1.Influencing factors for recompensation in patients with decompensated hepatitis C cirrhosis
Danqing XU ; Huan MU ; Yingyuan ZHANG ; Lixian CHANG ; Yuanzhen WANG ; Weikun LI ; Zhijian DONG ; Lihua ZHANG ; Yijing CHENG ; Li LIU
Journal of Clinical Hepatology 2025;41(2):269-276
ObjectiveTo investigate the influencing factors for recompensation in patients with decompensated hepatitis C cirrhosis, and to establish a predictive model. MethodsA total of 217 patients who were diagnosed with decompensated hepatitis C cirrhosis and were admitted to The Third People’s Hospital of Kunming l from January, 2019 to December, 2022 were enrolled, among whom 63 patients who were readmitted within at least 1 year and had no portal hypertension-related complications were enrolled as recompensation group, and 154 patients without recompensation were enrolled as control group. Related clinical data were collected, and univariate and multivariate analyses were performed for the factors that may affect the occurrence of recompensation. The independent-samples t test was used for comparison of normally distributed measurement data between two groups, and the Mann-Whitney U test was used for comparison of non-normally distributed measurement data between two groups; the chi-square test or the Fisher’s exact test was used for comparison of categorical data between two groups. A binary Logistic regression analysis was used to investigate the influencing factors for recompensation in patients with decompensated hepatitis C cirrhosis, and the receiver operating characteristic (ROC) curve was used to assess the predictive performance of the model. ResultsAmong the 217 patients with decompensated hepatitis C cirrhosis, 63 (29.03%) had recompensation. There were significant differences between the recompensation group and the control group in HIV history (χ2=4.566, P=0.034), history of partial splenic embolism (χ2=6.687, P=0.014), Child-Pugh classification (χ2=11.978, P=0.003), grade of ascites (χ2=14.229, P<0.001), albumin (t=4.063, P<0.001), prealbumin (Z=-3.077, P=0.002), high-density lipoprotein (t=2.854, P=0.011), high-sensitivity C-reactive protein (Z=-2.447, P=0.014), prothrombin time (Z=-2.441, P=0.015), carcinoembryonic antigen (Z=-2.113, P=0.035), alpha-fetoprotein (AFP) (Z=-2.063, P=0.039), CA125 (Z=-2.270, P=0.023), TT3 (Z=-3.304, P<0.001), TT4 (Z=-2.221, P=0.026), CD45+ (Z=-2.278, P=0.023), interleukin-5 (Z=-2.845, P=0.004), tumor necrosis factor-α (Z=-2.176, P=0.030), and portal vein width (Z=-5.283, P=0.005). The multivariate analysis showed that history of partial splenic embolism (odds ratio [OR]=3.064, P=0.049), HIV history (OR=0.195, P=0.027), a small amount of ascites (OR=3.390, P=0.017), AFP (OR=1.003, P=0.004), and portal vein width (OR=0.600, P<0.001) were independent influencing factors for the occurrence of recompensation in patients with decompensated hepatitis C cirrhosis. The ROC curve analysis showed that HIV history, grade of ascites, history of partial splenic embolism, AFP, portal vein width, and the combined predictive model of these indices had an area under the ROC curve of 0.556, 0.641, 0.560, 0.589, 0.745, and 0.817, respectively. ConclusionFor patients with decompensated hepatitis C cirrhosis, those with a history of partial splenic embolism, a small amount of ascites, and an increase in AFP level are more likely to experience recompensation, while those with a history of HIV and an increase in portal vein width are less likely to experience recompensation.
