1.Mechanism of Neochlorogenic Acid in Ameliorating Psoriatic Keratinocyte Proliferation and Inflammation by Targeting HSP90 to Modulate NF-κB/NLRP3 Signaling Pathway
Mengyao JIANG ; Xinwei ZHANG ; Bin YANG ; Ping SONG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(17):89-98
ObjectiveTo investigate the target proteins directly bound by neochlorogenic acid (NA) and the molecular mechanisms that ameliorate the proliferation and inflammatory response of psoriatic keratinocytes. MethodsM5-induced HaCaT cells were used as a psoriatic keratinocyte proliferation and inflammatory cell model. The synthesized NA probe (NA-P) and NA prodrug were first evaluated for cell viability using a cell proliferation/cell counting kit-8(CCK-8). The potency of NA and NA-P was evaluated in the safe concentration range, and the effects of 0-100 μmol·L-1 NA and probe on M5-induced proliferation of HaCaT cells were detected using CCK-8. The effects of 20, 40, 80 μmol·L-1 NA and 80 μmol·L-1 NA-P on the expression of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), interleukin-23 (IL-23), and interleukin-17A (IL-17A) inflammatory factors were detected by enzyme-linked immunosorbent assay (ELISA), and real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) was used to measure the effects of NA on the mRNA expression of keratin 16 (K16) in HaCaT cells, S100 calcium-binding protein A9 (S100A9), S100 calcium-binding protein A7 (S100A7), IL-6, IL-17A, and chemokine 1 (CXCL1). In vitro fluorescence labeling and competition experiments using NA-P were performed, and target protein angling and analysis using pull-down experiments combined with liquid chromatography-mass spectrometry (Pull-down/LC-MS/MS) were conducted. Target validation was performed using pull-down experiments combined with protein immunoblotting (Pull down-WB), cellular heat transfer analysis combined with protein immunoblot (CETSA-WB) experiments, and molecular docking. Finally, Real-time PCR was utilized to detect the effects of 20, 40, 80 μmol·L-1 NA and 80 μmol·L-1 NA-P on the mRNA expression of IL-1β, nucleotide-binding oligomeric structural domain-like receptor protein 3 (NLRP3), apoptosis-associated speckled-like protein (ASC), and cysteine protease-1 (Caspase-1) in HaCaT cells. Protein immunoblot (Western blot) was used to detect the effects of phosphorylated p65 (p-p65), p65, phosphorylated human nuclear factor-κB inhibitory protein α (p-IκBα), human nuclear factor κB inhibitory protein α (IκBα), and heat shock protein 90 (HSP90) expression. ResultsIn the 200 μmol·L-1 safe concentration range, HaCaT cell proliferation, increased expression of TNF-α, IL-1β, IL-23, and IL-17A inflammatory factors, and increased mRNA expression of K16, S100A9, S100A7, IL-6, IL-17A, and CXCL1 were observed in the M5 group compared with the blank group. Cell proliferation in 5-100 μmol·L-1 NA and NA-P groups was inhibited, and the expression of TNF-α, IL-1β, IL-23, and IL-17A inflammatory factors was decreased in the NA-L, NA-M, NA-H, and NA-P-H groups. The mRNA expression of K16, S100A9, S100A7, IL-6, IL-17A, and CXCL1 was decreased (P<0.05). High-confidence targets were screened for HSP90 protein by Pull-down/LC-MS/MS using 200 μmol·L-1 NA competing with 100 μmol·L-1 NA-P. Compared with that in the blank group, the mRNA expression of NLRP3, IL-1β, ASC, and Caspase-1, as well as the expression of p-p65/p65, p-IκBα/IκBα, and HSP90 protein, were increased in HaCaT cells in the M5 group (P<0.05). Compared with that in the M5 group, the mRNA expression of NLRP3, IL-1β, ASC, and Caspase-1 of cells in the NA-L group, the NA-M group, the NA-H group, and the NA-P-H group was decreased (P<0.05). p-p65/p65 and p-IκBα/IκBα were decreased in the NA-M and NA-H groups (P<0.05), and there was no change in HSP90 protein. Pull down-WB showed that NA could directly target HSP90 protein, and NA binding to HSP90 protein enhanced its thermal stability. Molecular docking of NA with HSP90 family proteins HSP90AA1, HSP90B1, and HSP90AB1 all resulted in highly stable binding. ConclusionNA can inhibit the proliferation and inflammatory response of psoriatic keratinocytes by a mechanism that may be achieved by targeting HSP90 to modulate the NF-κB/NLRP3 signaling pathway.
