摘要：
Objective:To observe the outcome of Gugutou Huaisiyu Jiaonang(股骨头坏死愈胶囊,GGTHSYJN)in treatment of ster-oid-induced osteonecrosis of femoral head(SONFH),and to explore its underlying mechanism via multiplex immunohistochemistry(mIHC)technique.Methods:One hundred Sprague-Dawley(SD)rats were randomly assigned into normal control group,model group,low-dose drug treatment group and high-dose drug treatment group.After adaptive feeding for one week,the rats in model group,low-dose drug treatment group and high-dose drug treatment group were subjected to intramuscular injection of lipopolysaccharide and methylprednisolone in turn for inducing SONFH.After the end of modeling,the rats in low-dose drug treatment group were intragastric administrated with 0.33 g/kg GGTHSYJN solution(GGTHSYJN powders were dissolved into water),the ones in high-dose drug treatment group with 0.67 g/kg GGTH-SYJN solution,while the ones in normal control group and model group with the same dose of normal saline.All rats in the 4 groups were in-tragastric administrated twice a day for consecutive 4 weeks.After the end of treatment,the femur heads were harvested from the rats,and the cancellous bone microstructure of femur head was observed and analyzed by using Micro-CT scanning;meanwhile,the tissue sections of femur head were stained with hematoxylin-eosin(HE)for observing the histopathological changes.Furthermore,the macrophage polarization and the expressions of Toll-like receptor 4(TLR4)/myeloid differentiation primary response gene 88(MyD88)/nuclear factor-KB(NF-κB)signaling pathway-related protein were analyzed by using mIHC technique.Results:①One rat in model group died after the modeling,and 3 rats in low-dose drug treatment group and 2 ones in high-dose drug treatment group died during the treatment.Rats with good mental state,normal eating and drinking as well as healthy fur without shedding were observed in normal group.On day 3 after the beginning of the modeling,the rats in model group,low-dose drug treatment group,and high-dose drug treatment group gradually exhibited the symptoms as dispiritedness,reduced eating and drinking,as well as slight hair removal,and the signs gradually recovered after one week.②The normal control group exhibited higher bone volume fraction(BVF),thicker trabeculae,more trabeculae,and lower trabecular separation(Tb.Sp)in cancellous bone compared with that of model group(P=0.000,P=0.000,P=0.000,P=0.000).The differences in BVF and trabecular thickness were not statistically significant between low-dose drug treatment group and model group(P=0.052,P=0.071),while,the trabe-culae was more and the Tb.Sp was lower in low-dose drug treatment group compared to model group(P=0.012;P=0.001).The high-dose drug treatment group displayed higher BVF,thicker trabeculae,more trabeculae,and lower Tb.Sp in cancellous bone compared with that of model group(P=0.001,P=0.011,P=0.000,P=0.001),while,in contrast to that of low-dose drug treatment group,the results revealed no significant differences(P=0.146,P=0.414,P=0.086,P=0.146).③The percentage of empty lacunae and the incidence rate of os-teonecrosis in femur heads were higher in model group compared to normal control group(P=0.000,P=0.000).The percentage of empty lacunae in femur head was lower in low-dose drug treatment group compared to model group(P=0.000),while,the comparison of osteone-crosis incidence rate between the 2 groups revealed no significant difference(P=0.054).The percentage of empty lacunae and the inci-dence rate of osteonecrosis in femur heads were lower in high-dose drug treatment group compared to model group(P=0.000,P=0.000);furthermore,the percentage of empty lacunae was lower in high-dose drug treatment group compared to low-dose drug treatment group(P=0.049),while,the comparison of osteonecrosis incidence rate between the 2 groups revealed no significant difference(P=0.556).④The M1 macrophages and M2 macrophages accounted for a higher proportion in femur heads of rats in model group compared to normal control group(P=0.000,P=0.000).There was no statistical difference in the proportions of M1 macrophages and M2 macrophages between low-dose drug treatment group and model group(P=0.270,P=0.533).The M1 macrophages accounted for a lower proportion in high-dose drug treatment group compared to model group(P=0.009),while the M2 macrophages accounted for a higher proportion in high-dose drug treatment group compared to model group and low-dose drug treatment group(P=0.006,P=0.038);furthermore,the comparison of the proportion of M1 macrophages between high-dose drug treatment group and low-dose drug treatment group revealed no significant difference(P=0.131).⑤The relative expression levels of TLR4,MyD88 and NF-κB p65 protein in femur heads were higher in model group com-pared to normal control group(P=0.000,P=0.000,P=0.000).There was no statistical difference in the relative expression levels of TLR4 and MyD88 protein between low-dose drug treatment group and model group(P=0.268,P=0.280),while the relative expression level of NF-κB p65 protein was lower in low-dose drug treatment group compared to model group(P=0.034).The relative expression levels of TLR4,MyD88 and NF-κB p65 protein were lower in high-dose drug treatment group compared to model group and low-dose drug treat-ment group(TLR4:P=0.002,P=0.040;MyD88:P=0.000,P=0.013;NF-κB p65:P=0.000,P=0.039).Conclusion:GGTHSYJN can significantly improve the bone microstructure of femur head and inhibit osteonecrosis in treatment of SONFH,and it exhibits dose-dependence in the efficacy.It may work by regulating TLR4/MyD88/NF-KB signaling pathway and macrophage polarization.
