1.Effect of Xiaoqinglong Decoction (小青龙汤) on Th2 Immune Responses and Nasal Mucosal TSLP/OX40L Pathway in Allergic Rhinitis Model Rats
Yimeng CHEN ; Yuye CHEN ; Guangchun YU ; Bei CHEN ; Jianwei ZHANG ; Shanshan DING ; Xiaoting YANG ; Baifan YU ; Yating CAI ; Xuejuan LIN ; Mengting ZHANG
Journal of Traditional Chinese Medicine 2026;67(9):994-1002
ObjectiveTo explore the potential mechanism of Xiaoqinglong Decoction (小青龙汤, XD) in the treatment of allergic rhinitis. MethodsForty-five rats were randomly assigned to a control group, a model group, a loratadine group, low-, medium- and high-dose XD groups, and low-, medium- and high-dose Mahuang Decoction and Cang'erzi Powder (麻黄汤合苍耳子散, MDCP) groups. Except for the control group, rats were administered with ovalbumin (OVA) and aluminum hydroxide via intraperitoneal injection for 14 days to establish an allergic rhinitis model. After the 14th-day injection, nasal stimulation was continued with 20 μl of 10% OVA solution to maintain the model. Rats in the control group and the model group received 10 ml/(kg·d) of saline, whereas those in the loratadine group were administered with 0.9 mg/(kg·d) of loratadine. The low-, medium- and high-dose XD groups were administered XD at the dose of 2.7, 5.4, and 10.8 g/(kg·d), respectively. The low-, medium- and high-dose MDCP groups were administered MDCP at the dose of 2.43, 4.86, and 9.72 g/(kg·d), respectively. All treatments were administered by gavage once daily for 7 consecutive days. One hour after the final gavage, nasal symptom scores were recorded for all group of rats. The next day, serum levels of immunoglobulin E (IgE), interleukin-4 (IL-4), and interleukin-13 (IL-13) were measured. HE staining was used to observe the pathological morphology of the nasal mucosal tissue. Quantitative reverse transcription PCR (RT-qPCR) and Western Blot were performed to assess mRNA and protein expression of thymic stromal lymphopoietin (TSLP) and OX40 ligand (OX40L) in the nasal mucosa. ResultsCompared to the control group, total nasal symptom score in the model group significantly increased (P<0.01). HE staining revealed disrupted and adhered cilia, thickened basement membranes, and extensive inflammatory cell infiltration in the nasal mucosa. Serum levels of total IgE, IL-4, and IL-13, as well as TSLP and OX40L mRNA and protein expression in the nasal mucosa, were significantly elevated in the model group (P<0.05 or P<0.01). Compared to the model group, the total nasal symptom scores in all drug intervention groups were significantly reduced; the serum total IgE levels in the loratadine group, the low- and medium-dose XD groups, and the low- and high-dose MDCP groups were significantly reduced; and the serum levels of IL-4 and IL-13 in the high-dose XD group and the high-dose MDCP group decreased (P<0.05 or P<0.01). Nasal mucosal structure was improved. Except for the low-dose MDCP group, all other intervention groups showed a significant reduction in TSLP and OX40L mRNA expression in the nasal mucosa (P<0.01). All doses of XD and the medium- and high-dose MDCP groups significantly decreased the protein levels of TSLP and OX40L (P<0.05). The medium-dose XD group exhibited more improvement of nasal symptom scores and greater suppression of expression of TSLP and OX40L mRNA, and TSLP protein levels compared to the loratadine group (P<0.05). ConclusionXD may protect nasal mucosa of rats and alleviate allergic rhinitis by suppressing the TSLP/OX40L pathway, thereby attenuating Th2-mediated immune responses.
