1.The multi-center mid-term clinical outcomes of combined complete preservation of chordal structure mitral valve replacement with total anatomic complete arterial myocardial revascularization for coronary patients with moderate-to-severe or severe ischemic mitral regurgitation
Ke GUO ; Xujun CHEN ; Baoshi ZHENG ; Chao SHI ; Keli HUANG ; Yong CAO ; Chengquan LIAO ; Jingwei CHEN ; Yu LIN ; Chengxin LIU ; Quansheng CAO ; Lin SHEN ; Zhendong WANG
Chinese Journal of Surgery 2025;63(1):58-67
Objective:To evaluate the clinical outcomes of combined complete preservation of chordal structure mitral valve replacement (C-MVR) with total anatomical arterial myocardial revascularization (TACR) in coronary patients with moderate-to-severe or severe ischemic mitral regurgitation (IMR).Methods:This is a retrospective multi-center case series study. Data were retrospectively collected from 127 patients with coronary artery disease with moderate to severe or severe IMR who received TACR with C-MVR from July 2015 to April 2024 in 13 hospitals in China. There were 90 males and 37 females, aged (56.5±10.7) years (range: 33 to 74 years). Perioperative data and follow-up data including left ventricular ejection fraction, left ventricular end-diastolic diameter, and patency rate of arterial grafts of patients were collected. Comparisons were made using paired sample t-test or χ2 test. Results:In this cohort of 127 patients, 67 underwent concurrent tricuspid valve repair. During surgery, 113 grafts of the left internal mammary artery (LIMA), 127 grafts of the left radial artery, 80 grafts of the right radial artery, and 110 grafts of the right internal mammary artery (RIMA) were harvested. The number of the distal anastomosis was 4.2±0.4 (range: 3 to 5). The aortic cross-clamp time and cardiopulmonary bypass time were (97.5±23.4) minutes (range: 90 to 161 minutes) and (145.4±19.2) minutes (range: 101 to 210 minutes), respectively. There was one operative death. Intraoperative placement of an intra-aortic balloon pump was performed in 21 patients to improve the left ventricular ejection. No sternal ischemic occurred. All patients completed follow-up, with a mean follow-up period of (64.3±7.5) months (range: 4 to 110 months). No major cerebrovascular events occurred during the follow-up period, and all patients survived. Left ventricular ejection fraction improved postoperatively (55.0%±5.3% vs. 41.0%±15.3%, t=17.23, P<0.01). The proportion of patients with New York Heart Association functional class ≤2 increased postoperatively (23.6% (30/127) vs. 87.3% (110/126), χ2=103.77, P<0.01). The proportion of patients with Canadian Cardiovascular Society Angina Classification ≤3 decreased postoperatively (4.8% (6/126) vs. 78.7% (100/127), χ2=142.19, P<0.01). The left ventricular end-diastolic diameter decreased postoperatively ((5.70±4.50) cm vs. (6.10±0.23) cm, t=12.15, P<0.01). Coronary multi-detector computed tomography angiography (MDCTA) follow-up was conducted for (60.5±11.7) months (range: 6 to 109 months) postoperatively. MDCTA confirmed the patency rates of the grafts: 96.4% (108/112) for the LIMA grafts, 88.9% (112/126) for the left radial artery grafts, 93.7% (74/79) for the right radial artery grafts, and 90.9% (100/110) for the free RIMA grafts. No significant differences in graft patency rates were observed between the arterial grafts ( χ2=5.24, P=0.155). Conclusion:The results of this multi-centre study demonstrate satisfactory mid-term results of C-MVR with TACR for the treatment of coronary artery disease with moderate to severe or severe IMR.