2.Mechanism of Neochlorogenic Acid in Ameliorating Psoriatic Keratinocyte Proliferation and Inflammation by Targeting HSP90 to Modulate NF-κB/NLRP3 Signaling Pathway
Mengyao JIANG ; Xinwei ZHANG ; Bin YANG ; Ping SONG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(17):89-98
ObjectiveTo investigate the target proteins directly bound by neochlorogenic acid (NA) and the molecular mechanisms that ameliorate the proliferation and inflammatory response of psoriatic keratinocytes. MethodsM5-induced HaCaT cells were used as a psoriatic keratinocyte proliferation and inflammatory cell model. The synthesized NA probe (NA-P) and NA prodrug were first evaluated for cell viability using a cell proliferation/cell counting kit-8(CCK-8). The potency of NA and NA-P was evaluated in the safe concentration range, and the effects of 0-100 μmol·L-1 NA and probe on M5-induced proliferation of HaCaT cells were detected using CCK-8. The effects of 20, 40, 80 μmol·L-1 NA and 80 μmol·L-1 NA-P on the expression of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), interleukin-23 (IL-23), and interleukin-17A (IL-17A) inflammatory factors were detected by enzyme-linked immunosorbent assay (ELISA), and real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) was used to measure the effects of NA on the mRNA expression of keratin 16 (K16) in HaCaT cells, S100 calcium-binding protein A9 (S100A9), S100 calcium-binding protein A7 (S100A7), IL-6, IL-17A, and chemokine 1 (CXCL1). In vitro fluorescence labeling and competition experiments using NA-P were performed, and target protein angling and analysis using pull-down experiments combined with liquid chromatography-mass spectrometry (Pull-down/LC-MS/MS) were conducted. Target validation was performed using pull-down experiments combined with protein immunoblotting (Pull down-WB), cellular heat transfer analysis combined with protein immunoblot (CETSA-WB) experiments, and molecular docking. Finally, Real-time PCR was utilized to detect the effects of 20, 40, 80 μmol·L-1 NA and 80 μmol·L-1 NA-P on the mRNA expression of IL-1β, nucleotide-binding oligomeric structural domain-like receptor protein 3 (NLRP3), apoptosis-associated speckled-like protein (ASC), and cysteine protease-1 (Caspase-1) in HaCaT cells. Protein immunoblot (Western blot) was used to detect the effects of phosphorylated p65 (p-p65), p65, phosphorylated human nuclear factor-κB inhibitory protein α (p-IκBα), human nuclear factor κB inhibitory protein α (IκBα), and heat shock protein 90 (HSP90) expression. ResultsIn the 200 μmol·L-1 safe concentration range, HaCaT cell proliferation, increased expression of TNF-α, IL-1β, IL-23, and IL-17A inflammatory factors, and increased mRNA expression of K16, S100A9, S100A7, IL-6, IL-17A, and CXCL1 were observed in the M5 group compared with the blank group. Cell proliferation in 5-100 μmol·L-1 NA and NA-P groups was inhibited, and the expression of TNF-α, IL-1β, IL-23, and IL-17A inflammatory factors was decreased in the NA-L, NA-M, NA-H, and NA-P-H groups. The mRNA expression of K16, S100A9, S100A7, IL-6, IL-17A, and CXCL1 was decreased (P<0.05). High-confidence targets were screened for HSP90 protein by Pull-down/LC-MS/MS using 200 μmol·L-1 NA competing with 100 μmol·L-1 NA-P. Compared with that in the blank group, the mRNA expression of NLRP3, IL-1β, ASC, and Caspase-1, as well as the expression of p-p65/p65, p-IκBα/IκBα, and HSP90 protein, were increased in HaCaT cells in the M5 group (P<0.05). Compared with that in the M5 group, the mRNA expression of NLRP3, IL-1β, ASC, and Caspase-1 of cells in the NA-L group, the NA-M group, the NA-H group, and the NA-P-H group was decreased (P<0.05). p-p65/p65 and p-IκBα/IκBα were decreased in the NA-M and NA-H groups (P<0.05), and there was no change in HSP90 protein. Pull down-WB showed that NA could directly target HSP90 protein, and NA binding to HSP90 protein enhanced its thermal stability. Molecular docking of NA with HSP90 family proteins HSP90AA1, HSP90B1, and HSP90AB1 all resulted in highly stable binding. ConclusionNA can inhibit the proliferation and inflammatory response of psoriatic keratinocytes by a mechanism that may be achieved by targeting HSP90 to modulate the NF-κB/NLRP3 signaling pathway.
