression Rating Scale （HAMD）， Pittsburgh Sleep Quality Index（PSQI）， and Activities of Daily Living （ADL） score. Enzyme-linked immunosorbent assay （ELISA） was used to determine the plasma levels of 5-hydroxytryptamine （5-HT）， norepinephrine （NE），brain-derived neurotrophic factor （BDNF）， glial cell-derived neurotrophic factor （GDNF）， neuron-specific enolase （NSE）， and S100<italic>β</italic>. Result:After eight weeks of treatment， the scores of HAMD and PSQI were reduced（<italic>P</italic><0.01）， while the scores of ADL were elevated（<italic>P</italic><0.01），and the levels of 5-HT， NE， GDNF and BDNF were up-regulated （<italic>P</italic><0.01） in the plasma of patients in the observation group as compared with those before treatment. After treatment， compared with the control group， the observation group showed increased total effective rate（<italic>P</italic><0.01）， decreased scores of HAMD and PSQI （<italic>P</italic><0.01）， elevated score of ADL（<italic>P</italic><0.01）， up-regulated levels of 5-HT， NE， GDNF and BDNF in plasma， and declining NSE and S100<italic>β</italic>（<italic>P</italic><0.01）. Conclusion:Suanzaoren Tang combined with fluoxetine is superior to fluoxetine alone in treating the depression of liver stagnation and blood deficiency accompanied by insomnia. Its therapeutic effect is achieved by increasing the release of monoamine neurotransmitters and promoting the secretion of BDNF and GDNF in the brain.
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Objective::To explore the mechanism of Suanzaoren Tang in improving learning and memory impairment induced by sleep deprivation based on Toll-like receptor (TLR4)/nuclear transcription factor-κB (NF-κB) signaling pathway. Method::The experimental rats were randomly divided into blank group, model group, melatonin group (2.5×10-4 g·kg-1·d-1) and Suanzaoren Tang group (12.96 g·kg-1·d-1). Except the blank group, the chronic sleep deprivation model was established in other groups. After 28 days of continuous administration, the learning and memory of the rats were assessed by Morris water maze. The expressions of interleukin-1β (IL-1β), interleukin-6 (IL-6) and tumor necrosis factor-α(TNF-α)in serum were detected by enzyme-linked immunosorbent assay (ELISA). The mRNA expression levels of TLR4, NF-κB p65 and nuclear transcription factor inhibitory protein α (IκBα) in the hippocampus of rats were detected by real-time fluorescence quantitative polymerase chain reaction (Real-time PCR), and the protein expressions of TLR4, IκBα, p-IκBα and NF-κB p65 in the hippocampus of rats were detected by immunohistochemistry (IHC). Result::Compared with the blank group, the platform latency, total swimming distance and the first landing time of the model group increased significantly (P<0.01), while the number of crossing platforms and target quadrant time decreased significantly (P<0.01). Compared with the model group, the platform latency, total swimming distance and the first platform time were reduced in the melatonin group and the Suanzaoren group (P<0.05, P<0.01), while the number of crossing platforms and target quadrants time were increased(P<0.05, P<0.01). Compared with the blank group, the expression levels of TNF-α, IL-1β and IL-6 in the model group were significantly increased (P<0.01). Compared with the model group, the levels of TNF-α, IL-1β and IL-6 in the serum of melatonin group and Suanzaoren Tang group decreased (P<0.05, P<0.01). Compared with the blank group, mRNA and protein expression levels of TLR4 and NF-κB p65 in the hippocampus of the model group were increased (P<0.01), while mRNA and protein expression levels of IκBα were decreased (P<0.01). Compared with the model group, the mRNA and protein expressions of TLR4 and NF-κB p65 in the hippocampus of melatonin group and Suanzaoren Tang group decreased (P<0.05, P<0.01), while the mRNA and protein expression levels of IκBα increased (P<0.05, P<0.01). Compared with the blank group, the protein expression level of p-IκBα in the hippocampus of the model group was significantly increased (P<0.01). Compared with the model group, the protein expression level of IκBα in the hippocampus of melatonin group and Suanzaoren Tang group was increased (P<0.05, P<0.01). Conclusion::Suanzaoren Tang can improve learning and memory impairment induced by sleep deprivation in rats, and its mechanism may be related to its inhibition of TLR4/NF-κB signaling pathway in hippocampus.