2.The multi-center mid-term clinical outcomes of combined complete preservation of chordal structure mitral valve replacement with total anatomic complete arterial myocardial revascularization for coronary patients with moderate-to-severe or severe ischemic mitral regurgitation
Ke GUO ; Xujun CHEN ; Baoshi ZHENG ; Chao SHI ; Keli HUANG ; Yong CAO ; Chengquan LIAO ; Jingwei CHEN ; Yu LIN ; Chengxin LIU ; Quansheng CAO ; Lin SHEN ; Zhendong WANG
Chinese Journal of Surgery 2025;63(1):58-67
Objective:To evaluate the clinical outcomes of combined complete preservation of chordal structure mitral valve replacement (C-MVR) with total anatomical arterial myocardial revascularization (TACR) in coronary patients with moderate-to-severe or severe ischemic mitral regurgitation (IMR).Methods:This is a retrospective multi-center case series study. Data were retrospectively collected from 127 patients with coronary artery disease with moderate to severe or severe IMR who received TACR with C-MVR from July 2015 to April 2024 in 13 hospitals in China. There were 90 males and 37 females, aged (56.5±10.7) years (range: 33 to 74 years). Perioperative data and follow-up data including left ventricular ejection fraction, left ventricular end-diastolic diameter, and patency rate of arterial grafts of patients were collected. Comparisons were made using paired sample t-test or χ2 test. Results:In this cohort of 127 patients, 67 underwent concurrent tricuspid valve repair. During surgery, 113 grafts of the left internal mammary artery (LIMA), 127 grafts of the left radial artery, 80 grafts of the right radial artery, and 110 grafts of the right internal mammary artery (RIMA) were harvested. The number of the distal anastomosis was 4.2±0.4 (range: 3 to 5). The aortic cross-clamp time and cardiopulmonary bypass time were (97.5±23.4) minutes (range: 90 to 161 minutes) and (145.4±19.2) minutes (range: 101 to 210 minutes), respectively. There was one operative death. Intraoperative placement of an intra-aortic balloon pump was performed in 21 patients to improve the left ventricular ejection. No sternal ischemic occurred. All patients completed follow-up, with a mean follow-up period of (64.3±7.5) months (range: 4 to 110 months). No major cerebrovascular events occurred during the follow-up period, and all patients survived. Left ventricular ejection fraction improved postoperatively (55.0%±5.3% vs. 41.0%±15.3%, t=17.23, P<0.01). The proportion of patients with New York Heart Association functional class ≤2 increased postoperatively (23.6% (30/127) vs. 87.3% (110/126), χ2=103.77, P<0.01). The proportion of patients with Canadian Cardiovascular Society Angina Classification ≤3 decreased postoperatively (4.8% (6/126) vs. 78.7% (100/127), χ2=142.19, P<0.01). The left ventricular end-diastolic diameter decreased postoperatively ((5.70±4.50) cm vs. (6.10±0.23) cm, t=12.15, P<0.01). Coronary multi-detector computed tomography angiography (MDCTA) follow-up was conducted for (60.5±11.7) months (range: 6 to 109 months) postoperatively. MDCTA confirmed the patency rates of the grafts: 96.4% (108/112) for the LIMA grafts, 88.9% (112/126) for the left radial artery grafts, 93.7% (74/79) for the right radial artery grafts, and 90.9% (100/110) for the free RIMA grafts. No significant differences in graft patency rates were observed between the arterial grafts ( χ2=5.24, P=0.155). Conclusion:The results of this multi-centre study demonstrate satisfactory mid-term results of C-MVR with TACR for the treatment of coronary artery disease with moderate to severe or severe IMR.
3.Efficacy of Fufang Lingjiao Jiangya Pills with Different Proportions of Goat Horn Replacing Antelope Horn on Spontaneous Hypertensive Rats
Tengjian WANG ; Wanlu ZHAO ; Yang YU ; Yan LIU ; Kun CAO ; Zheyuan LIN ; Yue WU ; Lilan LUO ; Weizhi LAI ; Zhaohuan LOU ; Qiaoyan ZHANG ; Quanlong ZHANG ; Luping QIN
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(9):68-78
ObjectiveTo investigate the optimal ratio of goat horn replacing antelope horn in Fufang Lingjiao Jiangya pills and the blood pressure-lowering mechanism of this medicine. MethodsThe blood pressure-lowering efficacy of Fufang Lingjiao Jiangya pills with varying proportions of goat horn replacing antelope horn was evaluated on spontaneous hypertensive rats (SHR). In this experiment, 50 SHR rats were randomly grouped as follows: model (n=8), captopril (0.01 g·kg-1) (n=6), low-dose blank Fufang Lingjiao Jiangya pills (0.342 g·kg-1) (n=6), high-dose blank Fufang Lingjiao Jiangya pills (0.684 g·kg-1) (n=6), low-dose antelope horn-containing Fufang Lingjiao Jiangya pills (0.378 g·kg-1) (n=6), high-dose antelope horn-containing Fufang Lingjiao Jiangya pills (0.756 g·kg-1) (n=6), low-dose goat horn-containing Fufang Lingjiao Jiangya pills (0.378 g·kg-1) (n=6), and high-dose goat horn-containing Fufang Lingjiao Jiangya pills (0.756 g·kg-1) (n=6). Additionally, 8 WKY rats were used as the normal group. Drugs were administered by gavage for 4 weeks while an equal volume of distilled water was administered for the normal and model groups. Blood pressure was measured before administration, 3 h post administration, and biweekly thereafter. In the experiment for Fufang Lingjiao Jiangya pills with goat horn replacing antelope horn in different proportions, 48 SHR rats were randomly grouped