1.Potential Mechanism of Zuojinwan in Improving Liver Fibrosis Based on Hepatic Tissue Metabolomics
Yiting JIANG ; Kexin LIU ; Yixi QIAN ; Rui ZHANG ; Feng ZHANG ; Hongyan WU ; Li CHEN
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(6):54-61
ObjectiveThis study aims to elucidate the potential mechanism of Zuojinwan in improving liver fibrosis through hepatic tissue metabolomics analysis. MethodsTwenty-four mice were randomly allocated into normal group, model group , positive drug group (silymarin, 100 mg·kg-1), and Zuojinwan group (Zuojinwan solution, 2.5 g·kg-1), with per group six mice. Liver fibrosis model was induced via intraperitoneal injection of olive oil solution with 10% carbon tetrachloride (CCl4) (0.5 μL·g-1, three times weekly for 8 weeks) in all groups except the normal group. During the final 4 weeks, the silymarin group received silymarin (100 mg·kg-1) by gavage thrice weekly, while the Zuojinwan group was administered Zuojinwan solution (2.5 g·kg-1) under the same regimen. After the last administration, the levels of liver fibrosis indicators and liver injury markers in serum were detected. The pathological morphological changes of the liver tissues were observed. The levels of liver fibrosis markers α-smooth muscle actin (α-SMA) and Collagen Ⅰ(ColⅠ) were detected. Metabolomics was analyzed on mice's liver tissues. The mice's serum was collected for metabolomics analysis. ResultsCompared with the model group, Zuojinwan significantly improved indicators related to liver fibrosis and liver injury. Compared with the normal group, the model group showed significantly elevated levels of fibrosis markers such as laminin (LN), hyaluronic acid (HA), procollagen typeⅢ (PC-Ⅲ), and type Ⅳ Collagen (Ⅳ-C), while liver injury indicators such as alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), lactate dehydrogenase (LDH), and total bilirubin (TBIL), exhibited a marked upward trend (P<0.05). Compared with the model group, the silymarin group showed a significant decrease in the aforementioned indicators (P<0.05). Notably, compared with the model group, the Zuojinwan group exhibited a significant reduction in all these indicators (P<0.05), with efficacy comparable to that of the silymarin group. Zuojinwan reduced mRNA and protein levels of α-SMA and ColⅠ in the liver tissue. Metabolomics results revealed that compared with the model group, Zuojiinwan significantly reduced levels of glucose metabolism-related metabolites such as D-fructose 1,6-bisphosphate (FBP), nicotinamide adenine dinucleotide phosphate (NADPH), sodium beta-D-fructose 6-phosphate (F6P), dihydroxyacetone phosphate (DHAP), fumaric acid, and D-glucose 6-phosphate (G6P) (P<0.05). Serum enzyme-linked immunosorbent assay (ELISA) was used to detect glucose metabolism indicators and further validate the regulatory effect of Zuojinwan on glucose metabolism. ConclusionThese results suggest that Zuojinwan may improve liver fibrosis by regulating the dysregulated levels of glucose metabolism during the progression of liver fibrosis.