2.Effect of Dendrobium officinale Polysaccharides in Reversing Hepatic Fibrosis in Rats via Notch Signaling Pathway and Ultrasound Evaluation
Ying XU ; Jina LUO ; Liuqing YANG ; Qianqian TIAN ; Jiamao CHENG ; Haiyan CHEN
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(21):70-77
ObjectiveTo investigate the effect of Dendrobium officinale polysaccharides (DOP) in reversing carbon tetrachloride (CCl4)-induced hepatic fibrosis (HF) in rats via the Notch signaling pathway and evaluate the therapeutic effect of DOP by ultrasound elastography. MethodFifty-six male SD rats were randomized into normal, model, colchicine (1×10-4 g·kg-1), Fuzheng Huayu powder (0.45 g·kg-1), and low-, medium-, and high-dose (0.05, 0.1, 0.2 g·kg-1) DOP groups (n=8). The rats in the model group and each treatment group were injected subcutaneously with a mixture of CCl4-olive oil (2∶3) once every 3 days for 10 weeks. After 6 weeks of modelling, the rats were administrated with corresponding drugs once a day for 4 weeks. Hematoxylin-eosin (HE) staining and Masson staining were employed to observe the pathomorphological changes of the liver tissue. An automatic biochemical analyzer was used to measure the serum levels of alanine aminotransferase (ALT), aspartate transaminase (AST), alkaline phosphatase (ALP), and total bile acids (TBA). Enzyme-linked immunosorbent assay (ELISA) was employed to measure the serum levels of hyaluronic acid (HA), laminin (LN), type Ⅲ precollagen (PC-Ⅲ), and type Ⅳ collagen (Col-Ⅳ). The mRNA and protein levels of α-smooth muscle actin (α-SMA), Notch1, Jagged1, and Hes1 in the liver tissue were determined by Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) and Western blot, respectively. The Young's modulus (YM) of the rat liver was measured by acoustic radiation force impulse (ARFI) elastography before and after treatment. Then, the correlations of YM with the serum levels of HA, LN, PC-Ⅲ, and Col-Ⅳ and the protein levels of α-SMA and Notch1 signaling pathway-related factors in the liver tissue were analyzed. ResultCompared with the normal group, the model group showed disordered arrangement of liver cell cords, obvious infiltration of inflammatory cells, appearance of a large number of fat vacuoles, and fibrous proliferation, elevated levels of ALT, AST, TBA, ALP, HA, LN, PC-Ⅲ, and Col-Ⅳ in the serum, and up-regulated mRNA and protein levels of α-SMA, Notch1, Jagged1, and Hes1 in the liver tissue (P<0.01). Compared with the model group, drug interventions alleviated the pseudolobule formation and the collagen deposition in confluent areas. Except that the serum level of ALT in the low-dose DOP group had no significant changes, drug interventions, especially high-dose DOP, lowered the levels of ALT, AST, TBA, ALP, HA, LN, PC-Ⅲ, and Col-Ⅳ in the serum and down-regulated the mRNA and protein levels of α-SMA, Notch1, Jagged1, and Hes1 in the liver tissue (P<0.05, P<0.01). The results of ARFI and correlation analysis showed that the YM of the liver tissue was increased in the model group (P<0.01) compared with that in the normal group, Compared with the model group, drug interventions decreased YM (P<0.01). YM was positively correlated with the expression levels of HA, LN, PC-Ⅲ, α-SMA, Notch1, Jagged1, and Hes1s (r=0.754, 0.734, 0.801, 0.885, 0.896, 0.757, and 0.800, respectively, P<0.01), and it had a moderate correlation with Col-Ⅳ (r=0.688, P<0.01). ConclusionDOP can reverse HF by down-regulating the Notch1/Jagged1/Hes1 signaling pathway. YM can be used as an indicator in the assessment of the efficacy of DOP against HF.