as follows: model, blank Fufang Lingjiao Jiangya pills (0.684 g·kg-1), antelope horn-containing Fufang Lingjiao Jiangya pills (0.756 g·kg-1), 2× goat horn-containing Fufang Lingjiao Jiangya pills (0.824 g·kg-1), 4× goat horn Fufang Lingjiao Jiangya pills (0.969 g·kg-1), and 6× goat horn Fufang Lingjiao Jiangya pills (1.112 g·kg-1). The normal group included 8 WKY rats, and the normal group and model group received an equal volume of distilled water. The treatment lasted for 2 weeks, and blood pressure was recorded at various time points (pre-administration, 3 h post administration, and on days 4, 7, 10, and 14 of administration). Serum levels of angiotensin-converting enzyme (ACE), angiotensin Ⅱ(Ang Ⅱ), renin, and interleukin-6 (IL-6) were measured by enzyme-linked immunosorbent assay. Histopathological changes in the heart, kidney, and thoracic aorta were observed by hematoxylin-eosin staining. The protein levels of ACE2, angiotensin Ⅱ type 1 receptor (AT1R), and angiotensinogen (AGT) in the kidney tissue were determined by Western blot, while the expression of nuclear factor (NF)-κB p65 and Toll-like receptor 4 (TLR4) in the thoracic aorta tissue was assessed by immunohistochemistry. ResultsCompared with the model group, all treatment groups showed lowered blood pressure (P<0.05, P<0.01), and the 6× goat horn-containing Fufang Lingjiao Jiangya pills group showed consistent blood pressure-lowering effect with the antelope horn-containing Fufang Lingjiao Jiangya pills group. Compared with the normal group, the model group showed elevated serum levels of ACE, Ang Ⅱ, renin, and IL-6, while the elevations were declined in the Fufang Lingjiao Jiangya pills groups (P<0.05, P<0.01). Pathological changes in the heart, kidney, and thoracic aorta were alleviated in all the treatment groups, with the 6× goat horn- and antelope horn-containing Fufang Lingjiao Jiangya pills groups exhibited the best effect. Western blot and immunohistochemistry results showed that all the treatment groups exhibited down-regulated protein levels of AT1R, AGT, NF-κB p65, and TLR4 and up-regulated protein levels of ACE2 (P<0.05, P<0.01) compared with model group, with the 6×goat horn- and antelope horn-containing Fufang Lingjiao Jiangya pills groups showcasing the best effect. ConclusionReplacing antelope horn with 6×goat horn in Fufang Lingjiao Jiangya pills can achieve consistent blood pressure-lowering effect with the original prescription. The prescription may exert the effect by inhibiting the renin-angiotensin-aldosterone system (RAAS) and TLR4/NF-κB signaling pathways.
4.Mechanism of Traditional Chinese Medicine Regulating JAK/STAT Signaling Pathway to Intervene in Lung Cancer: A Review
Jiarui CAO ; Bo FENG ; Chunzheng MA ; Weixia CHEN ; Jiangfan YU ; Shasha CAO ; Zhenyu ZHANG ; Wenhui OUYANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(9):265-276
Lung cancer is the malignant tumor with the highest incidence and mortality rates globally. Current treatment methods for lung cancer primarily include surgery, chemotherapy, targeted therapy, and immunotherapy. However, the main limitations of these treatments are their side effects, the drug resistance, and the economic burden they impose. As a critical cancer pathway, the Janus kinase (JAK)/signal transducer and activator of transcription (STAT) signaling pathway regulates tumor occurrence and development through multiple mechanisms by influencing various downstream targets. Consequently, the JAK/STAT signaling pathway offers a promising avenue for lung cancer treatment research. Numerous studies have demonstrated that the JAK/STAT signaling pathway plays a key role in the proliferation and growth of lung cancer cells, angiogenesis, epithelial-mesenchymal transition (EMT), metabolic alterations, remodeling of the immune microenvironment, and the development of treatment resistance. Traditional Chinese medicine (TCM) has garnered increasing attention due to its minimal side effects, low economic burden, and its potential to enhance efficacy and reduce toxicity when used in conjunction with Western medicine. In addition to traditional Chinese medicine compounds, a growing number of Chinese medicine monomers have come into the spotlight because of their more targeted effects. Numerous studies investigating the regulation of the JAK/STAT signaling pathway by TCM in the treatment of lung cancer have demonstrated that TCM can inhibit the proliferation and invasion of lung cancer cells, tumor angiogenesis, and EMT, improve the inflammatory and immunosuppressive microenvironments, and enhance treatment sensitivity by intervening in the JAK/STAT signaling pathway, thereby impeding the progression of lung cancer. In recent years, the research on the regulation of this pathway by TCM in the treatment of lung cancer has been updated rapidly. However, the summary of these studies has not been updated in time. This review summarizes and reflects on the recent research findings regarding the regulation of the JAK/STAT signaling pathway by TCM to intervene in lung cancer from three aspects, introducing the JAK/STAT pathway, elaborating the mechanism of this pathway in lung cancer, and exploring the intervention of TCM in the treatment of lung cancer through this pathway, to provide more reference for the treatment of lung cancer in the future.
