1.Mechanism of Bushen huoxue formula in improving pregnancy outcomes in mice with antiphospholipid antibody-related recurrent spontaneous abortion
Yongmei HAN ; Tianwei CUI ; Jundan XIE ; Xinyu HE ; Yuting GONG ; Manman WANG
China Pharmacy 2026;37(11):1408-1415
OBJECTIVE To investigate the mechanism of Bushen huoxue formula in improving pregnancy outcomes in mice with antiphospholipid antibody-related recurrent spontaneous abortion (aPL-RSA). METHODS SPF female BALB/c mice were randomly divided into normal control group, adjuvant control group, model group, and Bushen huoxue formula group. The aPL-RSA mouse model was established by immunization with β 2 glycoproteinⅠcombined with Freund’s adjuvant. From gestational day 0, the Bushen huoxue formula group was administered 1.653 6 g/(kg·d) of the prescription by gavage, while the other three groups received an equal volume of normal saline, twice daily for 15 consecutive days. The uterine appearance of pregnant mice was observed; individual embryo weight was recorded, and the embryo resorption rate was calculated. The proportion of activated CD41 + CD62p + platelets in platelet-rich plasma (PRP) was detected. Serum levels of thromboxane B2 (TXB2) and platelet factor 4 (PF4) were measured. Morphological changes of placental tissue were observed. The cell apoptosis rate of placental tissue was detected. The levels of malondialdehyde (MDA) content, tot al superoxide dismutase (T-SOD) activity, the interleukin-1β (IL-1β) and IL-18, the protein expressions of matrix metalloproteinase-3 (MMP-3), MMP-9, nuclear proliferation antigen-67 (Ki67), superoxide dismutase 2 (SOD2), Nod-like receptor protein 3 (NLRP3), apoptosis-associated speck-like protein containing a CARD (ASC), and caspase-1, as well as the mRNA expressions of superoxide dismutase 2(SOD2), NLRP3, ASC and caspase-1 of placental tissue were detected. RESULTS Compared with the normal control group and adjuvant control group, the model group showed significant decreases in individual embryo weight, T-SOD activity of placental tissue, and relative expression levels of SOD2 mRNA and protein ( P <0.05); while significant increases were observed in embryo resorption rate, proportion of activated CD41 + CD62p + platelets in PRP, serum levels of TXB2 and PF4, placental cell apoptosis rate, MDA content in placental tissue, relative protein expressions of MMP-3, MMP-9 and Ki67, relative protein and mRNA expression levels of NLRP3, ASC and caspase-1 as well as IL-1β and IL-18 levels ( P <0.05). The model group also exhibited irregular uterine morphology with localized atrophy in some uterine horns; placental tissue showed fragmentation of decidual cells, extensive vacuolization and necrosis of trophoblast cells in the labyrinthine zone, accompanied by vascular reduction. After intervention with Bushen huoxue formula, the above indicators were significantly reversed ( P <0.05), and uterine morphology and pathological damage were markedly improved. CONCLUSIONS Bushen huoxue formula can effectively improve pregnancy outcomes in aPL-RSA model mice, and its mechanism may be closely related to inhibiting platelet activation, reducing oxidative stress injury, enhancing trophoblast cell function, and suppressing the inflammatory response mediated by the NLRP3 inflammasome pathway.
2.Research progress of RNA m 6A modification in breast cancer
Junlong GUO ; Ruiqi ZOU ; Shaoqiang CHEN ; Yuxin LIANG ; Jing LI ; Sunan YONG ; Yuting HE ; Xiaobing XIE ; Ping LI
Journal of International Oncology 2025;52(8):532-537
Breast cancer is one of the most common malignant tumors among women worldwide, with an increasing incidence rate year by year, making it a significant public health concern. With the continuous advancement of tumor biology research, N 6-methyladenosine (m 6A) modification, as an important form of RNA modification, has attracted growing attention. The m 6A modification, the most prevalent RNA modification in eukaryotes, occurs in almost all types of RNA and plays a critical role in the occurrence, progression, and metastasis of breast cancer. It influences cell proliferation, apoptosis, and alterations in the tumor microenvironment, though the specific mechanisms underlying these effects require further in-depth investigation. Moreover, the specific patterns of m 6A modification demonstrate its potential as a novel biomarker for breast cancer, which could provide new directions for early diagnosis and prognosis evaluation.