2.Effect of Xiaoqinglong Decoction (小青龙汤) on Th2 Immune Responses and Nasal Mucosal TSLP/OX40L Pathway in Allergic Rhinitis Model Rats
Yimeng CHEN ; Yuye CHEN ; Guangchun YU ; Bei CHEN ; Jianwei ZHANG ; Shanshan DING ; Xiaoting YANG ; Baifan YU ; Yating CAI ; Xuejuan LIN ; Mengting ZHANG
Journal of Traditional Chinese Medicine 2026;67(9):994-1002
ObjectiveTo explore the potential mechanism of Xiaoqinglong Decoction (小青龙汤, XD) in the treatment of allergic rhinitis. MethodsForty-five rats were randomly assigned to a control group, a model group, a loratadine group, low-, medium- and high-dose XD groups, and low-, medium- and high-dose Mahuang Decoction and Cang'erzi Powder (麻黄汤合苍耳子散, MDCP) groups. Except for the control group, rats were administered with ovalbumin (OVA) and aluminum hydroxide via intraperitoneal injection for 14 days to establish an allergic rhinitis model. After the 14th-day injection, nasal stimulation was continued with 20 μl of 10% OVA solution to maintain the model. Rats in the control group and the model group received 10 ml/(kg·d) of saline, whereas those in the loratadine group were administered with 0.9 mg/(kg·d) of loratadine. The low-, medium- and high-dose XD groups were administered XD at the dose of 2.7, 5.4, and 10.8 g/(kg·d), respectively. The low-, medium- and high-dose MDCP groups were administered MDCP at the dose of 2.43, 4.86, and 9.72 g/(kg·d), respectively. All treatments were administered by gavage once daily for 7 consecutive days. One hour after the final gavage, nasal symptom scores were recorded for all group of rats. The next day, serum levels of immunoglobulin E (IgE), interleukin-4 (IL-4), and interleukin-13 (IL-13) were measured. HE staining was used to observe the pathological morphology of the nasal mucosal tissue. Quantitative reverse transcription PCR (RT-qPCR) and Western Blot were performed to assess mRNA and protein expression of thymic stromal lymphopoietin (TSLP) and OX40 ligand (OX40L) in the nasal mucosa. ResultsCompared to the control group, total nasal symptom score in the model group significantly increased (P<0.01). HE staining revealed disrupted and adhered cilia, thickened basement membranes, and extensive inflammatory cell infiltration in the nasal mucosa. Serum levels of total IgE, IL-4, and IL-13, as well as TSLP and OX40L mRNA and protein expression in the nasal mucosa, were significantly elevated in the model group (P<0.05 or P<0.01). Compared to the model group, the total nasal symptom scores in all drug intervention groups were significantly reduced; the serum total IgE levels in the loratadine group, the low- and medium-dose XD groups, and the low- and high-dose MDCP groups were significantly reduced; and the serum levels of IL-4 and IL-13 in the high-dose XD group and the high-dose MDCP group decreased (P<0.05 or P<0.01). Nasal mucosal structure was improved. Except for the low-dose MDCP group, all other intervention groups showed a significant reduction in TSLP and OX40L mRNA expression in the nasal mucosa (P<0.01). All doses of XD and the medium- and high-dose MDCP groups significantly decreased the protein levels of TSLP and OX40L (P<0.05). The medium-dose XD group exhibited more improvement of nasal symptom scores and greater suppression of expression of TSLP and OX40L mRNA, and TSLP protein levels compared to the loratadine group (P<0.05). ConclusionXD may protect nasal mucosa of rats and alleviate allergic rhinitis by suppressing the TSLP/OX40L pathway, thereby attenuating Th2-mediated immune responses.