5.Alleviation of Ulcerative Colitis by Shaoyaotang via Inhibiting Glycolysis Through SIRT6/HIF-1α Pathway
Yiling XIA ; Hui CAO ; Dongsheng WU ; Bo ZOU ; Erle LIU ; Yiwen WANG ; Shaijin JIANG ; Yiqian YU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(11):10-19
ObjectiveTo investigate the role of silent information regulatory protein (SIRT6)/hypoxia-inducible factor-1α (HIF-1α) pathway in regulating the reprogramming of glucose metabolism in ulcerative colitis (UC) and the mechanism of intervention of Shaoyaotang. MethodsForty-eight c57bL/6 mice were randomly divided into a blank group, a model group, a Mesalazine group (0.42 g·kg-1), a Shaoyaotang group (31.08 g·kg-1), an inhibitor group (OSS-128167, 50 mg·kg-1), and an inhibitor + Shaoyaotang group (50 mg·kg-1 OSS-128167 + 31.08 g·kg-1 Shaoyaotang). A UC model was established by the administration of 2.5% dextran sulfate sodium (DSS) solution for mice in other groups for 7 d, except for the blank group. The mice in each group were treated with saline, Mesalazine, Shaoyaotang, inhibitor, and inhibitor + Shaoyaotang, respectively, for 7 d. The mice were necropsied 24 h after the last administration of the drug. The blood was collected from the orbital region, and colon tissue was taken. Hematoxylin-eosin (HE) staining was used to observe the pathological changes in colon tissue. Enzyme-linked immunosorbent assay (ELISA) was employed to detect serum interleukin (IL)-10, IL-17, and IL-6 levels. A biochemical method was used to detect glucose and lactate dehydrogenase A (LDHA) levels. Immunohistochemistry (IHC) was employed to detect IL-22 and transforming growth factor-β1 (TGF-β1) levels in colon tissue, and Western blot and real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) were used to detect relative protein and mRNA expressions of SIRT6, HIF-1α, and LDHA. ResultsCompared with those of the blank group, disease activity index (DAI) scores of mice in the model group and inhibitor group were significantly increased (P<0.01). The length of colon tissue was significantly shortened, and colon tissue was congested and eroded. The pathohistological scores were significantly increased (P<0.01). The levels of serum inflammatory factors IL-17 and IL-6 were significantly elevated, and the levels of IL-10 were significantly decreased (P<0.01). The protein expressions of IL-22 and TGF-β1 were significantly reduced in colon tissue (P<0.01). The relative protein and mRNA expressions of SIRT6 were significantly decreased (P<0.01), and the relative protein and mRNA expressions of HIF-1α and LDHA and the contents of glucose and lactate were significantly elevated (P<0.01). The level of inflammation in the colon of the mice in the inhibitor group was more severe than that in the model group (P<0.01). Compared with the model group, the Mesalazine group, the Shaoyaotang group, and the inhibitor + Shaoyaotang group showed reduced colonic injury, significant decrease in serum IL-17 and IL-6, significant increase in IL-10 (P<0.01), significant increase in the protein expressions of IL-22 and TGF-β1 in colon tissue (P<0.01), significant increase in the protein expressions of SIRT6 and the relative mRNA expressions (P<0.01), and significant reduction in the protein expressions of HIF-1α and LDHA, the relative mRNA expressions, and the contents of glucose and lactate (P<0.01). Compared with those in the Shaoyaotang group, the serum IL-17 and IL-6 were significantly increased, and IL-10 was significantly decreased in the inhibitor + Shaoyaotang group (P<0.01). The protein expressions of IL-22 and TGF-β1 in colon tissue were significantly decreased (P<0.01). The expressions of SIRT6 protein and the relative mRNA expressions were significantly decreased (P<0.01). The protein expressions of HIF-1α and LDHA, the relative mRNA expressions, and the contents of glucose and lactate were significantly elevated (P<0.01). However, the difference between the Shaoyaotang group and the Mesalazine group was not significant. ConclusionShaoyaotang can effectively treat DSS-induced mice with UC through the SIRT6/HIF-1α pathway, and its mechanism of action may be related to the regulation of the SIRT6/HIF-1α pathway and glucose metabolism reprogramming and the inhibition of glycolysis.