2.Impact of anticentromere antibody on the clinical features and prognosis of patients with primary biliary cholangitis
Shengzhu HE ; Guiqin ZHOU ; Kexin QIAO ; Yaxing LIU ; Bin LI ; Ying FENG ; Xianbo WANG
Journal of Clinical Hepatology 2025;41(5):872-877
ObjectiveTo investigate the impact of anticentromere antibody (ACA) on the clinical features and prognosis of patients with primary biliary cholangitis (PBC) by comparing clinical classification, ursodeoxycholic acid (UDCA) response, GLOBE score, and UK-PBC score between ACA-positive PBC patients and ACA-negative PBC patients. MethodsA total of 749 patients who were admitted to Beijing Ditan Hospital, Capital Medical University, from August 2013 to December 2022 and were diagnosed with PBC were enrolled and divided into ACA-positive group with 147 patients and ACA-negative group with 602 patients. According to their conditions on admission, the two groups were compared in terms of the distribution of clinical types, i.e., chronic progression-type PBC, portal hypertension-type PBC, and standard jaundice/liver failure-type PBC. There were 261 patients with complete data after 1-year follow-up, among whom there were 53 patients with positive ACA and 208 with negative ACA. A statistical analysis was performed, and propensity score matching was performed based on sex and age at a ratio of 1∶2. The two groups were compared in terms of 1-year UDCA response rate, GLOBE score, and UK-PBC score before and after matching. The independent-samples t test or the Mann-Whitney U test was used for comparison of continuous data between two groups, and the chi-square test was used for comparison of categorical data between two groups. ResultsCompared with the ACA-negative group, the ACA-positive group had a significantly higher age (61.28±10.35 years vs 56.74±12.17 years, t=4.164, P<0.001), a significantly higher proportion of female patients (93.9% vs 77.6%, χ2=20.221, P<0.001), a significantly higher proportion of patients with portal hypertension (48.3% vs 27.6%, χ2=23.289, P<0.001), and a significantly lower proportion of patients with jaundice/liver failure (24.5% vs 38.5%, χ2=10.205, P<0.001). After 1-year follow-up, for the 261 PBC patients with complete data, there was no significant difference in UDCA response rate before propensity score matching between the ACA-positive group and the ACA-negative group (41.5% vs 41.8%, P>0.05), and there was a significant difference in the proportion of patients with a GLOBE score of >0.3 between the ACA-positive group and the ACA-negative group (92.5% vs 80.3%, χ2=3.935, P=0.047). There were 53 patients in the ACA-positive group and 106 patients in the ACA-negative group after propensity score matching, and there were no significant differences between the two groups in UDCA response rate, GLOBE score, and UK-PBC score (all P>0.05). ConclusionACA-positive patients tend to have an older age, with a higher proportion of female patients or patients with portal hypertension, while there is a relatively low proportion of patients with jaundice/liver failure. Positive ACA has no significant impact on UDCA response rate, GLOBE score, and UK-PBC score.
3.Guidelines for Medical Examination for Cancer in Health Examination Agency(2025 Edition)
Wanqing CHEN ; Zhijian XU ; Qiang ZENG ; Ni LI ; Wei CAO ; Kexin CHEN ; Feng SUN ; Yuping LIU ; Yutong HE ; Peng WANG ; Shiqi TANG ; Qun ZHANG ; Kaifeng PAN ; Jie HE
China Cancer 2025;34(9):667-697
Cancer incidence in China has been rising steadily,with a particularly heavy burden from several high-prevalence malignancies.Medical examination for cancer plays a critical role in the early detection of cancer,precancerous lesions,and precursor conditions,thereby facilitating timely diagnosis and intervention.Such examination also addresses the growing demand for person-alized cancer screening services among diverse population groups.The development of evidence-based,context-specific cancer screening guidelines is essential to enhance the standardization,quality,and equity of preventive screening practices across the country,ultimately improving out-comes in early cancer detection and treatment.Guided by the Department of Medical Emergency Response of the National Health Commission,the Guidelines for Medical Examination for Cancer in Health Examination Agency(2025 Edition)were developed under the leadership of the National Cancer Center.A multidisciplinary panel of experts formulated the guidelines in accordance with the principles and methodology of the World Health Organization Handbook for Guideline Deve-lopment.The guidelines provide evidence-based recommendations on key clinical domains:target cancers and populations,overall screening workflow,screening protocols,diagnostic technolo-gies,result interpretation,follow-up procedures,and quality control.The primary objective is to standardize cancer screening practices in health examination agency and strengthen China's ca-pacity for prevention and control of high-burden cancers.