3.Prognostic Value of Pan-immune Inflammatory Values in Patients with Ischemic Stroke Treated with Intravascular Intervention Combined with Tirofiban
Yuting CHEN ; Jun PU ; Bin YAN ; Renhua XIE
Journal of Kunming Medical University 2025;46(4):115-122
Objective To investigate the predictive value of pan-immune inflammatory values(PIV)in the prognosis of patients with ischemic stroke(AIS)treated with intravascular intervention combined with tirofiban.Methods 102 AIS cases admitted to the Department of Neurology of the First People's Hospital of Zhaotong from January 2019 to December 2023 and treated with intravascular intervention combined with Tirofiban were selected as the study objects.The patients were divided into the mild group(n=9),moderate group(n=45)and severe group(n=48)according to the score of NIHSS after the admission.According to the mRS 90 days after the treatment,the cases were divided into the good prognosis group(n=63)and poor prognosis group(n=38).Data were collected and analyzed.The predictive value of PIV level in patients with ischemic stroke treated with intravascular intervention combined with tirofiban was evaluated by ROC.The risk factors of adverse outcomes in patients with ischemic stroke treated with intravascular intervention combined with tirofiban were investigated by multivariate Logistic regression.Results A total of 102 patients with ischemic stroke were included.The PIV level in the good prognosis group 331.1(221.9,662.7)was lower than that in the poor prognosis group 1 025.7(451.5,1 841.3)(P<0.05).PIV level in the severe group was higher than that in the mild and moderate groups,and PIV level in the moderate group was higher than that in the mild group(P<0.05).The AUC(95%CI)of PIV for predicting adverse outcomes of intravascular intervention combined with tirofiban in patients with ischemic stroke was 0.750(95%CI 0.654~0.831),the cut-off point was 834.92,the specificity was 85.71%and the sensitivity was 65.79%.NIHSS score>18(OR=5.944 95%CI 2.011~19.137),PIV>834.92(OR=4.863,95%CI 1.410~18.685)were the risk factors for the poor outcome of intravascular intervention combined with tirofiban in patients with ischemic stroke(P<0.05).Conclusion Elevated PIV is closely related to the poor outcomes in patients with ischemic stroke treated with intravascular intervention combined with Tirofiban.High PIV and NIHSS scores are both risk factors for the poor prognosis in patients with ischemic stroke,and can be used as the sensitive indicators for the early identification.
4.Consensus on the use of DSA-guided intrathecal drug delivery system combined with Zi Wu Liu Zhu Acupoint Therapy for cancer pain management
Yi LU ; Cunzhi LIU ; Wujun GENG ; Xiaozhen ZHENG ; Jingdun XIE ; Guangfang ZHANG ; Chao LIU ; Yun LI ; Yan QU ; Lei CHEN ; Xizhao HUANG ; Hang TIAN ; Yuhui LI ; Hongxin LI ; Heying ZHONG ; Ronggui TAO ; Jie ZHONG ; Yue ZHUANG ; Junyang MA ; Yan HU ; Jian FANG ; Gaofeng ZHAO ; Jianbin XIAO ; Weifeng TU ; Jiaze SUN ; Yuting DUAN ; Bao WANG
Journal of Southern Medical University 2025;45(8):1800-1808
Objective To explore the efficacy of DSA-guided intrathecal drug delivery system combined with Zi Wu Liu Zhu Acupoint Therapy for management of cancer pain and provide reference for its standardized clinical application.Methods and Results Recommendations were formulated based on literature review and expert group discussion,and consensus was reached following expert consultation.The consensus recommendations are comprehensive,covering the entire treatment procedures from preoperative assessment and preparation,surgical operation process,postoperative management and traditional Chinese medicine treatment to individualized treatment planning.The study results showed that the treatment plans combining traditional Chinese with Western medicine effectively alleviated cancer pain,reduced the use of opioid drugs,and significantly improved the quality of life and enhanced immune function of the patients.Postoperative follow-up suggested good treatment tolerance among the patients without serious complications.Conclusion The formulated consensus is comprehensive and can provide reference for clinicians to use DSA-guided intrathecal drug delivery system combined with Zi Wu Liu Zhu Acupoint Therapy.The combined treatment has a high clinical value with a good safety profile for management of cancer pain.
