1.Exploration on Mechanism of Topical Treatment of Allergic Contact Dermatitis in Mice with Portulacae Herba Based on Nrf2/HO-1/NF-κB Signaling Pathway
Xiaoxue WANG ; Guanwei FAN ; Xiang PU ; Zhongzhao ZHANG ; Xia CHEN ; Ying TANG ; Nana WU ; Jiangli LUO ; Xiangyan KONG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(3):115-123
ObjectiveTo investigate the mechanism of topical treatment of allergic contact dermatitis (ACD) mice with Portulacae Herba based on the nuclear factor E2-related factor 2 (Nrf2)/heme oxygenase-1 (HO-1)/nuclear factor-κB (NF-κB) signaling pathway. MethodsA total of 70 6-week-old specific pathogen free (SPF) female Kunming mice were adaptively fed for 1 week and randomly divided into blank group, model group, compound dexamethasone acetate cream group (2.075×10-2 g·g-1), blank matrix cream group, low-dose Portulacae Herba cream group (0.1 g·g-1), high-dose Portulacae Herba cream group (0.2 g·g-1), and Portulacae Herba + inhibitor group (0.2 g·g-1 + 30 mg·kg-1 ML385), with 10 mice in each group. One day before the experiment, the mice were shaved on the neck and back. Except for the blank group, the mice in the other groups were treated with 2,4-dinitrochlorobenzene (DNCB) to establish an ACD model. After respective administration, the skin lesion of the mice was scored, and the histopathological changes of the skin were stained with hematoxylin-eosin (HE). Enzyme-linked immunosorbent assay (ELISA) was used to detect the levels of interleukin-6 (IL-6), interleukin-1β (IL-1β), reactive oxygen species (ROS), superoxide dismutase (SOD) activity, and malondialdehyde (MDA) in serum of mice. The expression of Nrf2/HO-1/NF-κB signaling pathway-related proteins in mouse skin tissue was detected by immunohistochemistry (IHC), Western blot, and real-time fluorescence quantitative polymerase chain reaction (Real-time PCR). ResultsCompared with the blank group, the mice in the model group had an increased skin lesion score (P<0.01), severe pathological damage to skin tissue, increased content of IL-1β, IL-6, ROS, and MDA in their serum (P<0.01), and decreased content of SOD (P<0.01). In addition, the mRNA and protein expression levels of Nrf2, HO-1, and nuclear factor-κB inhibitor α (IκBα) in skin tissue were up-regulated (P<0.01), while the protein expression levels of phosphorylated (p)-IκBα and p-NF-κB p65 and the mRNA expression of NF-κB p65 were down-regulated (P<0.01). Compared with the model group and the blank matrix cream group, the mice treated with Portulacae Herba had a decreased skin lesion score (P<0.01), reduced pathological damage to skin tissue, decreased content of IL-1β, IL-6, ROS, and MDA in their serum (P<0.01), and increased content of SOD (P<0.01). Additionally, the mRNA and protein expression levels of Nrf2, HO-1, and IκBα in skin tissue were down-regulated (P<0.05,P<0.01), and the protein expression levels of p-IκBα and p-NF-κB p65 and the mRNA expression of NF-κB p65 were up-regulated (P<0.05,P<0.01). Compared with the Portulacae Herba + inhibitor group, the high-dose Portulacae Herba cream group had a decreased skin lesion score (P<0.01), alleviated pathological damage to skin tissue, decreased content of IL-1β, IL-6, ROS, and MDA in the serum of mice (P<0.05,P<0.01), and increased content of SOD (P<0.01). The protein expression levels of Nrf2, HO-1, and IκBα and the mRNA expression of Nrf2 and HO-1 in skin tissue were up-regulated (P<0.05,P<0.01), and the protein expression levels of p-IκBα and p-NF-κB p65 and the mRNA expression of NF-κB p65 were down-regulated (P<0.05). ConclusionPortulacae Herba can improve DNCB-induced ACD skin damage in mice by regulating the Nrf2/HO-1/NF-κB signaling pathway.