4.Clinical effects of various special forms of the descending branch of the lateral circumflex femoral artery perforator flaps in repairing high-voltage electrical burn wounds on the wrist
Weili DU ; Feng XIONG ; Kexin CHE ; Lin CHENG ; Qiang DAI ; Yuming SHEN
Chinese Journal of Burns 2025;41(1):18-27
Objective:To explore the clinical effects of various special forms of the descending branch of the lateral circumflex femoral artery (DLCFA) perforator flaps in repairing high-voltage electrical burn wounds on the wrist.Methods:This study was a retrospective observational study. From September 2014 to June 2024, 79 male patients with high-voltage electrical burns on the wrist, aged 20 to 62 years and met the inclusion criteria, were admitted to Beijing Jishuitan Hospital Affiliated to Capital Medical University, with wrist high-voltage electrical burn wound (hereinafter referred to as wrist wound) types being classified as type Ⅱ or type Ⅲ. In the early stage after injury, debridement was performed on the patients' wrists. Based on the wound condition and flap indications, the flow-through, lobed, chimeric, flow-through-lobed, lobed-chimeric, flow-through-chimeric, or flow-through-lobed-chimeric DLCFA perforator flap was employed individually, and the flow-through-chimeric DLCFA perforator flap and tensor fascia lata myocutaneous flap were employed in combination to repair the wounds. The donor site wounds were repaired using direct sutures or skin grafting. The number of various DLCFA perforator flaps resected during surgery and the number of various types of wrist wounds repaired were recorded, as well as the closure status of the donor site wound. The postoperative flap survival, occurrence of vascular crisis, wound or suture site healing, and patency of the reconstructed artery in flow-through flaps were recorded. During follow-up, the appearance of the flap, scar formation, and the presence of thigh muscle herniation were observed.Results:Intraoperatively, 11 flow-through DLCFA perforator flaps were resected to repair 11 type Ⅱ wrist wounds, 13 lobed DLCFA perforator flaps were resected to repair 9 type Ⅱ and 4 type Ⅲ wrist wounds, 16 chimeric DLCFA perforator flaps were resected to repair 16 type Ⅱ wrist wounds, 11 flow-through-lobed DLCFA perforator flaps were resected to repair 5 type Ⅱ and 6 type Ⅲ wrist wounds, 10 lobed-chimeric DLCFA perforator flaps were resected to repair 5 type Ⅱ and 5 type Ⅲ wrist wounds, 6 flow-through-chimeric DLCFA perforator flaps were resected to repair 6 type Ⅱ wrist wounds, 7 flow-through-lobed-chimeric DLCFA perforator flaps were resected to repair 7 type Ⅲ wrist wounds, and 5 flow-through-chimeric DLCFA perforator flaps combined with tensor fascia lata myocutaneous flaps were resected to repair 5 type Ⅲ wrist wounds. Seventy-four patients had their donor site wounds closed by direct suturing, while 5 patients had their donor site wounds closed by skin grafting. Postoperatively, the flaps in 3 patients developed vascular crisis, including 1 case of arterial crisis and 2 cases of venous crises but survived after emergency vascular exploration and other treatments; the remaining flaps survived completely. Postoperatively, 3 patients had seepage beneath their flaps, which were closed after dressing changes; the remaining patients' wounds or suture sites all healed. Anteriography showed that all reconstructed arteries in 35 patients who underwent flow-through flap transplantation were patent postoperatively. During the follow-up period of 3 months to 1 year, 20 patients had bloated flap, while the rest had good flap appearance; linear scars were left in the donor sites that underwent direct wound closure, and the skin-grafted areas of the donor site wounds showed no significant patchy hypertrophic scarring; no thigh muscle herniation occurred.Conclusions:Taking the full advantage of perforator flaps, various special forms of the DLCFA perforator flaps are used to repair the three-dimensionally injury wounds caused by high-voltage electrical burns on the wrist, which not only minimizes the damage to the donor site but also allow the recipient site to be well repaired, showing good appearance in the recent follow-up.