5.Three-dimensional ultrasonography assessment of fetal auricle for predicting congenital aural atresia
Youlu LIU ; Ting LEI ; Yuting JIANG ; Ju ZHENG ; Qiao ZHENG ; Miao HE ; Lihe ZHANG ; Hongning XIE
Chinese Journal of Ultrasonography 2025;34(2):155-160
Objective:To explore the value of prenatal three-dimensional ultrasonography(3DUS)in displaying auricular morphotyping and dimensions for predicting congenital aural atresia(CAA).Methods:A retrospective collection of 227 fetuses who underwent ultrasound scans and retained auricular 3DUS volumes from January 2018 to December 2023 at the First Affiliated Hospital of Sun Yat-sen University was conducted. Fetuses were divided into two groups:a CAA group(52 fetuses,62 auricles)and a non-CAA group(175 fetuses,202 auricles),based on the presence or absence of external auditory canal identified through postnatal examination. According to 3DUS auricular contour and presence or absence of the concha,the auricles were divided into 4 types:type Ⅰ,C-shaped auricle with a concha;type Ⅱ,Irregular auricle with a concha;type Ⅲ,C-shaped auricle without a concha;type Ⅳ,Irregular auricle without a concha. And auricular length(AL)and width(AW)were measured to calculate the product of the auricular length and width(ALW). Normal reference ranges for ALW from the non-CAA group were developed. Differences of the auricular morphotyping and Z-score of ALW(ALWZ)were compared between the two groups. Receiver operating characteristic(ROC)curves were used to evaluate the diagnostic efficiency of auricular morphotyping,ALWZ and the regression model. A Logistic regression model for CAA based on auricular morphotyping and ALWZ were established.Results:The auricular morphotyping and ALWZ between the two groups were different statistically(both P<0.05). The AUC of the auricular morphotyping and ALWZ predicting CAA were 0.960(95% CI = 0.923 - 0.997)and 0.975(95% CI = 0.959 - 0.991)individually. Formula for CAA prediction model combining the two indicators(5.379 × morphotyping - 2.386 × ALWZ - Conclusions:The auricular morphotyping and dimensions can effectively predict CAA.
6.Establishment and validation of an artificial intelligence model for ultrasound image quality control in early pregnancy
Yuting JIANG ; Qiao ZHENG ; Caixin HUANG ; Ting LEI ; Hongning XIE
Chinese Journal of Ultrasonography 2025;34(7):563-570
Objective:To develop a deep learning-based artificial intelligence system for assessing image quality in early pregnancy ultrasound,and to evaluate its performance in anatomical structure identification and quality control.Methods:A retrospective study was conducted by collecting 17 910 static ultrasound images of 8 quality-control planes from fetuses at 11 to 13 +6 weeks of gestation who underwent routine first-trimester ultrasound examinations at the First Affiliated Hospital of Sun Yat-sen University from June 2018 to June 2024. The dataset was divided into a training set(12 536 images),a test set(3 582 images),and a validation set(1 792 images)in a 7∶2∶1 ratio to develop a prenatal-screening artificial intelligence system(PSAIS)and to evaluate its performance in the automatic recognition and quality control of standard planes during early pregnancy. The average precision and mean average precision(mAP)were used to measure the model's ability to recognize the anatomical structures on each plane. Intraclass correlation coefficient(ICC)and Kappa statistics were used to assess the consistency between PSAIS and expert-level sonographers in both plane image quality assessment and standardization. The efficiency of PSAIS was also compared to manual quality control. Results:In the test set,the mAP values for recognizing the anatomical structures of the 8 quality-control planes all exceeded 0.800. In the validation set,PSAIS demonstrated moderate to good agreement with two experts in image quality evaluation:the ICC ranged from 0.713 to 0.843 for one expert and 0.678 to 0.788 for the other,while the Kappa values ranged from 0.590 to 0.768 and 0.530 to 0.702,respectively. In terms of plane standardization scoring,PSAIS showed particularly high agreement with expert ratings on the transventricular view(compliance rate 94.6%,Kappa=0.860)and the four-chamber cardiac view with blood flow(compliance rate 94.1%,Kappa=0.778),with agreement above 70% for the remaining planes. Compared with manual quality-control,PSAIS significantly increased processing speed:the total processing time was only 413 seconds,markedly less than the 77 008 seconds and 94 918 seconds required for manual QC( P<0.001). Conclusions:The PSAIS system performs well in recognizing and controlling the quality of standard ultrasound planes in early pregnancy,demonstrating high consistency with expert evaluations and significantly improved processing efficiency. It has potential application value in enhancing the quality and efficiency of early pregnancy screening.
7.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.
8.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.
9.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.
10.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.

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