2.Effects of Portulaca oleracea cream on skin pruritus and barrier function in allergic contact dermatitis mice
Xiaoxue WANG ; Xia CHEN ; Xiang PU ; Guanwei FAN ; Xiangyan KONG ; Ying TANG ; Nana WU ; Jiangli LUO
China Pharmacy 2025;36(11):1352-1357
OBJECTIVE To study the effects and mechanism of Portulaca oleracea cream on skin pruritus and barrier function in allergic contact dermatitis (ACD) mice. METHODS Low-concentration and high-concentration P. oleracea creams were prepared, with the P. oleracea extract solution (1 g/mL, calculated by crude drug) concentrations of 10% and 20%. Sixty BALB/c mice were randomly allocated into blank group, model group, Mometasone furoate cream group (positive control), blank matrix cream group, P. oleracea low-concentration and high-concentration cream groups. Except for blank group, ACD model was induced in each group using 2,4-dinitrochlorobenzene solution. The blank group and model groups received normal saline, while the remaining groups were treated with their respective creams, once a day, at a dose of approximately 0.5 g per application, continuously for 14 days. At 24 h post-final administration, skin lesions of mice were observed and scored; pathological changes of skin tissue were observed; serum levels of immunoglobulin E(IgE) and tumor necrosis factor-α (TNF-α) were quantified. mRNA expression of MAS-related G protein-coupled receptors (including MrgprA3, MrgprC11, and MrgprD) in dorsal root ganglion (DRG) was assessed; while protein expressions of skin barrier function-related proteins Claudin-1 and Occludin in skin tissues were determined. RESULTS Compared with blank group, mice in the model group exhibited severe skin damage, characterized by loss of epidermal architecture, hyperkeratosis of the skin tissue, and the infiltration of a large number of inflammatory cells. The skin injury scores, as well as the serum levels of IgE and TNF-α, and the mRNA expression levels of MrgprA3, MrgprC11, and MrgprD in DRG, were all significantly elevated compared to the blank group (P<0.05 or P<0.01); in contrast, the protein expression levels of Claudin-1 and Occludin in the skin tissue were markedly reduced (P<0.05). Compared with model group, mice in P. oleracea low-concentration and high- concentration cream groups demonstrated significant alleviation of skin damage, as evidenced by reduced epidermal hyperplasia, mitigated spongiosis in the dermis, and decreased infiltration of inflammatory cells; these quantitative indicators were almost significantly reversed (P<0.05 or P<0.01). No significant differences were observed in the aforementioned skin injuries, pathological alterations, or quantitative indicators between the blank matrix cream group and the model group. CONCLUSIONS P. oleracea may ameliorate skin lesions and restore skin barrier function of ACD mice, the mechanism of which may be associated with downregulating mRNA expressions of MrgprA3, MrgprC11 and MrgprD in DRG, and up-regulating the protein expressions of Claudin-1 and Occludin in skin tissue.
3.Preparation and application of bovine CD4 monoclonal antibodies.
Wunjun KONG ; Yueshu ZHU ; Zhengzhong XU ; Chengkun ZHENG ; Xiang CHEN ; Xinan JIAO
Chinese Journal of Cellular and Molecular Immunology 2025;41(5):450-455
Objective To prepare monoclonal antibodies against bovine CD4 and identify their basic biological characteristics. Methods Recombinant bovine CD4 (rHis-BoCD4 and rGST-BoCD4) was successfully expressed and purified by constructing a prokaryotic plasmid of bovine CD4 gene. The bovine CD4 monoclonal antibody was produced using hybridoma technology. The subtype and potency of the monoclonal antibody were identified and analyzed by ELISA, while specificity was analyzed through indirect immunofluorescence assay (IFA) and Western-blot. Results Four hybridoma cell lines, namely, 1H4, 6A10, 3F9 and 4G10, stably secreting monoclonal antibodies against BoCD4 were successfully obtained. The subclasses of the monoclonal antibodies subclass 6A10 was IgG2b and the rest of the monoclonal antibodies were of IgM type. Western-blot results showed that the four anti-bovine CD4 mAb strains were able to specifically bind to the bovine CD4 protein expressed in vitro. Indirect immunofluorescence assay showed that four monoclonal antibodies were able to specifically recognize the natural bovine CD4 protein. Flow cytometry assay showed that 3F9 was best to recognize bovine natural CD4 molecules. Conclusion Four monoclonal antibody strains with high specificity to natural bovine CD4 protein were successfully prepared, which lays the foundation for the subsequent studies on the function of bovine CD4 and diagnosis and treatment of bovine T-lymphocyte diseases.