5.Impact of anticentromere antibody on the clinical features and prognosis of patients with primary biliary cholangitis
Shengzhu HE ; Guiqin ZHOU ; Kexin QIAO ; Yaxing LIU ; Bin LI ; Ying FENG ; Xianbo WANG
Journal of Clinical Hepatology 2025;42(5):872-877
Objective To investigate the impact of anticentromere antibody(ACA)on the clinical features and prognosis of patients with primary biliary cholangitis(PBC)by comparing clinical classification,ursodeoxycholic acid(UDCA)response,GLOBE score,and UK-PBC score between ACA-positive PBC patients and ACA-negative PBC patients.Methods A total of 749 patients who were admitted to Beijing Ditan Hospital,Capital Medical University,from August 2013 to December 2022 and were diagnosed with PBC were enrolled and divided into ACA-positive group with 147 patients and ACA-negative group with 602 patients.According to their conditions on admission,the two groups were compared in terms of the distribution of clinical types,i.e.,chronic progression-type PBC,portal hypertension-type PBC,and standard jaundice/liver failure-type PBC.There were 261 patients with complete data after 1-year follow-up,among whom there were 53 patients with positive ACA and 208 with negative ACA.A statistical analysis was performed,and propensity score matching was performed based on sex and age at a ratio of 1∶2.The two groups were compared in terms of 1-year UDCA response rate,GLOBE score,and UK-PBC score before and after matching.The independent-samples t test or the Mann-Whitney U test was used for comparison of continuous data between two groups,and the chi-square test was used for comparison of categorical data between two groups.Results Compared with the ACA-negative group,the ACA-positive group had a significantly higher age(61.28±10.35 years vs 56.74±12.17 years,t=4.164,P<0.001),a significantly higher proportion of female patients(93.9%vs 77.6%,χ2=20.221,P<0.001),a significantly higher proportion of patients with portal hypertension(48.3%vs 27.6%,χ2=23.289,P<0.001),and a significantly lower proportion of patients with jaundice/liver failure(24.5%vs 38.5%,χ2=10.205,P<0.001).After 1-year follow-up,for the 261 PBC patients with complete data,there was no significant difference in UDCA response rate before propensity score matching between the ACA-positive group and the ACA-negative group(41.5%vs 41.8%,P>0.05),and there was a significant difference in the proportion of patients with a GLOBE score of>0.3 between the ACA-positive group and the ACA-negative group(92.5%vs 80.3%,χ2=3.935,P=0.047).There were 53 patients in the ACA-positive group and 106 patients in the ACA-negative group after propensity score matching,and there were no significant differences between the two groups in UDCA response rate,GLOBE score,and UK-PBC score(all P>0.05).Conclusion ACA-positive patients tend to have an older age,with a higher proportion of female patients or patients with portal hypertension,while there is a relatively low proportion of patients with jaundice/liver failure.Positive ACA has no significant impact on UDCA response rate,GLOBE score,and UK-PBC score.
6.Exploration on the Effects of Bushen Huoxue Prescription on Rabbit Intervertebral Disc Degeneration Based on the RIPK1/RIPK3/MLKL Signaling Pathway
Wei PENG ; Liguo ZHU ; Xunlu YIN ; Jie LUO ; Kexin YANG ; Minshan FENG ; Jie YU ; Long LIANG ; Linghui LI ; Jiawen ZHAN ; Tao HAN ; Mingyi LUO ; Dian ZHANG
Chinese Journal of Information on Traditional Chinese Medicine 2025;32(2):71-77
Objective To observe the effects of Bushen Huoxue Prescription on the pathway related to necroptosis of nucleus pulposus cells in a model rabbit of intervertebral disc degeneration;To explore its mechanisms in delaying intervertebral disc degeneration.Methods A intervertebral disc degeneration rabbit model was established using the spinal instability method.Totally 40 model rabbits were randomly divided into model group,ibuprofen group and Bushen Huoxue Prescription low-,medium-and high-dosage groups.Additionally,a normal control group and a sham-operation group were set up,with 8 rabbits in each group.Each treatment groups received the corresponding drugs via gavage for two consecutive weeks.HE staining was used to observe morphology of nucleus pulposus tissue,transmission electron microscopy was used to observe ultrastructure in nucleus pulposus cells,immunohistochemical staining was used to assess the expressions of Aggrecan and Collagen Ⅱ in nucleus pulposus tissue,Western blot was used to detect the expressions of p-RIPK1,p-RIPK3 and p-MLKL protein in nucleus pulposus tissue.Results Compared with the sham-operation group,the model group showed a significant decrease in nucleus pulposus cells,disordered cell arrangement,reduced extracellular matrix,interrupted cell membrane continuity under transmission electron microscopy,organelle swelling,nuclear membrane disruption,partial chromatin loss,and positive expression of Aggrecan and Collagen Ⅱ in nucleus pulposus tissue decreased(P<0.01),while the expressions of p-RIPK1,p-RIPK3 and p-MLKL protein significantly increased(P<0.01).Compared with the model group,the treatment groups showed an increased number of nucleus pulposus cells with orderly arrangement and more extracellular matrix,the ultrastructural damage of the cell membrane,organelle and nucleus in nucleus pulposus cells was partially restored under transmission electron microscopy,the positive expressions of Aggrecan and Collagen Ⅱ significantly increased in Bushen Huoxue Prescription medium-and high-dosage groups and the ibuprofen group(P<0.05,P<0.01),while the expressions of p-RIPK1,p-RIPK3 and p-MLKL protein significantly decreased(P<0.05,P<0.01).Conclusion Bushen Huoxue Prescription may delay intervertebral disc degeneration of the model rabbit by inhibiting the expressions of p-RIPK1,p-RIPK3 and p-MLKL protein in nucleus pulposus cells,and promoting the generation of extracellular matrix components Aggrecan and Collagen Ⅱ.