摘要：
探讨柴金解郁安神片调控前扣带皮层（ACC）-腹侧海马（vHPC）谷氨酸能神经环路异常改善抑郁症大鼠腹侧海马神经元突触重塑的分子机制。首先运用化学遗传将谷氨酸能腺相关病毒（AAV）定位注射至大鼠ACC脑区，并通过慢性温和不可预知性应激（CUMS）联合孤笼饲养复制大鼠抑郁模型，实验设正常组、模型组、AAV空载组、AAV病毒组、AAV病毒+糖皮质激素受体（GR）阻断剂组、AAV病毒+趋化因子受体1（CX3CR1）阻断剂组、AAV病毒+柴金解郁安神片组，采用水迷宫（Morris water maze）、旷场（open-field）和强迫游泳（forced-swimming）试验联合动物行为分析系统评估大鼠抑郁样行为；苏木素-伊红（HE）染色检测大鼠ACC及vHPC脑区神经元形态结构变化；免疫荧光及核磷酸蛋白（c-Fos）检测大鼠ACC-vHPC谷氨酸能神经环路激活情况；高尔基染色和透射电镜检测大鼠vHPC神经元树突、树突棘及突触亚微结构变化；免疫荧光、Western blot分别检测大鼠vHPC谷氨酸能神经元细胞内突触重塑相关蛋白谷氨酸受体2A（GRIN2A）、谷氨酸受体2B（GRIN2B）、Ca2+/钙调蛋白依赖性蛋白激酶Ⅱ（CaMKⅡ）、丝裂原激活蛋白激酶激活蛋白激酶2（MK2）、丝切蛋白（cofilin）表达水平。结果表明，谷氨酸能AAV病毒激活后模型组大鼠抑郁样行为表型、ACC及vHPC神经元形态结构、突触超微结构损伤更加加重，而GR、CX3CR1阻断剂均能不同程度逆转其异常改变，提示ACC脑区内胶质细胞GR/CX3CR1双信号介导的ACC-vHPC谷氨酸能神经环路异常激活可能与抑郁的发生发展密切相关。有趣的是，柴金解郁安神片也能显著抑制AAV病毒诱导的ACC-vHPC神经环路激活及Glu含量异常升高，同时有效逆转模型组大鼠进一步加重的抑郁样行为和vHPC谷氨酸能神经元突触重塑，并揭示其改善腹侧海马神经元突触损伤的分子机制可能与调控突触重塑相关信号NR/CaMKⅡ、MK2/Cofilin有关。综上，本文证实了柴金解郁安神片能有效调控ACC-vHPC谷氨酸能神经环路异常进而改善抑郁症大鼠腹侧海马谷氨酸能神经元突触重塑，其分子机制可能与调节突触相关NR/CaMKⅡ、MK2/Cofilin信号通路有关，这可能是其发挥抗抑郁作用的重要机制。
期刊：
Frontiers in Microbiology,2024年15:1354823 ISSN：1664-302X
通讯作者：
Tan, ZJ;He, WZ
作者机构：
[Guo, Mingmin] Hunan Univ Chinese Med, Sch Pharm, Changsha, Peoples R China.;[Shen, Junxi; Tan, Zhoujin; Fang, Leyao] Hunan Univ Chinese Med, Sch Tradit Chinese Med, Changsha, Peoples R China.;[Chen, Meili] Changsha Hosp Tradit Chinese Med, Changsha, Peoples R China.;[He, Wenzhi] Hunan Univ Chinese Med, Sch Stomatol, Changsha, Peoples R China.