3.Therapeutic Effect of Cranial Painkiller Pills' Extract Powder in Treatment of Trigeminal Neuralgia Induced by Injection of Talci Pulvis into Infraorbital Foramen of Model Rats Based on OTULIN-regulated Neuroinflammation
Shuran LI ; Xinwei WANG ; Jing SUN ; Dan XIE ; Ronghua ZHAO ; Lei BAO ; Zihan GENG ; Qiyue SUN ; Jingsheng ZHANG ; Yaxin WANG ; Xihe CUI ; Xinying LI ; Bing HAN ; Tianjiao LU ; Xiaolan CUI ; Liying LIU ; Shanshan GUO
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(21):21-28
ObjectiveThis paper aims to verify the therapeutic effect of Cranial Painkiller pills' extract powder prepared by the new process on the rat's trigeminal neuralgia model caused by infraorbital injection of Talci Pulvis, evaluate its potential clinical application value, and compare the therapeutic effect with that of Cranial Painkiller granules, so as to provide data support for the application of the Cranial Painkiller pills' extract powder and precise treatment. MethodsThe rat's trigeminal neuralgia model was constructed by infraorbital injection of Talci Pulvis, and the rats were randomly divided into the normal group, model group, carbamazepine group (60 mg·kg-1), Cranial Painkiller granules group (2.70 g·kg-1), and low, medium, and high dosage groups of Cranial Painkiller pills' extract powder (1.35, 2.70, 5.40 g·kg-1) according to the basal mechanical pain thresholds, and there were 10 rats in each group. The drug was administered by gavage to each group 2 h after modeling, and distilled water was given by gavage to the normal and model groups under the same conditions once a day for 10 d. Von Frey brushes were used to measure mechanical pain thresholds in rats. Hematoxylin-eosin (HE) staining was used to detect pathological changes in the trigeminal ganglion, and enzyme-linked immunosorbent assay (ELISA) was used to detect the inflammatory factors interleukin-1 (IL-1), interleukin-6 (IL-6), interleukin-8 (IL-8), and tumor necrosis factor-α (TNF-α) levels in rat serum, as well as neuropeptide substance P (SP) and β-endorphin (β-EP) levels in rat brain tissue. Western blot technique was used to detect the levels of NLRP3, ASC, Caspase-1, and OTULIN proteins in rat brain tissue. ResultsCompared with the normal group, the pain threshold of rats in the model group showed a continuous significant decrease (P<0.01). The pathological damage of brain tissue was significant (P<0.01), and the inflammatory levels of IL-1, IL-6, IL-8, and TNF-α in serum were significantly elevated (P<0.01). The level of the SP in the brain tissue was significantly elevated (P<0.01), and the level of β-EP was significantly reduced (P<0.01), while the level of OTULIN was significantly reduced, and NLRP3, ASC, and Caspase-1 protein levels were significantly elevated (P<0.01). After administration of the drug, compared with the model group, the pain threshold of each dose group of the Cranial Painkiller pills' extract powder and the Cranial Painkiller granules group significantly increased (P<0.01). The inflammatory levels of IL-1, IL-6, IL-8, and TNF-α and SP levels significantly decreased (P<0.01), and the β-EP levels were significantly elevated (P<0.01), while the levels of OTULIN protein were significantly elevated (P<0.05, P<0.01), and the levels of NLRP3, ASC proteins were decreased (P<0.01)in high dose Cranial Painkiller pills' extract powder. Meanwhile, compared with those in the model group, the trigeminal ganglion lesions of rats in the Cranial Painkiller pills' extract powder and Cranial Painkiller granules groups showed different degrees of improvement (P<0.05, P<0.01). ConclusionThe Cranial Painkiller pills' extract powder has significant therapeutic effects on the rat model of trigeminal neuralgia induced by infraorbital injection of Talci Pulvis, and its mechanism is related to the improvement of OTULIN-regulated neuroinflammation.