7.Exploration on the Effect of Bushen Huoxue Prescription on Necroptosis in Human Nucleus Pulposus Cells Based on RIPK1/RIPK3/MLKL Pathway
Wei PENG ; Liguo ZHU ; Xunlu YIN ; Kexin YANG ; Jie LUO ; Minshan FENG ; Jie YU ; Linghui LI ; Jiawen ZHAN ; Tao HAN ; Long LIANG ; Mingyi LUO ; Dian ZHANG
Chinese Journal of Information on Traditional Chinese Medicine 2025;32(3):69-75
Objective To observe the effects of Bushen Huoxue Prescription on pressure-induced necroptosis in human nucleus pulposus cells and the expressions of RIPK1/RIPK3/MLKL pathway;To explore its potential mechanism in delaying intervertebral disc degeneration.Methods Human primary nucleus pulposus cells were cultured in vitro,and a model of nucleus pulposus cell degeneration was established using continuous load pressure method.After modeling,the nucleus pulposus cells were divided into model group,Bushen Huoxue Prescription group and inhibitor group,blank serum,Bushen Huoxue Prescription containing serum and necroptotic apoptosis inhibitor(Nec-1)intervention were administered,respectively.Normal group nucleus pulposus cells were cultured routinely.AO/EB fluorescence dual staining method was used for detecting cell apoptosis,flow cytometry was used to detect the necroptosis rate of nucleus pulposus cells,Western blot was used to detect the protein expressions of p-receptor interacting protein kinase(RIPK)1,p-RIPK3 and p-mixed lineage kinase domain like protein(MLKL),RT-qPCR was used to detect the mRNA expressions of RIPK1,RIPK3 and MLKL.Results Compared with the normal group,the model group showed more red fluorescence under AO/EB staining of nucleus pulposus cells,which were round and condensed,the necroptosis rate of nucleus pulposus cells increased(P<0.05),the protein expressions of p-RIPK1,p-RIPK3 and p-MLKL increased(P<0.05,P<0.01),while there was no significant difference in RIPK1 mRNA expression(P>0.05),and RIPK3 and MLKL mRNA expression increased(P<0.01).Compared with the model group,Bushen Huoxue Prescription group and the inhibitor group had less red condensed chromatin in the nucleus pulposus cells,Bushen Huoxue Prescription group had a lower rate of necroptosis(P<0.05),while the inhibitor group showed a decreasing trend in necroptosis rate(P>0.05),the protein expressions of p-RIPK1,p-RIPK3 and p-MLKL decreased in Bushen Huoxue Prescription group and the inhibitor group(P<0.05,P<0.01),while there was no significant difference in RIPK1 mRNA expression(P>0.05),and RIPK3 and MLKL mRNA expressions decreased(P<0.01).Conclusion Bushen Huoxue Prescription can alleviate pressure-induced damage to nucleus pulposus cells and inhibit necroptosis,thereby slowing the progression of intervertebral disc degeneration.Its mechanism may be related to the RIPK1/RIPK3/MLKL pathway mediated necroptosis.
