1.Synbiotics adjust intestinalmicroecology to treat rat NASH and its effect on TLR4
Jieyi CAI ; Danping LUO ; Yang ZHANG ; Yu YUAN ; Xingxiang HE
Basic & Clinical Medicine 2017;37(9):1263-1269
Objective To observe the change of intestinal microflora on the process of nonalcoholic steatohepatitis(NASH),and to explore the synbiotics therapeutic effect on NASH.Methods Rats were administrated with high fat diet to establish NASH model.In the process of NASH rats modeling,the level of triglyceride (TG), total cholesterol (TC), high-density lipoprotein (HDL), low density lipoprotein (LDL), fasting blood sugar (FBS) and fasting insulin (FINS) was dynamically tested by automatic biochemical analyzer.The change of main intestinal flora was detected by 16 S rRNA fluorescence quantitative polymerase chain reaction.NAFLD activity score was calculated.HE staining was used to observe the hepaticpathological changes and the TLR4 expression was detected by using enzyme-linked immunosorbent assay and immunohistochemical method.Until the 4th,8th,10th weekin the process of NASH modeling, 10 rats were feeded with synbiotics for 2 weeks, and all of above indicators were tested and observed.Results 1)With the extension of a high-fat diet feeding time, the degree of hepatocyte steatosis obviously increased.NAFLD score was significantly heightened(P<0.01).2)Number of independent activities of rats significantly increased, the serological level of TG, TC, LDL, FBS and FINS were lower significantly after intervention with synbiotics for 2 weeks(P<0.05).3)Synbiotics intervention for two weeks significantly increased the amount of bifidobacterium and lactobacillus and decrease the amount of enterococcus significantly(P<0.05).4)The expression of TLR4 was gradually increased in the process of NASH rats modeling(P<0.05),but decreased after 2 weeks of the synbiotics-intervention (P<0.05).Conclusions Intestinal microecology change is closely related to the development of NASH,therefor, synbiotics could improve the quality of life and biochemical indicators of NASH rats through adjusting intestinal microecology and the expression level of TLR4 protein might been involved.
2.The correlation between chronic hepatic diseases and small intestinal inflammation
Lihao WU ; Meihui CHEN ; Jieyi CAI ; Yu YUAN ; Xingxiang HE
Chinese Journal of Digestive Endoscopy 2017;34(5):322-325
Objective To investigate the correlation between chronic hepatic diseases and small intestinal inflammation.Methods Patients who received capsule endoscopy in The First Affiliated Hospital of Guangdong Pharmaceutical University were divided into groups of liver cirrhosis,non-alcoholic fatty liver disease(NAFLD),chronic hepatitis and non-hepatic disease according to clinic data from August 2011 to August 2015.The severity of small intestinal mucosal inflammation was graded according to Lewis Scoring system and incidence of small intestinal lesions in different groups and Lewis scores were compared.The liver function was also graded with liver noninvasive scoring systems.Then the correlation between liver function damage and small intestinal lesions was investigated.Results A total of 338 cases were enrolled in the study,including 25 cases of liver cirrhosis,47 cases of NAFLD,20 cases of chronic hepaitis and 246 cases of non-hepatic disease.There were 22 (88.0%),36 (76.6%),12 (60.0%) and 78 (31.7%) cases with lesions in small intestine in the four group respectively with significant differences(P<0.001).Rate of small intestinal villi edema was significantly higher in liver cirrhosis group,NAFLD group,chronic hepatitis group than that in non-hepatic disease group(all P<0.017).Small intestinal villi edema was found mainly in the upper and one third of middle parts in small intestine (P =0.033).Lewis scores of liver cirrhosis group (190.80±228.42)and NAFLD group(125.38± 191.31) were higher than those of non-hepatic disease group (42.91±97.69,P=0.021,P =0.034).Forns score,FIB-4 score,NAFLD-FS score and Child-Pugh score were positively correlated with Lewis score (correlation coefficient:0.247,0.244,0.223,0.284respectively,all P<0.001).Conclusion Chronic hepatic diseases such as liver cirrhosis,NAFLD,chronic hepatitis might be risk factors for small intestinal mucosal inflammation,and the severity of chronic hepatic diseases may be positively correlated with that of small intestinal mucosal lesions.