4.Therapeutic Effect of Jidesheng Sheyao Tablets on VZV-induced Skin Vesicles in Guinea Pigs and Postherpetic Neuralgia in Rats
Rui XIE ; Shanshan GUO ; Zihan GENG ; Lei BAO ; Ronghua ZHAO ; Shuran LI ; Qiyue SUN ; Xinwei WANG ; Jingsheng ZHANG ; Xihe CUI ; Xiaolan CUI ; Jing SUN
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(21):38-47
ObjectiveThis paper aims to investigate the therapeutic effects of Jidesheng Sheyao tablets on varicella-zoster virus (VZV) and its associated postherpetic neuralgia (PHN) to provide experimental evidence for the clinical application and secondary development of Jidesheng Sheyao tablets. MethodsFifty-six guinea pigs were randomly divided into seven groups according to their body weight, namely the normal group, the model group, the positive control group, the high-dose group, medium-dose group, and low-dose group of Jidesheng Sheyao tablets (1.92, 0.96, 0.48 g·d-1), and the group treated with oral administration combined with topical application of Jidesheng Sheyao tablets (0.96 g·d-1 + 1.2 g·kg-1·d-1). The skin on the back of the guinea pigs in each group was depilated and then abraded with sandpaper. Except for the normal group, 200 μL of VZV solution was dropped on the damaged parts of the back of the guinea pigs in the other groups, and the infection lasted for 2 consecutive days. The drug administration started 2 hours after the infection on the first day and lasted for 7 days. The pathological changes of the back of the guinea pigs in each group were observed every day starting from the second day after the infection. On the 7th day, the guinea pigs were sacrificed by CO2 anesthesia. The locally infected skin was taken, and the viral DNA nucleic acid load was detected by real-time polymerase chain reaction (Real-time PCR). The pathological histology examination was carried out after hematoxylin-eosin (HE) staining. Seventy rats were randomly divided into seven groups according to their body weight, namely the normal group, the model group, the positive control group, the high-dose group, medium-dose group, and low-dose group of Jidesheng Sheyao tablets (1.08, 0.54, 0.27 g·d-1), and the group treated with oral administration combined with topical application of Jidesheng Sheyao tablets (0.54 g·d-1 + 1.2 g·kg-1·d-1). The rats in each group (except the normal group) were subcutaneously inoculated with 50 μL of VZV suspension between the web of the first and second fingers of the left forelimb. The skin on the back of the rats was depilated, and the drug administration started 2 hours after the infection and lasted for 10 days. The mechanical withdrawal threshold of the paws of the rats was detected by a Von Frey filament algometer before inoculation and on the 1st, 3rd, 6th, 8th, and 10th days after inoculation, and the thermal withdrawal reflex latency of the paws of the rats was detected by a hot and cold plate algometer. On the 10th day after the virus inoculation, the rats were anesthetized after the behavioral examination, and the dorsal root ganglia of the spinal cord and spinal cord segments were taken. The contents of substance P (SP), neurokinin (NK), tumor necrosis factor-α (TNF-α), and interleukin-1β (IL-1β) in the dorsal root ganglia of the spinal cord and spinal cord were detected by enzyme-linked immunosorbent assay (ELISA). ResultsCompared with those in the normal group, the guinea pigs in the model group had obvious skin herpes lesions (P<0.01). The viral nucleic acid load was high (P<0.01), and there were disorganized subcutaneous cellular structures and obvious infiltration of inflammatory cells and cell necrosis (P<0.01). The mechanical withdrawal threshold of the paws and the thermal withdrawal reflex latency of the paws of the rats were significantly decreased (P<0.05), and the contents of NK, SP, TNF-α, and IL-1β in the dorsal root ganglia of the spinal cord and spinal cord of the rats were significantly increased (P<0.01). Compared with the model group, the high-dose and medium-dose groups of topical administration of Jidesheng Sheyao tablets and the group of oral administration combined with topical application could significantly improve the lesions such as skin redness and herpes of the guinea pigs caused by VZV infection (P<0.01), reduce the VZV viral nucleic acid load in the skin tissues of the guinea pigs (P<0.01), alleviate the degree of inflammatory cell infiltration and skin cell necrosis in the skin tissue (P<0.05), significantly increase the mechanical withdrawal threshold of the paws and the thermal withdrawal reflex latency of the paws of the rats (P<0.05), and decrease the contents of NK, SP, TNF-α, and IL-1β in the dorsal root ganglia of the spinal cord and spinal cord of the rats (P<0.01). ConclusionJidesheng Sheyao tablets demonstrated significant therapeutic effects on VZV-induced skin infections and postherpetic neuralgia (PHN), providing a promising candidate for the prevention and treatment of VZV infections.