3.Multicenter study on the efficacy and safety of 3 kinds of Cephalosporin for injection from centralized procurement or non-centralized procurement
Lu LIU ; Cangsang SONG ; Xingde LI ; Yuqin DENG ; Panpan MAO ; Hanshu ZHANG ; Yanzhen DONG ; Yaoyao CHENG
China Pharmacy 2024;35(7):825-830
OBJECTIVE To compare the efficacy and safety of Cefazolin sodium for injection, Cefuroxime sodium for injection, and Ceftazidime for injection from nationally organized centralized drug procurement (hereinafter referred to as “centralized procurement”) and non-centralized procurement in patients with bacterial infection. METHODS The case data of hospitalized patients who had used 3 kinds of Cephalosporins for injection from centralized procurement or non-centralized procurement in the treatment of bacterial infections were retrospectively collected from 19 medical institutions in Kunming from January 2020 to September 2022. After balancing the baseline differences between the groups with the propensity score matching method, the effectiveness and safety differences of 3 kinds of Cephalosporins for injection from centralized procurement or non- centralized procurement were compared respectively. RESULTS After balancing the baseline differences among the groups, 394 cases in each group of Cefazolin sodium for injection from centralized procurement or non-centralized procurement, 472 cases in each group of Cefuroxime sodium for injection from centralized procurement or non-centralized procurement, 504 cases in group of Ceftazidime for injection from centralized procurement and 590 cases in group of non-centralized procurement were included in the analysis. In terms of effectiveness, there were no significant differences in clinical response rate, 72 h response rate, bacterial clearance rate, and the recovery rate of body temperature, white blood cell count, neutrophil count, neutrophil percentage, C-reactive protein, procalcitonin recovery between the centralized procurement group and non-centralized procurement group of Cefazolin sodium for injection and Cefuroxime sodium for injection (P>0.05). The proportion of patients in centralized procurement group of Ceftazidime for injection with C-reactive protein restored to normal reference range was significantly higher than that in non-centralized procurement group (46.9% vs. 27.9%, P<0.05), but there were no statistically significant differences in other effectiveness indicators among groups (P>0.05). In terms of safety, there was no statistical difference in the incidence of adverse drug reactions between centralized procurement group and non-centralized procurement group of 3 kinds of Cephalosporins for injection (P>0.05); the incidence of platelet count reduction in centralized procurement group of Cefazolin sodium for injection was significantly higher than non-centralized procurement group (20.7% vs. 7.1%, P<0.05), the incidence of eosinophilia elevation in centralized procurement group of Ceftazidime for injection was significantly higher than non-centralized procurement group (5.3% vs. 1.9%, P<0.05). In addition, there was no statistically significant difference in the abnormal rates of other laboratory indicators among the three types of injection Cephalosporins (P> 0.05). CONCLUSIONS The efficacy of 3 kinds of Cephalosporin for injection from centralized procurement is not inferior to non- centralized procurement varieties, and the safety is equivalent to that of non-centralized procurement varieties.
4.Dendrobium Officinale Polysaccharide Alleviates Hepatic Fibrosis via Epithelial-Mesenchymal Transition
Liuqing YANG ; Qin FAN ; Yajie BAI ; Ying XU ; Jina LUO ; Jiamao CHENG ; Haiyan CHEN
Journal of Sun Yat-sen University(Medical Sciences) 2024;45(1):76-84
ObjectiveTo investigate the effect of Dendrobium officinale polysaccharide (DOP)on CCl4-induced hepatic fibrosis(HF)and its mechanism. MethodsA total of 56 male SD rats were randomly divided into seven groups: normal group(NG),model group(MG),colchicine group(CG, 0.1 mg/kg), Fuzheng Huayu group(FG, 0.45 g/kg),low-dose DOP group(LDG, 0.05 g/kg),middle-dose DOP group(MDG, 0.1 g/kg)and high-dose DOP group(HDG,0.2 g/kg),with 8 rats in each group. HF rat model was established by subcutaneous injection with 40% CCl4 olive oil mixture, every 3-day for 10 weeks. At the end of the sixth week, the drug groups were treated with colchicine, Fuzheng Huayu and DOP solution by gavage respectively, once a day for 4 weeks. NG and MG groups were similarly handled with an equal amount of 0.9 % normal saline. Liver histopathology was detected using hematoxylin-eosin (HE), Masson and Sirius red staining; blood biochemistry was tested for liver function and four indicators of HF; RT-qPCR and Western Blot were used to measure the expression of α-SMA, Col-I, E-cadherin, and ZEB1 genes and proteins in the liver tissues of rats, respectively. ResultsHE, Masson, and Sirius red staining showed that the liver tissue of MG rats had typical pathologic features of HF, and the degree of HF was alleviated in LDG, MDG, and HDG rats, respectively. Liver function test results showed that the serum AST, TBIL, and AKP levels were significantly lower in LDG, MDG, and HDG, compared with those of the MG (P < 0.05 or < 0.01). Meanwhile, ALT levels in serum deceased remarkably except in LDG (P < 0.05 or < 0.01). The four results of HF showed that the serum HA, LN, PC-Ⅲ, and COL-Ⅳ levels in LDG, MDG, and HDG rats were significantly decreased compared with those of the MG (P < 0.05 or < 0.01). The relative expressions of α-SMA, COL-I, and ZEB1 genes and proteins were significantly decreased in the liver tissues of LDG, MDG, and HDG (P < 0.05 or < 0.01), and the relative expression of E-cadherin gene and protein increased (P < 0.05 or < 0.01). In addition, the expressions of HA, α-SMA, COL-I, ZEB1 and E-cadherin were dependent on the dose of DOP. ConclusionDOP alleviated the degree of CCl4 induced HF in rats by inhibiting the epithelial-mesenchymal transition in liver tissue.