通讯机构：
[He, WZ ; Tan, ZJ ] H;Hunan Univ Chinese Med, Sch Tradit Chinese Med, Changsha, Peoples R China.;Hunan Univ Chinese Med, Sch Stomatol, Changsha, Peoples R China.
关键词：
CutC;TMAO;adenine;cecal microbiota;diarrhea with kidney-yang deficiency syndrome;folium sennae;inflammatory factors
摘要：
OBJECTIVE: Previous studies have indicated that diarrhea with kidney-yang deficiency syndrome leads to a disorder of small intestine contents and mucosal microbiota. However, the relationship of TMA-lyase (CutC) activity and TMAO with diarrhea with kidney-yang deficiency syndrome remains unexplored. Therefore, this study explores the relationship between cecal microbiota and choline TMA-lyase (CutC) activity, as well as the correlation between trimethylamine oxide (TMAO), inflammatory index, and CutC activity. METHOD: Twenty SPF-grade male KM mice were randomly divided into the normal group (CN) and the diarrhea model group (CD). Diarrhea mouse models were established by adenine combined with Folium sennae administration. CutC activity, TMAO, interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α) levels were detected, and the cecal content microbiota was sequenced. RESULT: After 14 days, diarrhea occurred in the CD group. Compared with the CN group, there was no significant change in the activity of CutC in the small intestine of the CD group, while the activity of CutC in the cecum was significantly increased, and the levels of TMAO, IL-6, and TNF-α showed a significant increase. The Chao1 index, Observed_species index, Shannon index, and Simpson index all exhibited a decreasing trend. The main changes at the bacterial genus level were Alistipes, Enterorhabdus, Desulfovibrio, Bacteroides, Candidatus_Saccharimonas, and [Ruminococcus]_torques_group. The results of LEfSe analysis, random forest analysis and ROC curve analysis revealed Paludicola, Blautia, Negativibacillus, Paraprevotella, Harryflintia, Candidatus_Soleaferrea, Anaerotruncus, Oscillibacter, Colidextribacter, [Ruminococcus]_torques_group, and Bacteroides as characteristic bacteria in the CD group. Correlation analysis showed a significant negative correlation between cecal CutC activity and Ligilactobacillus, and a significant positive correlation with Negativibacillus and Paludicola. The level of TMAO was significantly positively correlated with CutC activity and IL-6. CONCLUSION: Diarrhea with kidney-yang deficiency syndrome significantly affects the physiological status, digestive enzyme activity, CutC activity, TMAO levels, and inflammatory response in mice. Additionally, there are changes in the composition and function of cecal microbiota, indicating an important impact of diarrhea with kidney-yang deficiency syndrome on the host intestinal microbiota balance. The occurrence of diarrhea with kidney-yang deficiency syndrome may be associated with dysbiosis of intestinal microbiota, increased CutC activity, elevated TMAO levels, and heightened inflammatory factor levels.
期刊：
Journal of Ginseng Research,2024年 ISSN：1226-8453
通讯作者：
Hongxin Wang
作者机构：
[Chenyang Ran; Meili Lu; Yi Hao; Xinyu Guo; Yunhan Li; Hongxin Wang] The Key Laboratory of Cardiovascular and Cerebrovascular Drug Research of Liaoning Province, Jinzhou Medical University, Jinzhou, 121001, PR China;[Fang Zhao] Institute of Innovation and Applied Research in Chinese Medicine and Department of Rheumatology of the First Hospital, Hunan University of Chinese Medicine. Changsha, Hunan, 410208, PR China;[Yuhong Su] The College of Food and Health of Liaoning Province, Jinzhou Medical University, Jinzhou, PR China
通讯机构：
[Hongxin Wang] T;The Key Laboratory of Cardiovascular and Cerebrovascular Drug Research of Liaoning Province, Jinzhou Medical University, Jinzhou, 121001, PR China
摘要：
Hypoxic pulmonary hypertension (HPH) is the main pathological change in vascular remodeling, a complex cardiopulmonary disease caused by hypoxia. Some research results have shown that ginsenoside Rg1 (Rg1) can improve vascular remodeling, but the effect and mechanism of Rg1 on hypoxia-induced pulmonary hypertension are not clear. The purpose of this study was to discuss the potential mechanism of action of Rg1 on HPH.