8.Differences in cytokines expression between mild and severe infant cases infected with respiratory syncytial virus
Guangyu XUE ; Yuting HU ; Kexin ZONG ; Qin LUO ; Shengnan YANG ; Miao FENG ; Xiaoyu YI ; Zhiqiang XIA ; Chen GAO ; Haijun DU ; Ying LI ; Ying CHEN ; Feng HE ; Yajuan WANG ; Yingli QU ; Jin CAO ; Wenyan TIAN ; Qinqin SONG ; Hailan YAO ; Jun HAN
Chinese Journal of Experimental and Clinical Virology 2025;39(3):370-377
Objective:To analyze the clinical characteristics and cytokines expression characteristics in infants with mild and severe respiratory syncytial virus (RSV) infection.Methods:From May 2023 to December 2023, plasma samples and clinical information were collected from 16 infants with RSV infection and 14 control infants. Cytek Aurora flow cytometry (Cytek, America) and Enzyme linked immunosorbent assay (ELISA) were used to detect the expression levels of 25 cytokines after mild and severe RSV infection.Results:Cough and nasal obstruction were the main clinical manifestations in infants with mild RSV infection, accompanied by polypnea, wheezing and other symptoms. The main symptoms of severe RSV infection were cough and rales, accompanied by fever and polypnea. In comparison with the control group, the expression levels of IL-2, IL-4, IL-5, IL-6, IL-9, IL-13, IL-22, TNF-α, IFN-α, IFN-β, MIP-1β, I-TAC, ENA-78, GROα, Eotaxin, and MCP-1 in the RSV infection group all exhibited an upregulation trend. Both IP-10 and MIP-3α demonstrated a downward trend in the RSV infection group; however, there was no statistically significant difference ( P>0.05). The levels of IL-10, IFN-γ, MIP-1α, and IL-8 in the RSV infection group were significantly higher than those in the control group, whereas the levels of MIG, TARC, and RANTES in the RSV infection group were significantly lower than those in the control group ( P<0.05). The levels of IL-2, IL-4, IL-5, IL-6, IL-9, IL-10, IL-13, IL-22, IFN-β, IFN-γ, TNF-α, IL-8, I-TAC, MIP-1β, Eotaxin, and MCP-1 in the mild RSV infection group were significantly higher than those in the severe RSV infection group ( P>0.05). Among these, the levels of MIG, RANTES, TARC, MIP-3α, and ENA-78 in the mild infection group were all lower than those in the severe infection group. The expressions of ENA-78 and MIP-1α in the severe infection group were significantly higher than those in the mild infection group and also higher than those in the control group. There was no significant difference in IP-10 and GROα between the mild and severe RSV infection groups ( P>0.05). Conclusions:The differences in clinical features and cytokines between infants with mild and severe RSV infection provide important data support for the prevention and treatment of RSV infection in infants.
9.Exploration on the Effects of Bushen Huoxue Prescription on Rabbit Intervertebral Disc Degeneration Based on the RIPK1/RIPK3/MLKL Signaling Pathway
Wei PENG ; Liguo ZHU ; Xunlu YIN ; Jie LUO ; Kexin YANG ; Minshan FENG ; Jie YU ; Long LIANG ; Linghui LI ; Jiawen ZHAN ; Tao HAN ; Mingyi LUO ; Dian ZHANG
Chinese Journal of Information on Traditional Chinese Medicine 2025;32(2):71-77
Objective To observe the effects of Bushen Huoxue Prescription on the pathway related to necroptosis of nucleus pulposus cells in a model rabbit of intervertebral disc degeneration;To explore its mechanisms in delaying intervertebral disc degeneration.Methods A intervertebral disc degeneration rabbit model was established using the spinal instability method.Totally 40 model rabbits were randomly divided into model group,ibuprofen group and Bushen Huoxue Prescription low-,medium-and high-dosage groups.Additionally,a normal control group and a sham-operation group were set up,with 8 rabbits in each group.Each treatment groups received the corresponding drugs via gavage for two consecutive weeks.HE staining was used to observe morphology of nucleus pulposus tissue,transmission electron microscopy was used to observe ultrastructure in nucleus pulposus cells,immunohistochemical staining was used to assess the expressions of Aggrecan and Collagen Ⅱ in nucleus pulposus tissue,Western blot was used to detect the expressions of p-RIPK1,p-RIPK3 and p-MLKL protein in nucleus pulposus tissue.Results Compared with the sham-operation group,the model group showed a significant decrease in nucleus pulposus cells,disordered cell arrangement,reduced extracellular matrix,interrupted cell membrane continuity under transmission electron microscopy,organelle swelling,nuclear membrane disruption,partial chromatin loss,and positive expression of Aggrecan and Collagen Ⅱ in nucleus pulposus tissue decreased(P<0.01),while the expressions of p-RIPK1,p-RIPK3 and p-MLKL protein significantly increased(P<0.01).Compared with the model group,the treatment groups showed an increased number of nucleus pulposus cells with orderly arrangement and more extracellular matrix,the ultrastructural damage of the cell membrane,organelle and nucleus in nucleus pulposus cells was partially restored under transmission electron microscopy,the positive expressions of Aggrecan and Collagen Ⅱ significantly increased in Bushen Huoxue Prescription medium-and high-dosage groups and the ibuprofen group(P<0.05,P<0.01),while the expressions of p-RIPK1,p-RIPK3 and p-MLKL protein significantly decreased(P<0.05,P<0.01).Conclusion Bushen Huoxue Prescription may delay intervertebral disc degeneration of the model rabbit by inhibiting the expressions of p-RIPK1,p-RIPK3 and p-MLKL protein in nucleus pulposus cells,and promoting the generation of extracellular matrix components Aggrecan and Collagen Ⅱ.