5.Effects of canagliflozin combined with amiloride or benzamil on bone metabolism in rats with nephrotic syndrome
Wei HUAN ; Xiao LI ; Wenjuan HONG ; Xiaofei LUO ; Xuefang WANG ; Qian LI ; Jiamao CHENG
Journal of Clinical Medicine in Practice 2024;28(2):78-83
Objective To investigate the effects of sodium-glucose cotransporter 2 inhibitor (SGLT2i)canagliflozin and epithelial sodium channel inhibitor amiloride or benzamil in combination on the bone metabolism in the rats with the nephrotic syndrome (NS) induced by doxorubicin. Methods In the 49 male SD rats selected, 7 were randomly selected as control group (NG), and 42 were built as adriamycin-induced nephropathy model by injecting adriamycin through the tail vein, and were randomly divided into model group (MG group), canagliflozin group (KG group), benzamil group (BH group), amiloride group (AL group), canagliflozin+benzamil group (KB group) and canagliflozin+amiloride group (KA group), with 7 cases in each group. Each medication group was given intragastric administration according to the body weight of rats regularly every day, NG group and MG group were given equal amount of normal saline, the course of treatment was 6 weeks. The 24-hour urinary protein (24 h-UTP) of each group was detected one day before treatment to verify the successful preparation of the model. At 6 weeks after treatment, the 24 h-UTP, urine sodium (UNa), urinary potassium (UK) levels in urine and the albumin (ALB), triglyceride (TG), cholesterol (TC), low density lipoprotein (LDL), serum calcium (SCa), sodium (SNa), potassium (SK), 25-hydroxyvitamin D (25-OH-D), alkaline phosphatase (ALP), parathyroid hormone (PTH), procollagen type Ⅰ N-terminal propeptide (PINP) and beta C-terminal cross-linked telopeptides of typeⅠ collagen (β-CTX) levels in serum were measured, respectively. Results After successful modeling, the levels of 24 h-UTP, TG, TC, LDL and SCa in the MG group were significantly increased, while the levels of ALB, 25-OH-D, ALP, PINP and PTH were significantly decreased (
6.Monitoring results of plague vector fleas in Yulong ,Yunnan Province from 2019 to 2021
Yuqiong LI ; Fahong WANG ; Zongti SHAO ; Ennian PU ; Jian WANG ; Meiqi LIU ; Pan CHENG ; Zhengxiang LIU
Shanghai Journal of Preventive Medicine 2023;35(9):879-884
ObjectiveTo analyze the community structure and dynamics of parasitic fleas on the body surface of host animals and nested fleas in different seasons in the natural foci of wild rat plague in Yulong County,Yunnan Province, to explore the relationship between seasonal fluctuation of fleas and the prevalence of plague among animals, so as to provide evidence for plague prevention and control in the natural foci. MethodsNanxi Village, Huangshan Town, the core area of plague epidemic in Yulong County, was selected as the monitoring sample area in December 2019 (winter), August 2020(summer), October 2020(autumn) and March 2021(spring). Host animals were captured by rattrap at night and rat nests were excavated for collecting parasitic fleas on host animals and rat nest fleas in different seasons. Excel 2010 and SPSS 26.0 software were used to analyze the data, and Chi square test was used to compare the rate. Community ecological indicators were used to analyze the community structure and species diversity of the host animals and their parasitic fleas. ResultsA total of 355 vector fleas were captured, belonging to 7 species of 5 genera in 2 families. 441 small animals were captured and 138 rat body fleas were detected with the flea infection rate of 14.51% and the flea index 0.31. 