5.Mechanism of Tibetan Medicine Sanwei Doukoutang to Improve Cognitive Dysfunction in 5×FAD Mice Based on Wnt/β-catenin Signaling Pathway
Shuran LI ; Yaxin WANG ; Jing SUN ; Lei BAO ; Zihan GENG ; Dan XIE ; Ronghua ZHAO ; Yanyan BAO ; Qiyue SUN ; Jingsheng ZHANG ; Xinwei WANG ; Xinying LI ; Xihe CUI ; Xiaowei YANG ; LIUXIAN ; Mengyao CUI ; Qingshan LIU ; Shanshan GUO
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(21):54-60
ObjectiveTo explore the effects of the Tibetan medicine Sanwei Doukoutang (SWDK) on cognitive dysfunction in mice suffering from Alzheimer's disease (AD) and its related mechanism. MethodsFifty SPF 5 × FAD mice were randomly divided into model group, total ginsenoside group(0.04 g·kg-1), high-, medium-, and low-dose groups of SWDK (32.60, 16.30, 8.15 g·kg-1), with 10 mice in each group, and ten wild-type mice of the same age were used as the normal group, male and female in 1∶1. Gavage administration was performed once daily for 8 weeks. The Morris water maze test and contextual fear memory experiment were used to observe learning and memory function. Hematoxylin-eosin (HE) staining was utilized to observe the changes in the pathomorphology of brain tissue in mice. The levels of synaptophysin (SYP) and postsynaptic dense substance 95 (PSD95) in mice serum were detected by enzyme-linked immunosorbent assay (ELISA). The positive expression of brain-derived neurotrophic factor(BDNF) in the dentate gyrus (DG) region of mouse brain tissue was observed by immunohistochemistry (IHC). The protein levels of BDNF, Wnt family member 3A(Wnt3a), and β-catenin were detected in the hippocampus of mice by Western blot. ResultsCompared with the normal group of mice, the model group of mice had significantly more complex swimming routes and lower swimming speed (P<0.01), significantly lower percentage of time spent in the target quadrant (P<0.01), and a significantly lower percentage of freezing time (P<0.05). The number of neurons in the hippocampal region of mice was obviously reduced and unevenly arranged. The levels of SYP and PSD95(P<0.01) in the serum of mice were reduced, and the positive expression of BDNF in the DG region of the brain tissue of mice was reduced. The levels of hippocampal BDNF, Wnt3a, and β-catenin proteins in the hippocampus of mice were obviously reduced (P<0.05, P<0.01). Compared with the model group, the mice in the SWDK group and the total ginsenoside group had significantly shorter swimming routes, the high- and medium- dose SWDK groups significantly higher swimming speeds (P<0.01), significantly higher percentage of time spent in the target quadrant (P<0.01), obviously higher percentage of Freezing time (P<0.05), and obviously more neurons in the hippocampal region of the mice with tighter arrangement. The mice had elevated levels of serum SYP (P<0.05, P<0.01), PSD95 (P<0.01), increased BDNF-positive cells in the DG region of brain tissue, and obviously elevated levels of BDNF, Wnt3a, and β-catenin proteins in the hippocampus of mice (P<0.05, P<0.01). ConclusionSWDK can significantly improve the cognitive dysfunction of AD mice, and its mechanism may be related to regulating the Wnt/β-catenin signaling pathway, which promotes BDNF expression and thereby enhances synaptic plasticity, allowing neuronal signaling to be restored.
6.Therapeutic Effect of Cranial Painkiller Pills' Extract Powder in Treatment of Trigeminal Neuralgia Induced by Injection of Talci Pulvis into Infraorbital Foramen of Model Rats Based on OTULIN-regulated Neuroinflammation
Shuran LI ; Xinwei WANG ; Jing SUN ; Dan XIE ; Ronghua ZHAO ; Lei BAO ; Zihan GENG ; Qiyue SUN ; Jingsheng ZHANG ; Yaxin WANG ; Xihe CUI ; Xinying LI ; Bing HAN ; Tianjiao LU ; Xiaolan CUI ; Liying LIU ; Shanshan GUO
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(21):21-28
ObjectiveThis paper aims to verify the therapeutic effect of Cranial Painkiller pills' extract powder prepared by the new process on the rat's trigeminal neuralgia model caused by infraorbital injection of Talci Pulvis, evaluate its potential clinical application value, and compare the therapeutic effect with that of Cranial Painkiller granules, so as to provide data support for the application of the Cranial Painkiller pills' extract powder and precise treatment. MethodsThe rat's trigeminal neuralgia model was constructed by infraorbital injection of Talci Pulvis, and the rats were randomly divided into the normal group, model group, carbamazepine group (60 mg·kg-1), Cranial Painkiller granules group (2.70 g·kg-1), and low, medium, and high dosage groups of Cranial Painkiller pills' extract powder (1.35, 2.70, 5.40 g·kg-1) according to the basal mechanical pain thresholds, and there were 10 rats in each group. The drug was administered by gavage to each group 2 h after modeling, and distilled water was given by gavage to the normal and model groups under the same conditions once a day for 10 d. Von Frey brushes were used to measure mechanical pain thresholds in rats. Hematoxylin-eosin (HE) staining was used to detect pathological changes in the trigeminal ganglion, and enzyme-linked immunosorbent assay (ELISA) was used to