3.The effect of siRNA targeting MIF on the growth of colorectal cancer xenografts and the life quality of tumor-bearing mice
Yamin WANG ; Lijing WANG ; Rongjiao YANG ; Jieyi CAI ; Lihao WU ; Xingxiang HE
Chinese Journal of General Surgery 2011;26(5):376-380
Objective To analyze the effect of siRNA targeting MIF( MIFsiRNA) on the growth of colorectal cancer xenografts and the life quality of tumor-bearing mice.Methods BALB/C mouse model carring colorectal cancer was established.Thirty mice were divided into three groups randomly and managed respectively with intratumor injection of DEPC water, MIFsiRNA(0.15 nmol/g) and non-specific siRNA (0.15 nmol/g), respectively twice a week for consecutively 4 weeks.Drinking water, fodder consumed and body weight was recorded daily, and tumor volume was measured once a week.Mice were sacrificed after four weeks.ELISA and immunohistochemistry were used to detect the expression of MIF in serum and in tumor tissues.Spectrophotometric detection was used to detect caspase-3 protein.TUNEL was used to detect apoptotic cells.Results MIF expression in serum in MIFsiRNA group was lower than the other two groups [(22 ± 6) ng/ml vs (32 ± 8) ng/ml and (33 ± 8) ng/ml, P < 0.01]; MIF expression in tissues was less than the other two groups [(85 ± 20) /500 vs.(423 ± 23) /500 and (442 ± 31) /500, P < 0.01]; Tumor was smaller than the other two groups at third and fourth week (P < 0.01) ; Tumor weight was significantly less than the other two groups [(1.93 ±0.21) g vs (4.40 ±0.30) g and (5.25 ±0.44) g, P<0.01]; Mice in MIFsiRNA group were healthier than the other two groups as judged by water and fodder consumption (P < 0.01 ) , while weight change was not significantly different among the three groups ( P > 0.05 ).Caspase-3 protein in tissues was higher than the other two groups [(0.74 ±0.06) μg vs (0.57 ±0.08) μg and (0.56 ±0.02) μg, P <0.01]; Apoptosis cells in tissues were higher than the other two groups [(12 ± 2)/ 100 个vs 0 and 0, P < 0.01].Conclusions Knockdowning MIF gene expression inhibits the growth of colorectal cancer xenografts and improves life quality of tumor-bearing mice, possibly by a mechanism in which MIFsiRNA activates caspase-3 promoting cell apoptosis.
4.Angiotensin-(1-7) protects H9c2 cardiac cells against high glucose-in- duced injury and inflammation by inhibiting the interaction between TLR4 activation and necroptosis
Weijie LIANG ; Meiji CHEN ; Jieyi HE ; Jianhao LI ; Jun CHEN ; Fei CHENG ; Jun LAN
Chinese Journal of Pathophysiology 2016;32(10):1750-1756
AIM:To investigate whether angiotensin-(1-7) [Ang-(1-7)] protects H9c2 cardiac cells against high glucose (HG)-induced injury and inflammation by inhibiting the interaction between Toll-like receptor 4 (TLR4) acti-vation and necroptosis .METHODS:The expression levels of receptor-interacting protein 3 ( RIP3;an indicator of necrop-tosis) and TLR4 were determined by Western blot .Cell viability was measured by CCK-8 assay.The activity of lactate de-hydrogenase ( LDH) in the culture medium was measured with a commercial kit .The releases of interleukin-1β( IL-1β) and tumor necrosis factor-α( TNF-α) were measured by ELISA .The intracellular level of reactive oxygen species ( ROS) was analyzed by 2 ’ , 7 ’-dichlorfluorescein-diacetate ( DCFH-DA ) stating followed by photofluorography .Mitochondrial membrane potential ( MMP) was examined by rhodamine 123 staining followed by photofluorography .RESULTS:After the H9c2 cardiac cells were treated with HG (35 mmol/L glucose) for 24 h, the expression of RIP3 was obviously increased . Co-treatment of the cells with 30μmol/L TAK-242 (an inhibitor of TLR4) attenuated the up-regulation of RIP3 induced by HG.Furthermore, the expression of TLR4 was significantly increased after the cells were exposed to HG for 24 h, and co-treatment of the cells with 100μmol/L necrostatin-1 ( Nec-1;a specific inhibitor of necroptosis ) and HG for 24 h attenua-ted the up-regulation of TLR4 expression induced by HG .Moreover, 1μmol/L Ang-(1-7) simultaneously blocked the up-regulation of the RIP3 and TLR4 induced by HG.On the other hand, co-treatment of the cells with 1μmol/L Ang-(1-7), 30 μmol/L TAK-242 or 100 μmol/L Nec-1 and HG for 24 h attenuated HG-induced injuries and inflammatory response , leading to the increase in the cell viability , and the decreases in the activity of LDH , ROS generation , MMP loss as well as the releases of IL-1βand TNF-α.CONCLUSION:Ang-(1-7) protects H9c2 cardiac cells against HG-induced injury and inflammation by inhibiting the interaction between TLR 4 activation and necroptosis .