10.Exploration on the Effect of Bushen Huoxue Prescription on Necroptosis in Human Nucleus Pulposus Cells Based on RIPK1/RIPK3/MLKL Pathway
Wei PENG ; Liguo ZHU ; Xunlu YIN ; Kexin YANG ; Jie LUO ; Minshan FENG ; Jie YU ; Linghui LI ; Jiawen ZHAN ; Tao HAN ; Long LIANG ; Mingyi LUO ; Dian ZHANG
Chinese Journal of Information on Traditional Chinese Medicine 2025;32(3):69-75
Objective To observe the effects of Bushen Huoxue Prescription on pressure-induced necroptosis in human nucleus pulposus cells and the expressions of RIPK1/RIPK3/MLKL pathway;To explore its potential mechanism in delaying intervertebral disc degeneration.Methods Human primary nucleus pulposus cells were cultured in vitro,and a model of nucleus pulposus cell degeneration was established using continuous load pressure method.After modeling,the nucleus pulposus cells were divided into model group,Bushen Huoxue Prescription group and inhibitor group,blank serum,Bushen Huoxue Prescription containing serum and necroptotic apoptosis inhibitor(Nec-1)intervention were administered,respectively.Normal group nucleus pulposus cells were cultured routinely.AO/EB fluorescence dual staining method was used for detecting cell apoptosis,flow cytometry was used to detect the necroptosis rate of nucleus pulposus cells,Western blot was used to detect the protein expressions of p-receptor interacting protein kinase(RIPK)1,p-RIPK3 and p-mixed lineage kinase domain like protein(MLKL),RT-qPCR was used to detect the mRNA expressions of RIPK1,RIPK3 and MLKL.Results Compared with the normal group,the model group showed more red fluorescence under AO/EB staining of nucleus pulposus cells,which were round and condensed,the necroptosis rate of nucleus pulposus cells increased(P<0.05),the protein expressions of p-RIPK1,p-RIPK3 and p-MLKL increased(P<0.05,P<0.01),while there was no significant difference in RIPK1 mRNA expression(P>0.05),and RIPK3 and MLKL mRNA expression increased(P<0.01).Compared with the model group,Bushen Huoxue Prescription group and the inhibitor group had less red condensed chromatin in the nucleus pulposus cells,Bushen Huoxue Prescription group had a lower rate of necroptosis(P<0.05),while the inhibitor group showed a decreasing trend in necroptosis rate(P>0.05),the protein expressions of p-RIPK1,p-RIPK3 and p-MLKL decreased in Bushen Huoxue Prescription group and the inhibitor group(P<0.05,P<0.01),while there was no significant difference in RIPK1 mRNA expression(P>0.05),and RIPK3 and MLKL mRNA expressions decreased(P<0.01).Conclusion Bushen Huoxue Prescription can alleviate pressure-induced damage to nucleus pulposus cells and inhibit necroptosis,thereby slowing the progression of intervertebral disc degeneration.Its mechanism may be related to the RIPK1/RIPK3/MLKL pathway mediated necroptosis.

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