Animals
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Antibodies, Monoclonal/isolation & purification*
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Cattle
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CD4 Antigens/genetics*
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Hybridomas/immunology*
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Antibody Specificity/immunology*
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Mice
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Mice, Inbred BALB C
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Enzyme-Linked Immunosorbent Assay
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Fluorescent Antibody Technique, Indirect
4.Dissecting Social Working Memory: Neural and Behavioral Evidence for Externally and Internally Oriented Components.
Hanxi PAN ; Zefeng CHEN ; Nan XU ; Bolong WANG ; Yuzheng HU ; Hui ZHOU ; Anat PERRY ; Xiang-Zhen KONG ; Mowei SHEN ; Zaifeng GAO
Neuroscience Bulletin 2025;41(11):2049-2062
Social working memory (SWM)-the ability to maintain and manipulate social information in the brain-plays a crucial role in social interactions. However, research on SWM is still in its infancy and is often treated as a unitary construct. In the present study, we propose that SWM can be conceptualized as having two relatively independent components: "externally oriented SWM" (e-SWM) and "internally oriented SWM" (i-SWM). To test this external-internal hypothesis, participants were tasked with memorizing and ranking either facial expressions (e-SWM) or personality traits (i-SWM) associated with images of faces. We then examined the neural correlates of these two SWM components and their functional roles in empathy. The results showed distinct activations as the e-SWM task activated the postcentral and precentral gyri while the i-SWM task activated the precuneus/posterior cingulate cortex and superior frontal gyrus. Distinct multivariate activation patterns were also found within the dorsal medial prefrontal cortex in the two tasks. Moreover, partial least squares analyses combining brain activation and individual differences in empathy showed that e-SWM and i-SWM brain activities were mainly correlated with affective empathy and cognitive empathy, respectively. These findings implicate distinct brain processes as well as functional roles of the two types of SWM, providing support for the internal-external hypothesis of SWM.
Humans
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Memory, Short-Term/physiology*
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Male
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Female
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Empathy/physiology*
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Young Adult
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Magnetic Resonance Imaging
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Adult
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Brain/diagnostic imaging*
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Brain Mapping
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Facial Expression
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Social Behavior
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Facial Recognition/physiology*
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Social Perception
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Personality/physiology*
5.International clinical practice guideline on the use of traditional Chinese medicine for functional dyspepsia (2025).
Sheng-Sheng ZHANG ; Lu-Qing ZHAO ; Xiao-Hua HOU ; Zhao-Xiang BIAN ; Jian-Hua ZHENG ; Hai-He TIAN ; Guan-Hu YANG ; Won-Sook HONG ; Yu-Ying HE ; Li LIU ; Hong SHEN ; Yan-Ping LI ; Sheng XIE ; Jin SHU ; Bin-Fang ZENG ; Jun-Xiang LI ; Zhen LIU ; Zheng-Hua XIAO ; Jing-Dong XIAO ; Pei-Yong ZHENG ; Shao-Gang HUANG ; Sheng-Liang CHEN ; Gui-Jun FEI
Journal of Integrative Medicine 2025;23(5):502-518