96 effective rat holes were excavated and 217 fleas were detected with the flea infection rate of 35.42% and the flea index 2.26. Among the four seasons, the flea infection rates of rat body and rat nests were higher in summer and winter, showing a significant difference in general (χ2=15.851, P<0.01; χ2=16.398, P<0.01). The dominant species of flea community were Ctenophthalmus quadratus, Stenischia humilis, Neopsylla specialis and Frontopsylla spadix, with a dominance index of 0.434, 0.254, 0.180 and 0.110, respectively. The diversity and evenness of rat body fleas showed a distribution characteristic of decreasing, increasing and then decreasing again with season changes, and both were the highest in spring, while the ecological dominance showed an opposite trend. The diversity and evenness of rat nest fleas showed a distribution characteristic of increasing first and then decreasing in summer, autumn and winter, with the highest in autumn, while the ecological dominance was diametrically opposite. ConclusionThe fleas community structure is relatively stable in Yulong County, but the number of species in the community is unevenly distributed by seasons, and the status of dominant species is prominent. Local authorities should carry out timely preventive deratization and depulization measures according to the results of daily monitoring, so as to effectively avoid the prevalence and spread of plague among animals in plague foci.
7.Periplaneta americana extract CⅡ-3 induces senescence of leukemia K562 cells via SIRT1/mTOR signaling pathway.
Si-Yue HE ; Cheng-Gui ZHANG ; Heng LIU ; Yue ZHOU ; Zi-Yun TANG ; Zi-Ying BI ; Lu TIAN ; Min-Rui LI
China Journal of Chinese Materia Medica 2023;48(11):3039-3045
This study aims to investigate the role of slient mating-type information regulation 2 homolog 1(SIRT1)/tuberous sclerosis complex 2(TSC2)/mammalian target of rapamycin(mTOR) signaling pathways in the Periplaneta americana extract CⅡ-3-induced senescence of human leukemia K562 cells. K562 cells were cultured in vitro and treated with 0(control), 5, 10, 20, 40, 80, and 160 μg·mL~(-1) of P. americana extract CⅡ-3. Cell counting kit-8(CCK-8) and flow cytometry were employed to examine the proliferation and cell cycle of the K562 cells. Senescence-associated β-galactosidase stain kit(SA-β-gal) was used to detect the positive rate of senescent cells. Mitochondrial membrane potential was detected by flow cytometry. The relative mRNA level of telomerase reverse transcriptase(TERT) was determined by fluorescence quantitative PCR. The mRNA and protein levels of SIRT1, TSC2, and mTOR were determined by fluorescence quantitative PCR and Western blot, respectively. The results showed that CⅡ-3 significantly inhibited the proliferation of K562 cells and the treatment with 80 μg·mL~(-1) CⅡ-3 for 72 h had the highest inhibition rate. Therefore, 80 μg·mL~(-1) CⅡ-3 treatment for 72 h was selected as the standard for subsequent experiments. Compared with the control group, CⅡ-3 increased the proportion of cells arrested in G_0/G_1 phase, decreased the proportion of cells in S phase, increased the positive rate of SA-β-Gal staining, elevated the mitochondrial membrane potential and down-regulated the mRNA expression of TERT. Furthermore, the mRNA expression of SIRT1 and TSC2 was down-regulated, while the mRNA expression of mTOR was up-regulated. The protein expression of SIRT1 and p-TSC2 was down-regulated, while the protein expression of p-mTOR was up-regulated. The results indicated that P. americana extract CⅡ-3 induced the senescence of K562 cells via the SIRT1/mTOR signaling pathway.