3.Effect of Electroacupuncture on Metabolism-Inflammation Network in Patients with Simple Obesity:A Randomized Controlled Trial
Qi SHU ; Qiumei PENG ; Zhihao XU ; Shanshan FANG ; Wenjing SHAO ; Hongliang CHENG ; Rilong HUANG ; Qing YU ; Ronglin CAI
Journal of Traditional Chinese Medicine 2026;67(13):1415-1421
ObjectiveTo observe the clinical efficacy of electroacupuncture in treatment of simple obesity patients and its effects on the metabolism-inflammation network. MethodsSeventy-two patients with simple obesity were randomly divided into a electroacupuncture group and a sham electroacupuncture group, with 36 patients in each group. In the electroacupuncture group, conventional acupuncture was applied at Zhongwan (CV 12), bilateral Tianshu (ST 25), bilateral Daheng (SP15), Qihai (CV 6), Guanyuan (CV 4), bilateral Zusanli (ST 36) and Fenglong (ST 40). The sham electroacupuncture group used placebo needle at same acupoints as the electroacupuncture group. Both groups were given sparse-dense waves with a frequency of 2/10 Hz, 30 minutes per session, 3 times a week, for a total of 8 weeks of treatment. The changes of human body parameters including body weight, body mass index (BMI), waist circumference, and body fat percentage (BFP), metabolic factors including serum fasting blood glucose (FBG), triglyceride (TG), total cholesterol (TC), high density lipoprotein (HDL), low density lipoprotein (LDL), leptin, and ghrelin, and inflammatory factors including interleukin-6 (IL-6) and tumor necrosis factor α (TNF-α) were compared before and after treatment. Correlations between changes in leptin, ghrelin and inflammatory factors were further analyzed. ResultsWaist circumference was significantly reduced after treatment in both groups compared with baseline (P<0.05), and the post-treatment waist circumference was lower in the electroacupuncture group than in the sham electroacupuncture group (P<0.05); body weight, BMI and BFP decreased significantly after treatment compared to pretreatment levels in the electroacupuncture group (P<0.05). Serum leptin, IL-6 and TNF-α declined significantly after treatment in both groups versus baseline (P<0.05); the electroacupuncture group had lower post-treatment levels of leptin, IL-6 and TNF-α as well as larger pre-post changes than the sham group (P<0.05); ghrelin increased markedly after treatment in both groups (P<0.05), with higher post-treatment ghrelin and greater pre-post variation observed in the electroacupuncture group (P<0.05). In the electroacupuncture group, changes in ghrelin were negatively correlated with IL-6 and positively correlated with TNF-α (P<0.01); in the sham electroacupuncture group, ghrelin changes were negatively correlated with both IL-6 and TNF-α, whereas leptin changes were positively correlated with the two inflammatory markers (P<0.01). ConclusionElectroacupuncture can effectively improve clinical manifestations, as well as regulate serum metabolic and pro-inflammatory factors in patients with simple obesity, and discrepant correlation patterns of metabolism-inflammation are observed between the two groups.
4.Effect of macrophage lipolysis-stimulated lipoprotein receptor deficiency on a mouse model of metabolic dysfunction-associated steatohepatitis
Hang MENG ; Qianfeng JIA ; Shanshan DU ; Jiashu XU ; Yongfeng GONG ; Cai MA
Journal of Clinical Hepatology 2026;42(9):2093-2105
ObjectiveTo investigate whether macrophage lipolysis-stimulated lipoprotein receptor (LSR) deficiency affects the progression of metabolic dysfunction-associated steatohepatitis (MASH) by activating the Yes-associated protein (YAP) signaling pathway. MethodsMale wild-type (WT) mice and macrophage-specific Lsr knockout (LsrMφKO, KO) mice, with an age of 8 weeks and similar body weights, were selected and fed with a normal diet, a high-fat diet (HFD), or a high-fat high-cholesterol (HFHC) diet for 30 weeks. HE staining and oil red O staining were used to evaluate liver pathological changes; immunofluorescent staining was used to measure the expression levels of F4/80 and α-smooth muscle actin; RT-qPCR and Western blot were used to measure the levels of markers associated with lipid metabolism, inflammation, and fibrosis. Bone marrow transplantation rescue experiments (WT→KO vs KO→KO groups) were conducted to explore the functional role of LSR in macrophages. At the cellular level, hepatic macrophages were isolated from WT and KO mice to measure the expression levels of YAP signaling pathway-related proteins and their downstream target genes, and in addition, these cells were treated in vitro with verteporfin (VP), a specific inhibitor of YAP, to verify the regulatory effect of this pathway on the expression of inflammatory factors. The independent samples t-test was used for comparison of normally distributed measurement data between two groups, and the Mann-Whitney U test was used for comparison of non-normally distributed measurement data. ResultsUnder the condition of normal diet, there were no significant differences in hepatic indicators between WT and KO mice. KO mice fed an HFD or HFHC diet showed a significant increase in hepatic lipid droplet accumulation, accompanied by lipid metabolism disorder and significant increases in the levels of indicators associated with inflammation and fibrosis (P<0.05). Bone marrow transplantation experiments confirmed that compared with the KO→KO group, the WT→KO group had significant alleviation of liver injury (P<0.05). Mechanistic studies showed that there were significant reductions in the levels of phosphorylated large tumor suppressor kinase 1 (LATS1) and phosphorylated YAP and significant increases in the expression levels of total YAP protein and its downstream target genes in hepatic macrophages of KO mice (P<0.05). In vitro VP intervention significantly inhibited the expression levels of inflammatory factors in hepatic macrophages isolated from KO mice (P<0.05). ConclusionMacrophage LSR deficiency activates the YAP signaling pathway by inhibiting LATS1/2 kinase activity and promotes the expression of inflammatory factors, thereby accelerating the progression of liver injury in a mouse model of MASH.