Functional dyspepsia (FD), characterized by persistent or recurrent dyspeptic symptoms without identifiable organic, systemic or metabolic causes, is an increasingly recognized global health issue. The objective of this guideline is to equip clinicians and nursing professionals with evidence-based strategies for the management and treatment of adult patients with FD using traditional Chinese medicine (TCM). The Guideline Development Group consulted existing TCM consensus documents on FD and convened a panel of 35 clinicians to generate initial clinical queries. To address these queries, a systematic literature search was conducted across PubMed, EMBASE, the Cochrane Library, China National Knowledge Infrastructure (CNKI), VIP Database, China Biology Medicine (SinoMed) Database, Wanfang Database, Traditional Medicine Research Data Expanded (TMRDE), and the Traditional Chinese Medical Literature Analysis and Retrieval System (TCMLARS). The evidence from the literature was critically appraised using the Grading of Recommendations Assessment, Development, and Evaluation (GRADE) approach. The strength of the recommendations was ascertained through a consensus-building process involving TCM and allopathic medicine experts, methodologists, pharmacologists, nursing specialists, and health economists, leveraging their collective expertise and empirical knowledge. The guideline comprises a total of 43 evidence-informed recommendations that span a range of clinical aspects, including the pathogenesis according to TCM, diagnostic approaches, therapeutic interventions, efficacy assessments, and prognostic considerations. Please cite this article as: Zhang SS, Zhao LQ, Hou XH, Bian ZX, Zheng JH, Tian HH, Yang GH, Hong WS, He YY, Liu L, Shen H, Li YP, Xie S, Shu J, Zeng BF, Li JX, Liu Z, Xiao ZH, Xiao JD, Zheng PY, Huang SG, Chen SL, Fei GJ. International clinical practice guideline on the use of traditional Chinese medicine for functional dyspepsia (2025). J Integr Med. 2025; 23(5):502-518.
Dyspepsia/drug therapy*
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Humans
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Medicine, Chinese Traditional/methods*
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Practice Guidelines as Topic
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Drugs, Chinese Herbal/therapeutic use*
6.Preparation modification strategies for clinical treatment drugs of Parkinson's disease
Meng-jiao HE ; Yi-fang XIAO ; Xiang-an-ni KONG ; Zhi-hao LIU ; Xiao-guang WANG ; Hao FENG ; Jia-sheng TU ; Qian CHEN ; Chun-meng SUN
Acta Pharmaceutica Sinica 2024;59(3):574-580
Parkinson's disease (PD) is a chronic neurodegenerative disease. At present, levodopa and other drugs are mainly used for dopamine supplementation therapy. However, the absorption of levodopa in the gastrointestinal tract is unstable and its half-life is short, and long-term use of levodopa will lead to the end-of-dose deterioration, dyskinesia, the "ON-OFF" phenomenon and other symptoms. Therefore, new preparations need to be developed to improve drug efficacy, reduce side effects or improve compliance of patients. Based on the above clinical needs, this review briefly introduced the preparation modification strategies for the treatment of PD through case analysis, in order to provide references for the research and development of related preparations.
7.A national questionnaire survey on endoscopic treatment for gastroesophageal varices in portal hypertension in China
Xing WANG ; Bing HU ; Yiling LI ; Zhijie FENG ; Yanjing GAO ; Zhining FAN ; Feng JI ; Bingrong LIU ; Jinhai WANG ; Wenhui ZHANG ; Tong DANG ; Hong XU ; Derun KONG ; Lili YUAN ; Liangbi XU ; Shengjuan HU ; Liangzhi WEN ; Ping YAO ; Yunxiao LIANG ; Xiaodong ZHOU ; Huiling XIANG ; Xiaowei LIU ; Xiaoquan HUANG ; Yinglei MIAO ; Xiaoliang ZHU ; De'an TIAN ; Feihu BAI ; Jitao SONG ; Ligang CHEN ; Yingcai MA ; Yifei HUANG ; Bin WU ; Xiaolong QI