C57BL/6 mice, calpain-1 knockout mice and Pulmonary artery smooth muscle cells (PASMCs) were exposed to a low oxygen environment with or without different treatments. The effect of Rg1 and calpain-1 silencing on inflammation, fibrosis, proliferation and the protein expression levels of calpain-1, STAT3 and p-STAT3 were determined at the animal and cellular levels.
At the mouse and cellular levels, hypoxia promotes inflammation, fibrosis, and cell proliferation, and the expression of calpain-1 and p-STAT3 is also increased. Ginsenoside Rg1 administration and calpain-1 knockdown, MDL-28170, and HY-13818 treatment showed protective effects on hypoxia-induced inflammation, fibrosis, and cell proliferation, which may be associated with the downregulation of calpain-1 and p-STAT3 expression in mice and cells. In addition, overexpression of calpain 1 increased p-STAT3 expression, accelerating the onset of inflammation, fibrosis and cell proliferation in hypoxic PASMCs.
Ginsenoside Rg1 may ameliorate hypoxia-induced pulmonary vascular remodeling by suppressing the calpain-1/STAT3 signaling pathway.
期刊：
BMC Complementary Medicine and Therapies,2024年24(1):1-12 ISSN：2662-7671
通讯作者：
Liqing Li<&wdkj&>Bin Liu<&wdkj&>Xiong Cai
作者机构：
[Zhaoli Su; Junping Zhu; Ye Lin; Yuanyuan Tang; Jiaming Wei] Department of Rheumatology, First Hospital, School of Chinese Medical Sciences, Hunan University of Chinese Medicine, Changsha, Hunan, 410208, China;[Yuanyuan Tang] College of Biology, Hunan University, Changsha, Hunan, 410082, China;[Zhaoli Su] The Central Research Laboratory, Hunan Traditional Chinese Medical College, Zhuzhou, China;[Zhaoli Su] Guangxi Provincial Key Laboratory of Preventive and Therapeutic Research in Prevalent Diseases in West Guangxi, Youjiang Medical University for Nationalities, Baise, Guangxi, 533000, China;[Liqing Li] The Central Research Laboratory, Hunan Traditional Chinese Medical College, Zhuzhou, China. liliqing87@qq.com
通讯机构：
[Liqing Li] T;[Bin Liu] C;[Xiong Cai] D;The Central Research Laboratory, Hunan Traditional Chinese Medical College, Zhuzhou, China<&wdkj&>Guangxi Provincial Key Laboratory of Preventive and Therapeutic Research in Prevalent Diseases in West Guangxi, Youjiang Medical University for Nationalities, Baise, China<&wdkj&>College of Biology, Hunan University, Changsha, China<&wdkj&>Department of Rheumatology, First Hospital, School of Chinese Medical Sciences, Hunan University of Chinese Medicine, Changsha, China
摘要：
BACKGROUND: Rheumatoid arthritis (RA) is a prevalent autoimmune disease marked by chronic synovitis as well as cartilage and bone destruction. Halofuginone hydrobromide (HF), a bioactive compound derived from the Chinese herbal plant Dichroa febrifuga Lour., has demonstrated substantial anti-arthritic effects in RA. Nevertheless, the molecular mechanisms responsible for the anti-RA effects of HF remain unclear. METHODS: This study employed a combination of network pharmacology, molecular docking, and experimental validation to investigate potential targets of HF in RA. RESULTS: Network pharmacology analyses identified 109 differentially expressed genes (DEGs) resulting from HF treatment in RA. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses unveiled a robust association between these DEGs and the IL-17 signaling pathway. Subsequently, a protein-protein interaction (PPI) network analysis revealed 10 core DEGs, that is, EGFR, MMP9, TLR4, ESR1, MMP2, PPARG, MAPK1, JAK2, STAT1, and MAPK8. Among them, MMP9 displayed the greatest binding energy for HF. In an in vitro assay, HF significantly inhibited the activity of inflammatory macrophages, and regulated the IL-17 signaling pathway by decreasing the levels of IL-17C, p-NF-κB, and MMP9. CONCLUSION: In summary, these findings suggest that HF has the potential to inhibit the activation of inflammatory macrophages through its regulation of the IL-17 signaling pathway, underscoring its potential in the suppression of immune-mediated inflammation in RA.