5.Cluster analysis on Medical Records Coder's Possible Behavior in DRG Payment
Yinghong FU ; Guangying GAO ; Shanshan DAI ; Xinyu CAI ; Jiashuai TIAN ; Jingyi ZHANG ; Liliang ZHANG
Chinese Health Economics 2025;44(2):79-83
Objective:To analysis the possible behavior of coders in the reform of Diagnosis Related Groups(DRG)payment mode.Methods:To collect through questionnaires the possible coders'behaviors,and extract the common factors of possible behaviors according to factor analysis,and cluster the coders by the K-means clustering method.Results:Classify the possible coders'behaviors into positive behaviors such as following coding norms and standards,and negative behaviors such as directly coding without detailed reading of medical records.Less than 25 years old were more inclined to adopt negative behaviors,and associate senior titles and higher cognitive level of DRG were more inclined to adopt positive behaviors.The coders are divided into"positive group"who tend to take positive behaviors and"negative group"who take negative behaviors.Conclusion:Coders are more likely to adopt positive behavior,and we should be pay attention to the impact of different demographic characteristics on coders.To adopt different management strategies for different categories of coders,and strengthen training through hierarchical classification,and enhance coders'cognitive of DRG are more conducive to the selection of positive behaviors.
6.Effects of galectin-3 on oxidative damage and expression of inflammatory fac-tors in retinal pigment epithelial cells induced by high glucose
Pei LIU ; Yu CAI ; Decheng WANG ; Shanshan HAN
Recent Advances in Ophthalmology 2025;45(4):269-275
Objective To investigate the effects of galectin-3(Gal3)on autophagy in high glucose-induced human telomerase reverse transcriptase-immortalized retinal pigment epithelial(hTERT-RPE)cells.Methods The hTERT-RPE cells cultured in vitro were randomly divided into a normal group(group C),a high glucose group(group HG),a high glu-cose+si-NC group(group HG+si-NC)and a high glucose+si-Gal3 group(group HG+si-Gal3).Reverse transcrip-tion polymerase chain reaction(RT-PCR)was used to detect the relative mRNA expression levels of Gal3,intercellular ad-hesion molecule(ICAM)-1,interleukin(IL)-6 and tumor necrosis factor(TNF)-α in hTERT-RPE cells of each group.The expression levels of IL-1 and IL-6 in the supernatant of hTERT-RPE cells were measured by enzyme-linked immunosorbent assay(ELISA).The expression level of reactive oxygen species(ROS),the activity of superoxide dismutase(SOD)and the content of malondialdehyde(MDA)in hTERT-RPE cells were analyzed by dichlorofluorescin diacetate(DCFH-DA)staining,the colorimetric method and the microplate method,respectively.The protein expression levels of Gal3,the ratio of microtubule associated protein 1 light chain 3(LC3)type Ⅱ to type Ⅰ(LC3 Ⅱ/LC3 Ⅰ),Beclin 1,and autophagy-associat-ed 5(ATG5)and 7(ATG7)in hTERT-RPE cells from each group were detected by Western blot analysis.After the hTERT-RPE cells in each group were transfected with double fluorescent mRFP-GFP-LC3 plasmids alone and with RFP-LAMP1 and GFP-LC3 plasmids jointly,the changes of autophagy flow in hTERT-RPE cells were detected by laser confocal microscopy.Results Compared with the group C,the group HG showed an increase in the expression of Gal3,IL-1,IL-6,TNF-α,ICAM-1,P62,ROS and MDA content(all P<0.05).However,the expression of LC3 Ⅱ/LC3 Ⅰ,Beclin1,ATG7,ATG5,the SOD activity,and