Chinese Journal of Digestive Endoscopy 2024;41(1):43-51
Objective:To investigate the current status of endoscopic treatment for gastroesophageal varices in portal hypertension in China, and to provide supporting data and reference for the development of endoscopic treatment.Methods:In this study, initiated by the Liver Health Consortium in China (CHESS), a questionnaire was designed and distributed online to investigate the basic condition of endoscopic treatment for gastroesophageal varices in portal hypertension in 2022 in China. Questions included annual number and indication of endoscopic procedures, adherence to guideline for preventing esophagogastric variceal bleeding (EGVB), management and timing of emergent EGVB, management of gastric and isolated varices, and improvement of endoscopic treatment. Proportions of hospitals concerning therapeutic choices to all participant hospitals were calculated. Guideline adherence between secondary and tertiary hospitals were compared by using Chi-square test.Results:A total of 836 hospitals from 31 provinces (anotomous regions and municipalities) participated in the survey. According to the survey, the control of acute EGVB (49.3%, 412/836) and the prevention of recurrent bleeding (38.3%, 320/836) were major indications of endoscopic treatment. For primary [non-selective β-blocker (NSBB) or endoscopic therapies] and secondary prophylaxis (NSBB and endoscopic therapies) of EGVB, adherence to domestic guideline was 72.5% (606/836) and 39.2% (328/836), respectively. There were significant differences in the adherence between secondary and tertiary hospitals in primary prophylaxis of EGVB [71.0% (495/697) VS 79.9% (111/139), χ2=4.11, P=0.033] and secondary prophylaxis of EGVB [41.6% (290/697) VS 27.3% (38/139), χ2=9.31, P=0.002]. A total of 78.2% (654/836) hospitals preferred endoscopic therapies treating acute EGVB, and endoscopic therapy was more likely to be the first choice for treating acute EGVB in tertiary hospitals (82.6%, 576/697) than secondary hospitals [56.1% (78/139), χ2=46.33, P<0.001]. The optimal timing was usually within 12 hours (48.5%, 317/654) and 12-24 hours (36.9%, 241/654) after the bleeding. Regarding the management of gastroesophageal varices type 2 and isolated gastric varices type 1, most hospitals used cyanoacrylate injection in combination with sclerotherapy [48.2% (403/836) and 29.9% (250/836), respectively], but substantial proportions of hospitals preferred clip-assisted therapies [12.4% (104/836) and 26.4% (221/836), respectively]. Improving the skills of endoscopic doctors (84.2%, 704/836), and enhancing the precision of pre-procedure evaluation and quality of multidisciplinary team (78.9%, 660/836) were considered urgent needs in the development of endoscopic treatment. Conclusion:A variety of endoscopic treatments for gastroesophageal varices in portal hypertension are implemented nationwide. Participant hospitals are active to perform emergent endoscopy for acute EGVB, but are inadequate in following recommendations regarding primary and secondary prophylaxis of EGVB. Moreover, the selection of endoscopic procedures for gastric varices differs greatly among hospitals.
8.Asymmetry of multifidus muscle in patients with unilateral lumbosacral radiculopathy due to lumbar disc herniation and lumbar spondylolisthesis
Chensheng QIU ; Demao KONG ; Yongsheng ZHAO ; Libin FENG ; Hongfei XIANG ; Zhu GUO ; Yuanxue YI ; Bohua CHEN
Chinese Journal of Orthopaedics 2024;44(21):1384-1392