6.Alleviation of Ulcerative Colitis by Shaoyaotang via Inhibiting Glycolysis Through SIRT6/HIF-1α Pathway
Yiling XIA ; Hui CAO ; Dongsheng WU ; Bo ZOU ; Erle LIU ; Yiwen WANG ; Shaijin JIANG ; Yiqian YU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(11):10-19
ObjectiveTo investigate the role of silent information regulatory protein (SIRT6)/hypoxia-inducible factor-1α (HIF-1α) pathway in regulating the reprogramming of glucose metabolism in ulcerative colitis (UC) and the mechanism of intervention of Shaoyaotang. MethodsForty-eight c57bL/6 mice were randomly divided into a blank group, a model group, a Mesalazine group (0.42 g·kg-1), a Shaoyaotang group (31.08 g·kg-1), an inhibitor group (OSS-128167, 50 mg·kg-1), and an inhibitor + Shaoyaotang group (50 mg·kg-1 OSS-128167 + 31.08 g·kg-1 Shaoyaotang). A UC model was established by the administration of 2.5% dextran sulfate sodium (DSS) solution for mice in other groups for 7 d, except for the blank group. The mice in each group were treated with saline, Mesalazine, Shaoyaotang, inhibitor, and inhibitor + Shaoyaotang, respectively, for 7 d. The mice were necropsied 24 h after the last administration of the drug. The blood was collected from the orbital region, and colon tissue was taken. Hematoxylin-eosin (HE) staining was used to observe the pathological changes in colon tissue. Enzyme-linked immunosorbent assay (ELISA) was employed to detect serum interleukin (IL)-10, IL-17, and IL-6 levels. A biochemical method was used to detect glucose and lactate dehydrogenase A (LDHA) levels. Immunohistochemistry (IHC) was employed to detect IL-22 and transforming growth factor-β1 (TGF-β1) levels in colon tissue, and Western blot and real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) were used to detect relative protein and mRNA expressions of SIRT6, HIF-1α, and LDHA. ResultsCompared with those of the blank group, disease activity index (DAI) scores of mice in the model group and inhibitor group were significantly increased (P<0.01). The length of colon tissue was significantly shortened, and colon tissue was congested and eroded. The pathohistological scores were significantly increased (P<0.01). The levels of serum inflammatory factors IL-17 and IL-6 were significantly elevated, and the levels of IL-10 were significantly decreased (P<0.01). The protein expressions of IL-22 and TGF-β1 were significantly reduced in colon tissue (P<0.01). The relative protein and mRNA expressions of SIRT6 were significantly decreased (P<0.01), and the relative protein and mRNA expressions of HIF-1α and LDHA and the contents of glucose and lactate were significantly elevated (P<0.01). The level of inflammation in the colon of the mice in the inhibitor group was more severe than that in the model group (P<0.01). Compared with the model group, the Mesalazine group, the Shaoyaotang group, and the inhibitor + Shaoyaotang group showed reduced colonic injury, significant decrease in serum IL-17 and IL-6, significant increase in IL-10 (P<0.01), significant increase in the protein expressions of IL-22 and TGF-β1 in colon tissue (P<0.01), significant increase in the protein expressions of SIRT6 and the relative mRNA expressions (P<0.01), and significant reduction in the protein expressions of HIF-1α and LDHA, the relative mRNA expressions, and the contents of glucose and lactate (P<0.01). Compared with those in the Shaoyaotang group, the serum IL-17 and IL-6 were significantly increased, and IL-10 was significantly decreased in the inhibitor + Shaoyaotang group (P<0.01). The protein expressions of IL-22 and TGF-β1 in colon tissue were significantly decreased (P<0.01). The expressions of SIRT6 protein and the relative mRNA expressions were significantly decreased (P<0.01). The protein expressions of HIF-1α and LDHA, the relative mRNA expressions, and the contents of glucose and lactate were significantly elevated (P<0.01). However, the difference between the Shaoyaotang group and the Mesalazine group was not significant. ConclusionShaoyaotang can effectively treat DSS-induced mice with UC through the SIRT6/HIF-1α pathway, and its mechanism of action may be related to the regulation of the SIRT6/HIF-1α pathway and glucose metabolism reprogramming and the inhibition of glycolysis.