Humans
;
Animals
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Periplaneta
;
Sirtuin 1/genetics*
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K562 Cells
;
Signal Transduction
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TOR Serine-Threonine Kinases/genetics*
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RNA, Messenger
;
Mammals
8.Therapeutic Effect of Canagliflozin on Nephrotic Syndrome and Its Ultrasonic Evaluation
Wen-juan HONG ; Hong-jun LI ; Jiu-lin ZOU ; Wei HUAN ; Xiao LI ; Jia-mao CHENG ; Hai-yan CHEN
Journal of Sun Yat-sen University(Medical Sciences) 2023;44(1):71-77
ObjectiveTo investigate the therapeutic effect of antidiabetic drug canagliflozin (CGLZ) on adriamycin-induced nephrotic syndrome (NS) in rats, and the evaluation of contrast-enhanced ultrasound (CEUS) combined with color Doppler flow imaging (CDFI) during the treatment. MethodsA total of 56 male SD rats were randomly divided into normal group (NG), model group (MG), prednisone (PAT) group (PG), low-dose single CGLZ group (LSCG), high-dose single CGLZ group (HSCG), low-dose CGLZ + PAT group (LUCG) and high-dose CGLZ + PAT group (HUCG), with 8 rats in each group. The NS model in rats was induced by injecting adriamycin twice into the tail vein, and then the NS rats were treated by intragastric administration daily for 6 weeks with reference of PAT. Twenty-four hour urine total protein (24 h-UTP) was assessed one day before the start of oral administration and at the end of 2, 4 and 6 weeks after oral administration, respectively. CDFI and CEUS were performed on the right renal artery at the end of 6 weeks after oral administration, and the blood of abdominal aorta was taken for serological test the next day. ResultsCompared with those detection index of NG rats, the 24-hour UTP of MG rats increased (P<0.01), the serum ALB decreased and TG, TC, LDL increased (P<0.01), and CDFI shows that RRCT was thinner (P<0.01) and the renal artery blood flow indicators RA-PI, RA-RI, RA-S/D all increased (P<0.05), and CEUS image shows that the TIC curve parameters TTP, AT, AUC all increased and DPI decrease in MG rats (P<0.01). After drug treatment, compared with those detection index of MG rats, 24 h-UTP decrease in LSCG after 2 weeks (P<0.01), and decrease significantly in all drug groups after 6 weeks (P<0.01); the serological test results show that the serum ALB in all CGLZ groups increased (P<0.05), TG decrease in LSCG (P<0.01), TC and LDL also decrease in LUCG after 6 weeks (P<0.05); CDFI shows that the RRCT thinning degree in all CGLZ is reduced (P<0.01), and the RA-PI in LSCG, RA-RI in PG, and RA-S/D in PG, LSCG, HSCG and LUCG rats all decreased (P<0.05); CEUS shows that the TTP, AT and AUC of renal TIC curve in drug treatment groups all decreased (P<0.01), and the DPI in PG, HSCG, LUCG and HUCG rats increased (P<0.01). ConclusionsCGLZ has the effect of treating NS, and the small dose is the best. CEUS combined with CDFI can be used to evaluate the renal morphology and hemodynamic changes of NS model rats before and after drug treatment, which is helpful to guide clinical application.
9.Effects of EFHD2 protein deletion on tight junction proteins in mouse Sertoli cells
Xiaolin Ye ; Guolin Xu ; Tijun Qian ; Feng Qin ; Yuntao Wang ; Yuhang Cheng ; Wenzhen Zhao
Acta Universitatis Medicinalis Anhui 2022;57(11):1744-1749
Objective :
To study the effect of EFHD2 protein deletion in Sertoli cells on Occludin,a component of tight junction protein and the localization and expression of EF-hand domain family member D2 (EFHD2) in mouse testis.