detect the inflammatory factors interleukin-1 (IL-1), interleukin-6 (IL-6), interleukin-8 (IL-8), and tumor necrosis factor-α (TNF-α) levels in rat serum, as well as neuropeptide substance P (SP) and β-endorphin (β-EP) levels in rat brain tissue. Western blot technique was used to detect the levels of NLRP3, ASC, Caspase-1, and OTULIN proteins in rat brain tissue. ResultsCompared with the normal group, the pain threshold of rats in the model group showed a continuous significant decrease (P<0.01). The pathological damage of brain tissue was significant (P<0.01), and the inflammatory levels of IL-1, IL-6, IL-8, and TNF-α in serum were significantly elevated (P<0.01). The level of the SP in the brain tissue was significantly elevated (P<0.01), and the level of β-EP was significantly reduced (P<0.01), while the level of OTULIN was significantly reduced, and NLRP3, ASC, and Caspase-1 protein levels were significantly elevated (P<0.01). After administration of the drug, compared with the model group, the pain threshold of each dose group of the Cranial Painkiller pills' extract powder and the Cranial Painkiller granules group significantly increased (P<0.01). The inflammatory levels of IL-1, IL-6, IL-8, and TNF-α and SP levels significantly decreased (P<0.01), and the β-EP levels were significantly elevated (P<0.01), while the levels of OTULIN protein were significantly elevated (P<0.05, P<0.01), and the levels of NLRP3, ASC proteins were decreased (P<0.01)in high dose Cranial Painkiller pills' extract powder. Meanwhile, compared with those in the model group, the trigeminal ganglion lesions of rats in the Cranial Painkiller pills' extract powder and Cranial Painkiller granules groups showed different degrees of improvement (P<0.05, P<0.01). ConclusionThe Cranial Painkiller pills' extract powder has significant therapeutic effects on the rat model of trigeminal neuralgia induced by infraorbital injection of Talci Pulvis, and its mechanism is related to the improvement of OTULIN-regulated neuroinflammation.
7.Therapeutic Effect of Jidesheng Sheyao Tablets on VZV-induced Skin Vesicles in Guinea Pigs and Postherpetic Neuralgia in Rats
Rui XIE ; Shanshan GUO ; Zihan GENG ; Lei BAO ; Ronghua ZHAO ; Shuran LI ; Qiyue SUN ; Xinwei WANG ; Jingsheng ZHANG ; Xihe CUI ; Xiaolan CUI ; Jing SUN
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(21):38-47
ObjectiveThis paper aims to investigate the therapeutic effects of Jidesheng Sheyao tablets on varicella-zoster virus (VZV) and its associated postherpetic neuralgia (PHN) to provide experimental evidence for the clinical application and secondary development of Jidesheng Sheyao tablets. MethodsFifty-six guinea pigs were randomly divided into seven groups according to their body weight, namely the normal group, the model group, the positive control group, the high-dose group, medium-dose group, and low-dose group of Jidesheng Sheyao tablets (1.92, 0.96, 0.48 g·d-1), and the group treated with oral administration combined with topical application of Jidesheng Sheyao tablets (0.96 g·d-1 + 1.2 g·kg-1·d-1). The skin on the back of the guinea pigs in each group was depilated and then abraded with sandpaper. Except for the normal group, 200 μL of VZV solution was dropped on the damaged parts of the back of the guinea pigs in the other groups, and the infection lasted for 2 consecutive days. The drug administration started 2 hours after the infection on the first day and lasted for 7 days. The pathological changes of the back of the guinea pigs in each group were observed every day starting from the second day after the infection. On the 7th day, the guinea pigs were sacrificed by CO2 anesthesia. The locally infected skin was taken, and the viral DNA nucleic acid load was detected by real-time polymerase chain reaction (Real-time PCR). The pathological histology examination was carried out after hematoxylin-eosin (HE) staining. Seventy rats were randomly divided into seven groups according to their body weight, namely the normal group, the model group, the positive control group, the high-dose group, medium-dose group, and low-dose group of Jidesheng Sheyao tablets (1.08, 0.54, 0.27 g·d-1), and the group treated with oral administration combined with topical application of Jidesheng Sheyao tablets (0.54 g·d-1 + 1.2 g·kg-1·d-1). The rats in each group (except the normal group) were subcutaneously inoculated with 50 μL of VZV suspension between the web of the first and second fingers of the left forelimb. The skin on the back of the rats was depilated, and the drug administration started 2 hours after the infection and lasted for 10 days. The mechanical withdrawal threshold of the paws of the rats was detected by a Von Frey filament algometer before inoculation and on the 1st, 3rd, 6th, 8th, and 10th days after inoculation, and the thermal withdrawal reflex latency of the paws of the rats was detected by a hot and cold plate algometer. On the 10th day after the virus inoculation, the rats were anesthetized after the behavioral examination, and the dorsal root ganglia of the spinal cord and spinal cord segments were taken. The contents of substance P (SP), neurokinin (NK), tumor necrosis factor-α (TNF-α), and interleukin-1β (IL-1β) in the dorsal root ganglia of the spinal cord and spinal cord were detected by enzyme-linked immunosorbent assay (ELISA). ResultsCompared with those in the normal group, the guinea pigs in the model group had obvious skin herpes lesions (P<0.01). The viral nucleic acid load was high (P<0.01), and there were disorganized subcutaneous cellular structures and obvious infiltration of inflammatory cells and cell necrosis (P<0.01). The mechanical withdrawal threshold of the paws and the thermal withdrawal reflex latency of the paws of the rats were significantly decreased (P<0.05), and the contents of NK, SP, TNF-α, and IL-1β in the dorsal root ganglia of the spinal cord and spinal cord of the rats were significantly increased (P<0.01). Compared with the model group, the high-dose and medium-dose groups of topical administration of Jidesheng Sheyao tablets and the group of oral administration combined with topical application could significantly improve the lesions such as skin redness and herpes of the guinea pigs caused by VZV infection (P<0.01), reduce the VZV viral nucleic acid load in the skin tissues of the guinea pigs (P<0.01), alleviate the degree of inflammatory cell infiltration and skin cell necrosis in the skin tissue (P<0.05), significantly increase the mechanical withdrawal threshold of the paws and the thermal withdrawal reflex latency of the paws of the rats (P<0.05), and decrease the contents of NK, SP, TNF-α, and IL-1β in the dorsal root ganglia of the spinal cord and spinal cord of the rats (P<0.01). ConclusionJidesheng Sheyao tablets demonstrated significant therapeutic effects on VZV-induced skin infections and postherpetic neuralgia (PHN), providing a promising candidate for the prevention and treatment of VZV infections.
8.Mechanism of Tibetan Medicine Sanwei Doukoutang to Improve Cognitive Dysfunction in 5×FAD Mice Based on Wnt/β-catenin Signaling Pathway
Shuran LI ; Yaxin WANG ; Jing SUN ; Lei BAO ; Zihan GENG ; Dan XIE ; Ronghua ZHAO ; Yanyan BAO ; Qiyue SUN ; Jingsheng ZHANG ; Xinwei WANG ; Xinying LI ; Xihe CUI ; Xiaowei YANG ; LIUXIAN ; Mengyao CUI ; Qingshan LIU ; Shanshan GUO
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(21):54-60
ObjectiveTo explore the effects of the Tibetan medicine Sanwei Doukoutang (SWDK) on cognitive dysfunction in mice suffering from Alzheimer's disease (AD) and its related mechanism. MethodsFifty SPF 5 × FAD mice were randomly divided into model group, total ginsenoside group(0.04 g·kg-1), high-, medium-, and low-dose groups of SWDK (32.60, 16.30, 8.15 g·kg-1), with 10 mice in each group, and ten wild-type mice of the same age were used as the normal group, male and female in 1∶1. Gavage administration was performed once daily for 8 weeks. The Morris water maze test and contextual fear memory experiment were used to observe learning and memory function. Hematoxylin-eosin (HE) staining was utilized to observe the changes in the pathomorphology of brain tissue in mice. The levels of synaptophysin (SYP) and postsynaptic dense substance 95 (PSD95) in mice serum were detected by enzyme-linked immunosorbent assay (ELISA). The positive expression of brain-derived neurotrophic factor(BDNF) in the dentate gyrus (DG) region of mouse brain tissue was observed by immunohistochemistry (IHC). The protein levels of BDNF, Wnt family member 3A(Wnt3a), and β-catenin were detected in the hippocampus of mice by Western blot. ResultsCompared with the normal group of mice, the model group of mice had significantly more complex swimming routes and lower swimming speed (P<0.01), significantly lower percentage of time spent in the target quadrant (P<0.01), and a significantly lower percentage of freezing time (P<0.05). The number of neurons in the hippocampal region of mice was obviously reduced and unevenly arranged. The levels of SYP and PSD95(P<0.01) in the serum of mice were reduced, and the positive expression of BDNF in the DG region of the brain tissue of mice was reduced. The levels of hippocampal BDNF, Wnt3a, and β-catenin proteins in the hippocampus of mice were obviously reduced (P<0.05, P<0.01). Compared with the model group, the mice in the SWDK group and the total ginsenoside group had significantly shorter swimming routes, the high- and medium- dose SWDK groups significantly higher swimming speeds (P<0.01), significantly higher percentage of time spent in the target quadrant (P<0.01), obviously higher percentage of Freezing time (P<0.05), and obviously more neurons in the hippocampal region of the mice with tighter arrangement. The mice had elevated levels of serum SYP (P<0.05, P<0.01), PSD95 (P<0.01), increased BDNF-positive cells in the DG region of brain tissue, and obviously elevated levels of BDNF, Wnt3a, and β-catenin proteins in the hippocampus of mice (P<0.05, P<0.01). ConclusionSWDK can significantly improve the cognitive dysfunction of AD mice, and its mechanism may be related to regulating the Wnt/β-catenin signaling pathway, which promotes BDNF expression and thereby enhances synaptic plasticity, allowing neuronal signaling to be restored.