ABSTRACT
Objective:To study the effect of modified Xiao Chaihutang on the expressions of excitatory amino acid transporters（EAATs） and vesicle glutamate transporters（VGLUTs）in hippocampus of rats with chronic depression， in order to explore the anti-depressant mechanism of modified Xiao Chaihutang based on glutamate transport. Method:A total of 120 SD rats were randomly divided into normal group， model group， and low， middle and high-dose modified Xiaochaihutang groups （6.5， 13， 26 g·kg-1） and riluzole group， with 20 rats in each group．Except normal group， the depression model of rats was prepared through Chronic restraint stress（CRS）. The normal group and the model group were intragastrically （ig） given normal saline. The modified Xiao Chaihutang groups were intragastrically given corresponding herbal drugs （6.5， 13， 26 g·kg-1）， and the Riluzole group was given Riluzole 20 mg·kg-1 through intraoeritoneal injection for 21 days， once a day. Then the depressive behaviors of rats were observed by forced swimming test （FST） and tail suspension test （TST）. The level of glutamic acid （Glu） in rats hippocampus was determined by high performance liquid chromatography （HPLC）. The mRNA expressions of EAAT1， EAAT2 and EAAT3 in hippocampus were detected by Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）method. Western blot was used to detect the protein expressions of EAAT1， EAAT2， EAAT3， VGLUT1 and VGLUT2 in rat hippocampus tissue. Nissl staining was used to observe the morphology of hippocampal neurons in rats. Immunohistochemical（IHC）S-P method were used to detect the location expressions of EAAT1， EAAT2 and NeuN proteins in rat hippocampal CA1 region tissue. Result:The immobility times in FST and TST were increased significantly（P<0.01）， the mRNA and protein expressions of EAAT1，EAAT2，EAAT3 were decreased significantly （P<0.01）， and as well as the expressions of VGLUT1 and NeuN were decreased significantly（P<0.01）， while the level of Glutamate and the expression of VGLUT2 were increased significantly（P<0.01） in model group， compared with normal group. Compared with model group，the immobility times in FST and TST were decreased significantly（P<0.05， P<0.01）， mRNA and protein expressions of EAAT1，EAAT2，EAAT3 were increased significantly（P<0.01）， and expressions of VGLUT1 and NeuN were increased significantly（P<0.01）. However， the level of Glutamate and the expression of VGLUT2 were decreased significantly（P<0.01）， and the damage of hippocampal neurons in rats was mild in middle and high-dose modified Xiao Chaihutang groups. Conclusion:Modified Xiao Chaihutang has an anti-depressive effect. Its mechanism may be related to its up-regulation of expressions of EAAT1， EAAT2， EAAT3 genes and VGLUT1 protein in the hippocampus of depression model rats.
ABSTRACT
Objective:To study the effect of modified Suanzaoren Tang on the expression of excitatory amino acids receptor（EAARs） in hippocampus of rats with chronic depression， and to explore the anti-depressant mechanism of modified Suanzaoren Tang based on excitatory amino acids receptor. Method:Sixty SD rats were randomly divided into normal group，model group，and low，middle and high-dose modified Suanzaoren Tang groups，and ketamine group，with 10 rats in each group．Except normal group，the depression model of rats was prepared by using chronic restraint stress（CRS）.The normal group and model group were intragastrically（ig） given normal saline．the modified Suanzaoren Tang groups were intragastrically given corresponding herbal drugs 6，12，24 g·kg-1， ketamine group group were given ketamine 0.015 g·kg-1 through intraoeritoneal injection，for 21 days，once a day．Then the depressive behaviors of rats were observed by Morris water maze and novelty feeding experiment.Western blot was used to detect the levels of DAR1，NMDAR2A，NMDAR2B，GluR1，mGluR1，CaMKⅡα and CaMKⅡβ protein expression in rat hippocampus tissue. Result:Compared with normal group，the time of novel ingestion and escape latencywere prolonged significantly（P<0.01）， and the time of space exploration was shortened significantly（P<0.01）.The levels of NMDAR1，NMDAR2A，NMDAR2B，mGluR1 and CaMKⅡβ expression were increased significantly（P<0.01），while the levels of GluR1 and CaMKⅡα expression were decreased significantly（P<0.01）in model group. Compared with model group，the time of novel ingestion and escape latency were shortened significantly （P<0.01）， and the time of space exploration was prolonged significantly（P<0.01）.The levels of NMDAR1，NMDAR2A，NMDAR2B，mGluR1 and CaMKⅡβ protein expression were decreased significantly（P<0.01），but the levels of GluR1 and CaMKⅡα expression were increased decreased significantly（P<0.01）in middle and high-dose modified Suanzaoren Tang groups. Conclusion:Modified Suanzaoren Tang can improve the behavior of chronic depression rats effectly. Its mechanism may be related with reduction the expression of NMDAR1，NMDAR2A，NMDAR2B，mGluR1 and CaMKⅡβ protein ，increase the expression of GluR1and CaMKⅡα protein.
ABSTRACT
Objective:To observe the effect of Suanzaoren Tang on hippocampal neuroinflammation in APP/PS1 mice and to explore its possible mechanism of neuroprotection. Method:The mice were randomly divided into blank group, model group, donepezil group(0.92 mg·kg-1), Suanzaoren Tang low and high-dose groups(12.96,25.92 g·kg-1). After 30 days of continuous administration in each group, pathological changes of dentate gyrus (DG) in hippocampus of mice in each group were observed by Nissl staining.The levels of tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) in serum of each group were detected by enzyme-linked immunosorbent assay (ELISA). Real-time quantitative polymerase chain reaction (Real-time PCR) was used to detect the mRNA expression levels of TNF-α and IL-1β in hippocampus of each group. The expression levels of glial fibrillary acidic protein (GFAP) and ionic calcium binding protein 1 (IBA1) in hippocampus of each group were detected by immunohistochemical staining (ICH) and Western blot. Result:Compared with blank group, the granule cells in DG region were unevenly arranged in model group, with obvious cell loss, and the nissl bodies in some neurons disappeared or condensed, serum TNF-α and IL-1β content significantly increased (Pα and IL-1β mRNA expression quantity significantly increased (PPα and IL-1β in the serum were decreased(PPα and IL-1β mRNA in the hippocampus were decreased(PPPPConclusion:Suanzaoren Tang can improve neuronal loss in APP/PS1 double transgenic mice, and its mechanism may be related to the regulation of hippocampal neuroinflammation in mice.