Methods :
Total RNA and protein were extracted from adult mice's heart ,liver ,spleen ,lung ,kidney, brain and testis tissues.The mRNA and protein levels of EFHD2 in each organ tissue were detected by qRT-PCR and Western blot.Immunofluorescence and immunohistochemistry detected the localization and expression of EF- HD2 in testicular tissues.SiRNA interference was used to reduce EFHD2 in Sertoli cells to detect Occludin protein expression.
Results :
qRT-PCR showed that the expression of EFHD2 was the highest in the testis.Western blot results showed that the expression level increased in testis tissue.Indirect immunofluorescence and immunohisto- chemistry results showed that the protein was mainly distributed in Sertoli cells and co-localized with cytoskeletal Vimentin,indicating that the protein was expressed in Sertoli cells.After the decrease of EFHD2 protein expres- sion,Occludin protein expression also decreased.
Conclusion
The expression of EFHD2 protein in the testis is relatively high,mainly distributed in Sertoli cells of the testis,co-localized with Vimentin,and can affect the nor- mal expression of tight junction protein Occludin.It is suggested that EFHD2 can promote and maintain the junction structure of Sertoli cells and provide a stable microenvironment for spermatogenesis.
10.Anti-tumor Effect and Mechanism of Lycopus lucidus Polysaccharide on Non-small Cell Lung Cancer A549 Cells
Ting-ting CHENG ; Yan LI ; Gui-yuan CHEN
Chinese Journal of Experimental Traditional Medical Formulae 2022;28(3):83-90
ObjectiveTo study the anti-tumor activity and mechanism of Lycopus lucidus polysaccharide (LLP) in vitro. MethodCell counting kit-8 (CCK-8) assay was used to detect the inhibitory effect of LLP (0, 5, 10, 15, 20 g·L-1) on the proliferation of A549 cells at different time points (24,48,72 h). The migration and invasion abilities of A549 cells were detected by wound healing assay and transwell assay after LLP (10, 20 g·L-1) treatment for 24,48 h. Propidium iodide (PI) single staining was applied to determine the effect of LLP of different concentrations (10,20 g·L-1) on the cell cycle of A549. The apoptosis of A549 cells induced by LLP (10, 20 g·L-1) was detected by Annexin V-FITC/PI kit. Real-time fluorescence-based quantitative polymerase chain reaction (Real-time PCR) was adopted to measure effect of LLP (10, 20 g·L-1) on gene expression of cysteine aspartate protease-3 (Caspase-3),cysteine aspartate protease-8 (Caspase-8),cysteine aspartate protease-9 (Caspase-9),cyclin-dependent kinase-1 (CDK-1), and Cyclin B1 in A549 cells. Western blot was used to detect the effect of LLP on protein expression of Caspase-3,Caspase-8,Caspase-9,B-cell lymphoma 2 (Bcl-2)-associated X protein (Bax),CDK-1,cyclin-dependent kinase-4 (CDK-4),cyclin-dependent kinase-6 (CDK-6),Cyclin B1,and Cyclin D1 in A549 cells. ResultCompared with the blank group, the LLP group showed decreased proliferation, migration, and invasion of A549 cells (P<0.05, P<0.01), increased proportion of G0/G1 phase (P<0.05), enhanced apoptosis rate (P<0.05, P<0.01), elevated mRNA expression of Caspase-3,Caspase-8,and Caspase-9 (P<0.05,P<0.01), reduced mRNA expression of CDK-1 and Cyclin B1 (P<0.05,P<0.01), up-regulated protein expression of Caspase-3,Caspase-8,Caspase-9, and Bax (P<0.05, P<0.01), and down-regulated protein expression of Bcl-2, CDK-1, CDK-4, CDK-6, Cyclin B1, and Cyclin D1 (P<0.05, P<0.01). ConclusionLLP can inhibit the proliferation of A549 cells, block the cell cycle in the G0/G1 phase (also G2/M phase), and induce cell apoptosis via the mitochondrial apoptosis pathway and death receptor pathway.


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