Objective:To investigate the morphological difference and clinical significance of bilateral lumbar multifidus muscles in patients with unilateral lumbosacral radiculopathy due to lumbar disc herniation and lumbar spondylolisthesis.Methods:A retrospective analysis was conducted on patients with low back pain, lumbar disc herniation and lumbar spondylolisthesis. Patients with lumbar disc herniation or lumbar spondylolisthesis underwent single segment lesion either at L 4, 5 or L 5S 1, while those accompanied with unilateral lumbosacral radiculopathy underwent percutaneous endoscopic lumbar discectomy or conventional open surgery at Qingdao Municipal Hospital between January 2017 and January 2023. Patients with lumbar spondylolisthesis were subdivided into degenerative lumbar spondylolisthesis and isthmic spondylolisthesis. 53 patients with low back pain met the inclusion criteria. 170 patients with lumbar disc herniation met the inclusion criteria, with 101 at L 4, 5 and 69 at L 5S 1 level. 129 patients with lumbar spondylolisthesis met the inclusion criteria, including 91 of degenerative lumbar spondylolisthesis at L 4, 5 level and 9 at L 5S 1 level, and 11 of isthmic spondylolisthesis at L 4, 5 level and 18 at L 5S 1 level. Cross-sectional images at the mid-disc of L 3, 4, L 4, 5 and L 5S 1 segments in MRI were acquired. Relative total cross-sectional area (rTCSA), relative functional cross-sectional area (rFCSA), fat infiltration rate (FIR), relative fat distance (rFD) and differential value FIR (D-FIR) in bilateral lumbar multifidus muscle were measured respectively by using Image J software, and were then used to evaluate the atrophy and fat infiltration of bilateral lumbar multifidus muscles. Results:No significant difference was found between the both sides of multifidus muscle in low back pain patients. L 4, 5 lumbar disc herniation group had smaller rFCSA (0.34±0.10 and 0.35±0.10) and larger FIR [29.92(22.21, 36.46) and 26.48(17.54, 34.55)] and rFD [0.39(0.29, 0.54) and 0.32(0.21, 0.43)] on the affected side compared to the unaffected side in L 4, 5 segment, and had larger FIR (34.83±11.34 and 31.44±10.94) and rFD [0.59(0.43, 0.77) and 0.51(0.37, 0.69)] on the affected side in L 5S 1 segment. L 5S 1 lumbar disc herniation group had smaller rFCSA (0.41±0.11 and 0.42±0.12) and larger FIR [26.84(22.92, 35.29) and 24.02(20.03, 32.87)] and rFD (0.51±0.28 and 0.42±0.26) on the affected side in L 5S 1 segment. L 4, 5 degenerative lumbar spondylolisthesis group had larger FIR (36.49±9.76 and 34.72±9.86) on the affected side in L 4, 5 segment, and had larger FIR [35.03(28.64, 41.85) and 33.34(26.37, 39.76)] on the affected side in L 5S 1 segment. L 5S 1 degenerative lumbar spondylolisthesis group had larger FIR [42.53(37.94, 46.81) and 40.79(30.84, 43.53)] and rFD (1.12±0.79 and 0.94±0.79) on the affected side in L 5S 1 segment. L 4, 5 isthmic spondylolisthesis group had smaller rFCSA [0.24(0.20, 0.30) and 0.29(0.23, 0.34)]and larger FIR [34.19 31.30, 42.39) and 29.43(28.82, 36.89)] and rFD (0.39±0.15 and 0.29±0.15) on the affected side in L 4, 5 segment, and had larger FIR (43.18±12.71 and 34.12±11.63) on the affected side in L 5S 1 segment. L 5S 1 isthmic spondylolisthesis group had larger FIR (40.24±9.34 and 36.37±10.70) on the affected side in L 5S 1 segment. No significant difference was found of the multifidus muscle between the affected and unaffected sides in the proximal adjacent segment of the responsible segment in lumbar disc herniation or lumbar spondylolisthesis group patients. L 4, 5 isthmic spondylolisthesis group had larger D-FIR (6.75±8.46 and 1.78±5.77) in L 4, 5 segment, and had larger D-FIR (9.06±11.59 and 1.54±7.08) in L 5S 1 segment compared to L 4, 5 degenerative lumbar spondylolisthesis group. Grade Ⅱ L 4, 5 lumbar spondylolisthesis group had larger D-FIR (10.73±13.61 and 1.92±7.43) in L 5S 1 segment compared to grade Ⅰ L 4, 5 lumbar spondylolisthesis group. Conclusion:L 4, 5 or L 5S 1 lumbar disc herniation and lumbar spondylolisthesis patients with unilateral lumbosacral radiculopathy had asymmetric atrophy and fat infiltration of multifidus muscle. The atrophy and fat infiltration on the affected side showed greater. The asymmetry appeared in the responsible segment and its distal adjacent lumbar segment. Lumbar spondylolisthesis patients with a lager degree of slip or with isthmic type could be accompanied by more severe asymmetry of multifidus muscle.