9.Dynamic disinfection effect of the upper-room 222 nm ultraviolet radiation on the air in the emergency department in a tertiary general hospital in Beijing, China
Shi CHENG ; Bin XU ; Yue DU ; Jing LI ; Yingxin MA ; Xiaojuan MENG ; Wei HAN ; Xinwei YU ; Aixiang HU ; Yuewei ZHANG
Shanghai Journal of Preventive Medicine 2024;36(9):836-841
ObjectiveTo evaluate the dynamic disinfection effect of the upper-room 222 nm ultraviolet radiation on the air in different areas of the emergency department, and to provide references for a new solution of air disinfection that man-machine coexisted in the medical and healthcare institutions. MethodsThe upper-room 222 nm ultraviolet radiation air sterilizers were installed at a height of 2.3‒2.6 m from the ground in the observation room, computed tomography (CT) scanning room, rescue room and consulting room of the emergency department in Beijing Tiantan Hospital, Capital Medical University . The test area was divided into a 222 nm ultraviolet group and a control group according to whether the 222 nm ultraviolet germicidal air disinfection was conducted or not. The experimental group started air disinfection at8:00 a.m., and the air sampling was conducted from 9:00 a.m. to 16:00 p.m., with a 10 min sampling interval of every 1 hour. While the control group only collected air sample with the same air sampling method used in the experimental group, without air disinfection. The air microbial sampler with six-level sieve impingement was used for the air sampling, and the differences in the total number of airborne bacterial colonies were compared between the two groups. ResultsA total of 128 air samples were collected in the trial, of which 64 were from the experimental group and 64 from the control group. The total number of airborne bacterial colonies in the experimental group was lower than that in the control group (all P<0.001), and was maintained at a lower level throughout the entire period. The killing rate of 222 nm ultraviolet radiation to airborne microorganisms was approximately 55.76% to 76.33% in different sampling areas. The qualified rates of the total number of dynamic airborne bacterial colonies in the observation room, rescue room and consulting room were improved from 12.50%, 37.50% and 25.00% to 81.25%, 100.00% and 100.00%, respectively (all P<0.001). Over 62.50% of the time, the air quality in the consulting room and CT room in the 222 nm ultraviolet group met the environment standards for airborne bacterial colony criteria of class Ⅰ or class Ⅱ. ConclusionThe upper-room222 nm ultraviolet radiation germicidal disinfection can effectively reduce the total number of airborne bacterial colonies and improve the environment for emergency department, and the continuous using of it is helpful for keeping the air safe and clean.
10.Relationship between residential neighborhood green space and mental health of middle school students
ZHANG Shunjun, RAN Fei, WU Wenqiao, ZENG Qingwen, YUE Xinwei
Chinese Journal of School Health 2024;45(10):1426-1430
Objective:
To understand the relationship between residential neighborhood green space and mental health of middle school students, so as to provide a theoretical basis for mental health promotion among of middle school students at the level of residential environment.
Methods:
From October to December 2023, 1 258 students from three junior high schools and two general high schools in Panlong, Wuhua, Chenggong, and Guandu districts of Kunming City were selected by stratified cluster random sampling method. Conducting onsite questionnaire survey using the Mental Health Inventroy of Middle School students, residential neighborhood green space were evaluated using normalized difference vegetation index (NDVI). Analysis of variance, Pearson correlation analysis and linear regression analysis were used to analyze the relationship between as green space in residential neighborhoods and the mental health of middle school students.
Results:
The average mental health score of middle school students in Kunming was (2.02± 0.69 ), of which 447 (37.92%) had mild mental health problems, 107 (8.51%) had moderate mental health problems, and 12 ( 0.95% ) had more severe mental health problems. Mental health scores of middle school students varied significantly by gender, grade, and family income ( t/F =6.99, 7.15, 3.70, P <0.05). Correlation analysis showed that NDVI_250 m, NDVI_500 m, and NDVI_1 000 m were all negatively correlated with the total mental health scores of middle school students ( r =-0.07,-0.09, -0.10 , P <0.05). Multiple linear regression analysis showed that gender, grade, NDVI_250 m, NDVI_500 m, and NDVI_1 000 m were infuencing factors on the mental health of middle school students( B =0.25,0.05,-0.47,-0.67,-0.78, P <0.05).
Conclusions
Mental health of middle school students in Kunming is related to green space in different buffer zones around residences. The mental health education of middle school students should be strengthened, especially for girls and senior students, optimizing resources to reduce pressure, and improving the quality of green space in residential communities to improve the mental health of adolescents.


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