the number of red(autolysosomes)and yellow fluorescence spots(autophagosomes)of the double fluo-rescent mRFP-GFP-LC3 plasmid were all lower in the group HG than those in the group C(all P<0.05).There was no sig-nificant difference in the number of yellow fluorescent spots(autolysosomes)co-located by RFP-LAMP1 and GFP-LC3 plas-mids between groups C and HG(P>0.05).There were no significant differences in the expression levels of the above-mentioned indexes in hTERT-RPE cells between groups HG and HG+si-NC(all P>0.05).The expression levels of Gal3,IL-1,IL-6,TNF-α,ICAM-1,P62,ROS and MDA content in hTERT-RPE cells of the group HG+si-Gal3 were lower than those of the group HG+si-NC(all P<0.05).Compared with those in the group HG+si-NC,the expression levels of LC3 Ⅱ/LC3 Ⅰ,Beclin1,ATG7,ATG5,SOD activity,and the number of red and yellow fluorescent spots of the double fluo-rescent mRFP-GFP-LC3 plasmid were increased in the group HG+si-Gal3(all P<0.05).There was no significant differ-ence in the number of yellow fluorescent spots co-located by RFP-LAMP1 and GFP-LC3 plasmids between groups HG+si-GaL3 and HG+si-NC(P>0.05).Conclusion Gal3 is significantly elevated in hTERT-RPE cells induced by high glu-cose,resulting in impaired autophagy flow.It produces a direct regulatory effect on the formation of autophagosomes,and plays a highly active role in oxidative damage and expression of inflammatory factors in hTERT-RPE cells induced by high glucose.
7.Exploring the impact of a new family ward model on the language,social,and self-care abilities of chil-dren with autism spectrum language
Shanshan ZHU ; Mingxuan CAI ; Ping ZHOU
Chinese Journal of Rehabilitation Medicine 2025;40(10):1541-1546
Objective:To evaluate the application value of a new family ward model in rehabilitation training for chil-dren with autism spectrum disorder(ASD)and its impact on improving the prognosis..Method:Seventy-two children with ASD admitted to the Children's Health Department of Guanggu Branch of Hubei Maternal and Child Health Hospital from January 2022 to January 2024 were assigned to the study group.Another 66 children with ASD who underwent rehabilitation treatment at the outpatient training center during the same period were included as the control group.All children received comprehensive training meth-ods such as language training,social training,sensory integration training,fine motor training,occupational therapy,behavior correction training,and parent training.Based on the new family ward model,the research group provides family support services for pediatric patients,providing real-time guidance on family rehabilita-tion training,natural scenario generalization,and life skills improvement.All patients are trained 6 hours a day for 6 months.Both groups of children were evaluated by the VB-MAPP,ABC scale,CARS scale,SRS scale,and S-M scale evaluation before and after training for efficacy analysis.Result:After 6 months of training,the scores of CARS and SRS scales in the study group were significantly lower than those in the control group(t=2.89,3.04,P<0.05).Compared with the control group,the VB-MAPP and S-M scale evaluation scores in the study group were significantly improved(t=-2.42,-2.45,P<0.05).Conclusion:The new family ward model can effectively improve the language,daily living skills,and social adaptation abilities of children with ASD,and provide sufficient family support services,thereby improving the prognosis of ASD children more effectively.