7.Effect of Zuoguiwan on Development of Skin Barrier in Neonatal Rat Model of Congenital Kidney Deficiency Based on Intercellular Connections
He YU ; Min XIAO ; Xiaocui JIANG ; Min ZHAO ; Yinjuan LYU ; Jian GONG ; Jigang CAO
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(13):11-18
ObjectiveTo study the effect of Zuoguiwan on the development of skin barrier in the neonatal rat model of congenital kidney deficiency and unveil the underlying mechanism. MethodsSixty rats were paired in a female-to-male ratio of 2∶1, and the pregnant rats were assigned into control, congenital kidney deficiency, and low- and high-dose (2 and 8 g·kg-1, respectively) Zuoguiwan groups. The pregnant rats in other groups except the control group were exposed to chronic unpredictable mild stress for the modeling of congenital kidney deficiency. The rats in the control group and congenital kidney deficiency group were administrated with normal saline by gavage, and those in Zuoguiwan groups with Zuoguiwan suspension by gavage from day 1 of pregnancy. The serum level of interleukin-6 (IL-6) in the neonatal rats on the day of birth was determined by enzyme-linked immunosorbent assay. The full-thickness skin of neonatal rats on the day of birth was removed from the same position on the back and stained with hematoxylin-eosin for observation of histopathological changes, measurement of skin thickness, and counting of hair follicles. Terminal deoxynucleotidyl transferase-mediated nick end labeling was used to detect the apoptosis of skin tissue cells. The expression of interleukin-6 receptor (IL-6R) and interleukin-17A (IL-17A) in the skin tissue was assessed by immunohistochemistry and Western blot, and the expression of occludin, connexin 43 (Cx43), and zonula occludens-1 (ZO-1) in the skin tissue was assessed by immunofluorescence and Western blot. ResultsCompared with those in the control group, the neonatal rats in the congenital kidney deficiency group showed a rise in the serum IL-6 level (P<0.01), decreases in stratum corneum thickness, skin thickness, and number of hair follicles (P<0.01), increases in the expression of IL-6R and IL-17A in the skin tissue (P<0.01) and the number of apoptotic cells (P<0.01), and decreases in the expression of occludin, Cx43, ZO-1 (P<0.05). Compared with the congenital kidney deficiency group, the low- and high-dose Zuoguiwan groups showed declines in serum IL-6 level (P<0.05). The low-dose group showed increased number of hair follicles (P<0.05), and the high-dose group presented thickened stratum corneum (P<0.01), increased number of hair follicles (P<0.01), and down-regulated expression of IL-6R and IL-17A in the skin tissue (P<0.05, P<0.01). Both Zuoguiwan groups showcased decreased number of apoptotic cells (P<0.05, P<0.01), and the high-dose group showed up-regulated expression of occludin, Cx43, and ZO-1 in the skin tissue (P<0.05, P<0.01). ConclusionZuoguiwan can reduce the levels of IL-6 in the serum and IL-6R and IL-17A in the skin tissue and improve the expression of intercellular junction proteins, thereby ameliorating the abnormal development of the skin barrier in the neonatal rat model of congenital kidney deficiency.