5.Role of ATP-sensitive potassium channels-Akt pathway in hydrogen sulfide inhibiting high glucose-induced injury in H9 c2 cardiac cells
Weijie LIANG ; Jingfu CHEN ; Jieyi HE ; Mingcai SONG ; Shenglong YU ; Wenzhu ZHANG ; Dongdan ZHENG ; Xinxue LIAO
Chinese Pharmacological Bulletin 2016;32(4):530-536
Aim To investigate the role of ATP-sensi-tive potassium channels-Akt pathway in exogenous hy-drogen sulfide( H2 S) inhibiting the high glucose( HG)-induced injury in H9c2 cardiac cells. Methods The expression level of Akt protein was tested by Western blot assay. The cell viability was measured by cell counter kit-8(CCK-8 assay). The number of apoptotic cells was tested by Hoechst 33258 nuclear staining fol-lowed by photofluorography. The intracellular levels of reactive oxygen species ( ROS ) were detected by DCFH-DA staining followed by photofluorography. Mi-tochondrial membrane potential ( MMP ) was examined by JC-1 staining followed by photofluorography. Results H9c2 cells were treated with 35 mmol·L-1 glucose (high glucose, HG) for 0 ~24 h respectively. After treating for 3 h, the expression level of phosphorated ( p )-Akt protein began to be obviously reduced, the maximum reduced expression level was observed after the cells were exposed to HG for 24 h. Pretreatment of the cells with 50 μmol · L-1 pinacidil ( Pin, a KATP channel opener) or 400 μmol·L-1 NaHS( a donor of H2 S) prior to exposure to HG considerably blocked the down regulation of p-Akt expression level induced by HG. However, pretreatment with 1 mmol · L-1 KATP channel blocker glibenclamide( Gli) obviously attenua-ted the inhibitory effect of NaHS on HG-induced down-regulation of p-Akt expression level. On the other hand, the protective effects of NaHS against the HG-induced cardiomyocyte injury were markedly blocked by 30 μmol·L-1 Ly294002(an inhibitor of Akt), as indicated by the decrease in cell viability and MMP dissipation as well as the increases in the number of apoptotic cells and ROS generation. Conclution KATP channels-Akt pathway mediates the protective effect of H2 S against the HG-induced cardiac injury.
6.Interaction between necroptosis and p38MAPK pathway mediates high glucose-induced injury in H9 c2 cardiac cells
Weijie LIANG ; Jieyi HE ; Jun CHEN ; Shenglong YU ; Wenzhu ZHANG ; Mingcai SONG ; Jingfu CHEN ; Jianqiang FENG ; Xinxue LIAO
Chinese Pharmacological Bulletin 2016;32(8):1138-1143,1144
Aim To investigate the role of the interac-tion between necroptosis ( Nec ) and p38 mitogen-acti-vated protein kinase ( MAPK) pathway in the high glu-cose (HG)-induced H9c2 cardiac cells injury.Meth-ods The cell viability was measured by cell counter kit-8 assay .The intracellular level of reactive oxygen species ( ROS ) was tested by DCFH-DA stating fol-lowed by photofluorography .Mitochondrial membrane potential ( MMP) was detected by Rhodamine 123 stai-ning followed by photofluorography . The expression levels of receptor interaction protein 3 ( RIP3, an indi-cator of Nec ) and p38 MAPK protein were tested by Western blot assay .Results The treatment of H9c2 cardiac cells with 35 mmol? L-1 glucose ( high glu-cose, HG) for 24 h induced considerable injuries , in-cluding a decrease in cell viability , increases in ROS generation as well as MMP loss .The co-treatment of the cells with 100 μmol? L-1 necrostatin-1(Nec-1,a specific inhibitor of Nec ) and HG for 24 h or the pre-treatment of the cells with 3 μmol? L-1 SB 2 0 3 5 8 0 ( an inhibitor of p38MAPK) for 60 min before HG exposure attenuated the above injuries induced by HG .Moreo-ver, the treatment of the cells with HG for 1,3,6,9, 12 ,24 ,36 and 48 h significantly increased the expres-sion levels of RIP3, peaking at 24 h.The co-treatment of the cells with 100 μmol? L-1 Nec-1 or the pre-treatment of the cells with 3 μmol? L-1 SB203580 considerably blocked the up-regulation of RIP3 expres-sion induced by HG .On the other hand , the co-treat-ment of the cells with 100 μmol? L-1 Nec-1 alleviated the HG-induced up-regulation of the expression of p-p38MAPK.Conclusion The interaction between Nec and p38 MAPK pathway mediates the HG-induced inju-ry in H9c2 cardiac cells.