8.Exploring the impact of a new family ward model on the language,social,and self-care abilities of chil-dren with autism spectrum language
Shanshan ZHU ; Mingxuan CAI ; Ping ZHOU
Chinese Journal of Rehabilitation Medicine 2025;40(10):1541-1546
Objective:To evaluate the application value of a new family ward model in rehabilitation training for chil-dren with autism spectrum disorder(ASD)and its impact on improving the prognosis..Method:Seventy-two children with ASD admitted to the Children's Health Department of Guanggu Branch of Hubei Maternal and Child Health Hospital from January 2022 to January 2024 were assigned to the study group.Another 66 children with ASD who underwent rehabilitation treatment at the outpatient training center during the same period were included as the control group.All children received comprehensive training meth-ods such as language training,social training,sensory integration training,fine motor training,occupational therapy,behavior correction training,and parent training.Based on the new family ward model,the research group provides family support services for pediatric patients,providing real-time guidance on family rehabilita-tion training,natural scenario generalization,and life skills improvement.All patients are trained 6 hours a day for 6 months.Both groups of children were evaluated by the VB-MAPP,ABC scale,CARS scale,SRS scale,and S-M scale evaluation before and after training for efficacy analysis.Result:After 6 months of training,the scores of CARS and SRS scales in the study group were significantly lower than those in the control group(t=2.89,3.04,P<0.05).Compared with the control group,the VB-MAPP and S-M scale evaluation scores in the study group were significantly improved(t=-2.42,-2.45,P<0.05).Conclusion:The new family ward model can effectively improve the language,daily living skills,and social adaptation abilities of children with ASD,and provide sufficient family support services,thereby improving the prognosis of ASD children more effectively.
9.Consistency Evaluation of Processing of Black Panacis Quinquefolii Radix Decoction Pieces Based on "Chromaticity-chemistry-activity"
Yulong YANG ; Shanshan LI ; Yuting YANG ; Ziqi YANG ; Guangzhi CAI ; Liru ZHAO ; Jiyu GONG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(20):195-203
ObjectiveTo assess the quality consistency of black Panacis Quinquefolii Radix(bPQR) decoction pieces prepared by atmospheric and pressurized steaming processes based on chromaticity-chemical composition-vasoactive inhibition. The ultimate goal was to screen the pressurized steaming process yielding quality equivalent to atmospheric steaming, and optimize the processing technology of bPQR. MethodsThe bPQR decoction pieces were prepared using both atmospheric and pressurized steaming processes, and the chromaticity values[lightness value(L*), red/green chromaticity value(a*), yellow/blue chromaticity value(b*), total chromaticity value(E*ab)] were measured. High performance liquid chromatography(HPLC) was employed to establish fingerprint profiles for the decoction pieces, and cluster analysis was conducted on chromaticity values and the common peak areas in fingerprint profiles to elucidate the quality relationships between the decoction pieces processed by different methods. The optimal atmospheric steaming of bPQR decoction pieces was determined through zebrafish angiogenesis inhibition experiments. The contents of ginsenosides Rg1, Re, Rb1, 20(S)-Rg3, Rk1 and Rg5 in the decoction pieces were quantified, and Spearman correlation analysis was employed to investigate the relationship between saponin content, chromaticity, and angiogenesis inhibition activity during the steaming process. By integrating the consistency of chromaticity, saponin components and angiogenesis inhibition activity, pressurized steaming conditions with quality equivalent to the atmospheric pressure method were selected. ResultsCompared with the atmospheric steaming method, pressurized steaming resulted in faster color darkening and higher conversion rates of ginsenosides in bPQR decoction pieces. Moreover, the neovascularization inhibitory activity of bPQR decoction pieces continued to increase with the deepening of processing. Based on the effectiveness and safety, the optimal process for preparing bPQR decoction pieces with neovascularization inhibitory activity was determined to be atmospheric steaming for 21 h. All six ginsenosides tested exhibited strong to extremely strong correlations with both the chromaticity values of the decoction pieces and their neovascularization inhibitory activities. Among them, ginsenosides Rg1, Re and Rb1 exhibited positive correlations with chromaticity values and negative correlations with zebrafish angiogenesis inhibition activity. Conversely, ginsenosides 20(S)-Rg3, Rk1 and Rg5 showed negative correlations with chromaticity values and positive correlations with zebrafish angiogenesis inhibition activity. By integrating chromaticity values, cluster analysis results, as well as the results of activity, it was determined that the quality of bPQR decoction pieces steamed under pressurized conditions of 110 ℃(0.045 MPa) for 5 h and 115 ℃(0.07 MPa) for 3 h was highly consistent with that obtained by atmospheric steaming for 21 h. ConclusionThe preparation of bPQR decoction pieces by pressurized steaming has the advantages of short preparation time, low energy consumption, and rapid saponin conversion rate, making it a viable alternative to atmospheric steaming for preparing bPQR decoction pieces. Meanwhile, the evaluation method based on chromaticity-chemical composition-activity can provide a more scientific and effective explanation of change rules in the quality during traditional Chinese medicine processing, and offer a new model for optimizing processing technology and enhancing quality control.