9.Research progress on the evaluation and intervention of social interaction behaviors in animal models of autism
Minghui KONG ; Liming LU ; Leiying XIANG ; Xiaoyi CHEN ; Zhiru ZHU
Chinese Journal of Comparative Medicine 2024;34(10):169-178
Autism spectrum disorder(ASD)is a highly heterogeneous neurodevelopmental disorder with a complex underlying genetic structure.Current preclinical trials,however,mainly rely on rodent models to test the effects of non-pharmacological and pharmacological interventions on the core and related symptoms of ASD.This paper considers the brain regions that affect social interaction behaviors from the perspective of cognitive neural mechanisms,and reviews behavioral testing experiments,such as the three-chamber social interaction test,visible burrow system,and eco-HAB system.We also summarize effective non-pharmacological and pharmacological interventions,such as baclofen,oxytocin,and metformin,in the core and related symptom areas of ASD.This review aims to provide reference directions to promote the development of preclinical trials using rodent models.
10.Mechanism of Colquhounia Root Tablets in inhibiting osteoclast differentiation based on HSP90 target modulation.
Pei-Ping CHEN ; Qian WANG ; Feng-Yu HUANG ; Xiang-Ying KONG ; Na LIN ; Xiao-Hui SU
China Journal of Chinese Materia Medica 2024;49(23):6389-6398
This study aimed to investigate the potential role of Colquhounia Root Tablets against bone destruction in rheumatoid arthritis(RA) and its molecular mechanism. The study used ultra-performance liquid chromatography-mass spectrometry to analyze the major components of Colquhounia Root Tablets and predicted its candidate target gene set based on the major components. The key targets of RA bone destruction were obtained through GeneCards and the Database of Genetics and Medical Literature(OMIM), protein-protein interaction(PPI) network was constructed, and the key targets were identified by topological analysis. The molecular mechanism of Colquhounia Root Tablets against RA bone destruction was further revealed using Gene Ontology(GO) and Kyoto Encyclopedia of Genes and Genomes(KEGG) enrichment analysis. The effects of Colquhounia Root Tablets on macrophage viability was assessed by MTS assay and screened for non-toxic concentrations. A model of receptor activator of nuclear factor-κB(RANKL) induced osteoclast differentiation in vitro was constructed. Colquhounia Root Tablets were used to observe the formation and differentiation of osteoclasts by tartrate-resistant acid phosphatase(TRAP) staining and fibrous actin(F-actin) staining, and the effects of Colquhounia Root Tablets on the changes of core target proteins in the osteoclast differentiation system were detected by immunofluorescence and Western blot. The results showed that the main components of Colquhounia Root Tablets included 14 compounds such as triptolide, celastrol, and triptophenolide. Further network analysis revealed that heat-shock protein 90(HSP90) was the key target gene of Colquhounia Root Tablets for anti-RA bone destruction. TRAP staining and F-actin staining showed that the number and area of TRAP-positive polymorphonuclear cells, as well as actin rings, were reduced in a dose-dependent manner after the intervention of Colquhounia Root Tablets(P<0.01). Western blot results showed that the expression of HSP90 protein was significantly reduced after intervention with Colquhounia Root Tablets at 20 and 40 μg·mL~(-1)(P<0.01); Colquhounia Root Tablets at 10 μg·mL~(-1) could significantly decrease the expression of necrosis factor receptor associated molecule 6(TRAF6) and nuclear factor of activated T cells 1(NFATc1) proteins(P<0.01); moreover, all doses of Colquhounia Root Tablets significantly reduced the expression of osteoclast differentiation marker proteins matrix metalloproteinase 9(MMP9) and cathepsin K(CTSK)(P<0.01).Immunofluorescence results further confirmed that Colquhounia Root Tablets significantly inhibited HSP90 and CTSK levels, as well as NFATc1 activation in osteoblasts. In conclusion, the present study confirmed that Colquhounia Root Tablets may inhibit RANKL-induced osteoclast differentiation by regulating the key target of HSP90, thus exerting an anti-RA bone destruction effect, which will provide a new idea for Colquhounia Root Tablets to prevent and treat bone destruction in rheumatoid arthritis.
Osteoclasts/metabolism*
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Mice
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Animals
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Cell Differentiation/drug effects*
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HSP90 Heat-Shock Proteins/genetics*
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Drugs, Chinese Herbal/chemistry*
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Plant Roots/chemistry*
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Humans
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Arthritis, Rheumatoid/physiopathology*
;
Protein Interaction Maps/drug effects*

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