8.Shaoyaotang Containing Serum Mediates Fas/FasL Pathway to Inhibit Lipopolysaccharide Induced Inflammation and Apoptosis of Caco-2 Cells
Yuting YANG ; Dongsheng WU ; Hui CAO ; Yu ZHANG ; Nianjia XIE ; Bo ZOU ; Daguang CHEN ; Erle LIU ; Yi LU ; Zhaowen LYU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(13):62-69
ObjectiveTo investigate the effects of different concentrations of Shaoyaotang-containing serum on lipopolysaccharide (LPS)-induced inflammation of human colorectal adenocarcinoma (Caco-2) cells by inhibiting apoptosis via activating the tumor necrosis factor (TNF) receptor superfamily member 6 (Fas)/Fas ligand (FasL) pathway. MethodsCaco-2 cells were allocated into blank, model (LPS, 10 mg·L-1), Shaoyaotang-containing serum (5%, 10%, 15%, 20%), and Fas inhibitor (KR-33493, 20 mmol·L-1) groups. Except the blank group, the other groups were stimulated with 10 mg·L-1 LPS for 24 h for the modeling of inflammation. After successful modeling, the blank, Fas inhibitor, and model groups were treated with blank serum, and the Shaoyaotang-containing serum groups were treated with the serum samples at corresponding concentrations for 24 h. The Fas inhibitor group was subjected to KR-33493 pretreatment for 1 h. Cell proliferation and viability were examined by the cell-counting kit-8 (CCK-8) method. The levels of interleukin (IL)-6, IL-1β, and TNF-α were measured by enzyme-linked immunosorbent assay. Apoptosis was detected by flow cytometry. The protein and mRNA levels of Fas, FasL, cysteinyl aspartate-specific proteinase (Caspase)-3, Caspase-9, B-cell lymphoma 2 (Bcl-2), and Bcl-2-associated X protein (Bax) were determined by Western blot and Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR), respectively. ResultsCompared with the blank group, the model group presented a decrease in cell survival rate (P<0.01). Compared with that in the model group, the cell survival rate showed no significant change in the 5% Shaoyaotang-containing serum group but increased in the 10%, 15%, and 20% Shaoyaotang-containing serum groups (P<0.01). Since there was no statistical difference between the 5% Shaoyaotang-containing serum group and the model group, 10%, 15%, and 20% Shaoyaotang-containing sera were selected for the follow-up study. Compared with the blank group, the model group showed risen levels of IL-6, IL-1β, and TNF-α (P<0.01), an increased apoptosis rate (P<0.01), up-regulated protein and mRNA levels of Fas, FasL, Caspase-3, Caspase-9, and Bax (P<0.01), and down-regulated protein and mRNA levels of Bcl-2 (P<0.01). Compared with the model group, the Fas inhibitor group and the 10%, 15%, and 20% Shaoyaotang-containing serum groups showed declined levels of IL-6, IL-1β, and TNF-α (P<0.01), decreased apoptosis rates (P<0.01), down-regulated protein and mRNA levels of Fas, FasL, Caspase-3, Caspase-9, and Bax (P<0.05, P<0.01), and up-regulated protein and mRNA levels of Bcl-2 (P<0.05, P<0.01). In addition, the 15% and 20% Shaoyaotang-containing serum groups had lower levels of IL-6, IL-1β, and TNF-α (P<0.05, P<0.01), lower apoptosis rates (P<0.05, P<0.01), lower protein and mRNA levels of Fas, FasL, Caspase-3, Caspase-9, and Bax (P<0.05, P<0.01), and higher protein and mRNA levels of Bcl-2 (P<0.05, P<0.01) than the 10% Shaoyaotang-containing serum group. ConclusionThe Shaoyaotang-containing serum can reduce the content of inflammatory factors in Caco-2 cells, down-regulate the protein and mRNA levels of Fas, FasL, Caspase-3, Caspase-9, and Bax, and up-regulate the protein and mRNA levels of Bcl-2 under the intervention of LPS by regulating the Fas/FasL pathway and inhibiting the apoptosis of intestinal epithelial cells in ulcerative colitis.
9.Shaoyaotang Alleviates Damage of Tight Junction Proteins in Caco-2 Cell Model of Inflammation by Regulating RhoA/ROCK Pathway
Nianjia XIE ; Dongsheng WU ; Hui CAO ; Yu ZHANG ; Yuting YANG ; Bo ZOU ; Da ZHAO ; Yi LU ; Mingsheng WU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(13):70-77
ObjectiveTo investigate the protective effect and mechanism of Shaoyaotang (SYD) on the lipopolysaccharide (LPS)-induced damage of tight junction proteins in the human colorectal adenocarcinoma (Caco-2) cell model of inflammation via the Ras homolog gene family member A (RhoA)/Rho-associated coiled-coil forming protein kinase (ROCK) pathway. MethodsCaco-2 cells were grouped as follows: Blank, model (LPS, 10 mg·L-1), SYD-containing serum (10%, 15%, and 20%), and inhibitor (Fasudil, 25 μmol·L-1). After 24 hours of intervention, the cell viability in each group was examined by the cell-counting kit 8 (CCK-8) method. Enzyme-linked immunosorbent assay was employed to determine the levels of endothelin-1 (ET-1), tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and interleukin-6 (IL-6). Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) and Western blot were employed to determine the mRNA and protein levels, respectively, of RhoA, ROCK2, claudin-5, and zonula occludens-1 (ZO-1) in cells of each group. ResultsCompared with the blank group, the model group showcased a marked reduction in the cell viability (P<0.01), elevations in the levels of ET-1, TNF-α, IL-1β, and IL-6 (P<0.01), declines in both mRNA and protein levels of ZO-1 and claudin-5 (P<0.01), and rises in mRNA and protein levels of RhoA and ROCK2 (P<0.01). Compared with the model group, the Shaoyaotang-containing serum (10%, 15%, and 20%) groups had enhanced cell viability (P<0.01), lowered levels of ET-1, TNF-α, IL-1β, and IL-6 (P<0.01), up-regulated mRNA and protein levels of ZO-1 and claudin-5 (P<0.05, P<0.01), and down-regulated mRNA and protein levels of RhoA and ROCK2 (P<0.01). Moreover, the inhibitor group and the 15% and 20% Shaoyaotang-containing serum groups had lower levels of ET-1, TNF-α, IL-1β, and IL-6 (P<0.05, P<0.01), higher mRNA and protein levels of ZO-1 and claudin-5 (P<0.05, P<0.01), and lower mRNA and protein levels of RhoA and ROCK2 (P<0.05, P<0.01) than the 10% Shaoyaotang-containing serum group. ConclusionThe Shaoyaotang-containing serum can lower the levels of LPS-induced increases in levels of inflammatory cytokines and endothelin to ameliorate the damage of tight junction proteins of the Caco-2 cell model of inflammation by regulating the expression of proteins in the RhoA/ROCK pathway.
10.Effect of Zuoguiwan on Development of Skin Barrier in Neonatal Rat Model of Congenital Kidney Deficiency Based on Intercellular Connections
He YU ; Min XIAO ; Xiaocui JIANG ; Min ZHAO ; Yinjuan LYU ; Jian GONG ; Jigang CAO
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(13):11-18
ObjectiveTo study the effect of Zuoguiwan on the development of skin barrier in the neonatal rat model of congenital kidney deficiency and unveil the underlying mechanism. MethodsSixty rats were paired in a female-to-male ratio of 2∶1, and the pregnant rats were assigned into control, congenital kidney deficiency, and low- and high-dose (2 and 8 g·kg-1, respectively) Zuoguiwan groups. The pregnant rats in other groups except the control group were exposed to chronic unpredictable mild stress for the modeling of congenital kidney deficiency. The rats in the control group and congenital kidney deficiency group were administrated with normal saline by gavage, and those in Zuoguiwan groups with Zuoguiwan suspension by gavage from day 1 of pregnancy. The serum level of interleukin-6 (IL-6) in the neonatal rats on the day of birth was determined by enzyme-linked immunosorbent assay. The full-thickness skin of neonatal rats on the day of birth was removed from the same position on the back and stained with hematoxylin-eosin for observation of histopathological changes, measurement of skin thickness, and counting of hair follicles. Terminal deoxynucleotidyl transferase-mediated nick end labeling was used to detect the apoptosis of skin tissue cells. The expression of interleukin-6 receptor (IL-6R) and interleukin-17A (IL-17A) in the skin tissue was assessed by immunohistochemistry and Western blot, and the expression of occludin, connexin 43 (Cx43), and zonula occludens-1 (ZO-1) in the skin tissue was assessed by immunofluorescence and Western blot. ResultsCompared with those in the control group, the neonatal rats in the congenital kidney deficiency group showed a rise in the serum IL-6 level (P<0.01), decreases in stratum corneum thickness, skin thickness, and number of hair follicles (P<0.01), increases in the expression of IL-6R and IL-17A in the skin tissue (P<0.01) and the number of apoptotic cells (P<0.01), and decreases in the expression of occludin, Cx43, ZO-1 (P<0.05). Compared with the congenital kidney deficiency group, the low- and high-dose Zuoguiwan groups showed declines in serum IL-6 level (P<0.05). The low-dose group showed increased number of hair follicles (P<0.05), and the high-dose group presented thickened stratum corneum (P<0.01), increased number of hair follicles (P<0.01), and down-regulated expression of IL-6R and IL-17A in the skin tissue (P<0.05, P<0.01). Both Zuoguiwan groups showcased decreased number of apoptotic cells (P<0.05, P<0.01), and the high-dose group showed up-regulated expression of occludin, Cx43, and ZO-1 in the skin tissue (P<0.05, P<0.01). ConclusionZuoguiwan can reduce the levels of IL-6 in the serum and IL-6R and IL-17A in the skin tissue and improve the expression of intercellular junction proteins, thereby ameliorating the abnormal development of the skin barrier in the neonatal rat model of congenital kidney deficiency.

Result Analysis
Print
Save
E-mail