10.Quality Evaluation of Black Panacis Quinquefolii Radix Based on Neuroprotective Spectrum-effect Relationship
Yuting YANG ; Shuyun LIANG ; Shanshan LI ; Yulong YANG ; Ziqi YANG ; Guangzhi CAI ; Liru ZHAO ; Jiyu GONG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(18):149-156
ObjectiveTo clarify the neuroprotective effect of black Panacis Quinquefolii Radix(PQR) and explore its active ingredients, with the aim of establishing an activity-oriented quality evaluation method. MethodsTransgenic Tg(HuC∶EGFP) zebrafish was used to establish a neuronal injury model by aluminum chloride immersion. Different doses(10, 20 mg·L-1) of PQR and black PQR ethanol extracts were administered. The neuroprotective effects of PQR and black PQR were compared by analyzing the fluorescent area and intensity of zebrafish neurons. Based on ultra-performance liquid chromatography(UPLC), a fingerprint profile of black PQR was established, followed by principal component analysis(PCA) and orthogonal partial least squares-discriminant analysis(OPLS-DA). Differential components were screened using the criteria of variable importance in the projection(VIP) value>1 and P<0.05. The neuroprotective activity of 14 batches of black PQR was assessed, and Spearman correlation analysis was used to identify saponins related to neuroprotective activity, which were then validated. Based on the above results, active marker components were determined, and an UPLC method was established for their quantitation with clear content limits. ResultsPharmacological efficacy results showed that both PQR and black PQR at different doses could significantly improved neuronal damage in zebrafish. At a dose of 20 mg·L-1, black PQR demonstrated superior efficacy(P<0.05). The fingerprint similarities of 14 batches of black PQR were>0.94, with 26 common peaks identified. Through comparison with the reference standards, 8 components were confirmed, including peak 1(ginsenoside Rg1), peak 2(ginsenoside Re), peak 5(ginsenoside Rb1), peak 9(ginsenoside Rd), peak 16[ginsenoside 20(S)-Rg3], peak 17[ginsenoside 20(R)-Rg3], peak 18(ginsenoside Rk1), and peak 19(ginsenoside Rg5). The results of PCA and OPLS-DA indicated that there were differences in saponins among black PQR samples from different origins, and 12 differential components were screened. All 14 batches of black PQR exhibited good protective effects on zebrafish neurons, with Shaanxi-produced black PQR showing superior protective effects compared to the other three production regions. Spearman correlation analysis revealed that a total of 11 components, including ginsenosides 20(S)-Rg3, 20(R)-Rg3, Rk1 and Rg5, showed a significant positive correlation with the neuroprotective effect in zebrafish(P<0.05). The activity validation results indicated that ginsenosides 20(S)-Rg3, 20(R)-Rg3, Rk1 and Rg5 were the primary components responsible for the neuroprotective effects of black PQR. Quantitative analysis showed that the content of ginsenoside 20(S)-Rg3 in 14 batches of black PQR ranged from 0.17% to 0.52%, and the repair rate of neuronal damage ranged from 42.77% to 97.83%. ConclusionBased on the fingerprint and neuronal protective activity, the spectrum-effect related quality control model of black PQR was established, with ginsenoside 20(S)-Rg3 as the quality control index, and the neuronal damage repair rate≥60% as the evaluation standard, the minimum limit of ginsenoside 20(S)-Rg3 in black PQR should be≥0.20%.

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