7.Hydrogen sulfide protects H9c2 cardiomyocytes against high glucose-in-duced injury by inhibiting necroptosis
Weijie LIANG ; Jieyi HE ; Wenzhu ZHANG ; Shenglong YU ; Jun CHEN ; Mingcai SONG ; Jingfu CHEN ; Dongdan ZHENG ; Xinxue LIAO
Chinese Journal of Pathophysiology 2016;32(3):385-391
AIM:To study whether hydrogen sulfide (H2S) protects H9c2 cardiomyocytes against high glucose ( HG)-induced injury by inhibiting necroptosis .METHODS:The protein levels of RIP3 ( an indicator of necroptosis ) and cleaved caspase-3 were determined by Western blot .The cell viability was measured by CCK-8 assay.The intracellular le-vels of reactive oxygen species (ROS) were detected by 2’, 7’-dichlorfluorescein diacetate staining followed by photofluo-rography.Mitochondrial membrane potential (MMP) was examined by rhodamine 123 staining followed by photofluorogra-phy.The number of apoptotic cells was observed by Hoechst 33258 nuclear staining followed by photofluorography .RE-SULTS:After the H9c2 cells were treated with HG (35 mmol/L glucose) for 0~24 h, the protein expression of RIP3 in the H9c2 cells was significantly increased at 3 h, 6 h, 9 h, 12 h and 24 h, reaching the maximum level at 24 h.Pretreat-ment of the cells with 400μmol/L NaHS (a donor of H2S) or co-treatment of the cells with necrostatin-1 (Nec-1;a speci-fic inhibitor of necroptosis) considerably blocked the up-regulation of RIP3 protein induced by HG.Moreover, pretreatment with NaHS or co-treatment with Nec-1 obviously inhibited HG-induced injuries , leading to an increase in the cell viability , and decreases in the generation of ROS and MMP loss .On the other hand , pretreatment with NaHS also reduced the num-ber of apoptotic cells and the protein level of cleaved caspase-3 in the HG-treated H9c2 cardiomyocytes .CONCLUSION:H2 S protects H9c2 cardiomyocytes against HG-induced injury by inhibiting necroptosis .
8.Opening of ATP-sensitive K+channels protects H9c2 cardiac cells against high glucose-induced injury and inflammation by inhibiting TLR4/NF-κB pathway
Weijie LIANG ; Meiji CHEN ; Jieyi HE ; Huimin HUANG ; Shenglong YU ; Jun CHEN ; Jingfu CHEN ; Mingcai SONG ; Xinxue LIAO
Chinese Journal of Pathophysiology 2016;32(7):1153-1160
AIM:To investigate whether the opening of ATP-sensitive K+(KATP) channels protects H9c2 car-diac cells against high glucose ( HG)-induced injury and inflammation by inhibiting the Toll-like receptor 4 ( TLR4 )/nu-clear factor-κB ( NF-κB) pathway.METHODS:The protein levels of TLR4 and NF-κB p65 were determined by Western blot.The levels of interleukin-1β(IL-1β) and tumor necrosis factor-α(TNF-α) were detected by ELISA.The cell viabil-ity was measured by CCK-8 assay.Mitochondrial membrane potential (MMP) was examined by rhodamine 123 (Rh 123) staining followed by photofluorography.The intracellular levels of reactive oxygen species ( ROS) were detected by 2′, 7′-
dichlorfluorescein-diacetate (DCFH-DA) staining followed by photofluorography.The number of apoptotic cells was ob-served by Hoechst 33258 nuclear staining followed by photofluorography.RESULTS: After the H9c2 cardiac cells were treated with HG (35 mmol/L glucose) for 24 h, the protein levels of TLR4 and phosphorylated NF-κB p65 ( p-NF-κB p65) were significantly increased.Pretreatment of the cells with 100 μmol/L diazoxide ( DZ, a KATP channel opener) for 30 min before exposure to HG considerably blocked the up-regulation of the TLR4 and p-NF-κB protein levels induced by HG.Moreover, co-treatment of the cells with 30 μmol/L TAK-242 (an inhibitor of TLR4) obviously inhibited the HG-in-duced up-regulation of the p-NF-κB p65 protein level.On the other hand, pretreatment of the cells with 100 μmol/L DZ had a clear myocardial protection effect, which attenuated the HG-induced cytotoxicity, inflammatory response, mitochon-drial damage, oxidative stress and apoptosis, evidenced by an increase in the cell viability, and decreases in the levels of IL-1βand TNF-α, MMP loss, ROS generation and the number of apoptotic cells.Similarly, co-treatment of H9c2 cardiac cells with 30μmol/L TAK-242 or 100μmol/L PDTC ( an inhibitor of NF-κB) and HG for 24 h also obviously reduced the above injuries and inflammation induced by HG.CONCLUSION: The opening of KATP channels protects H9c2 cardiac cells against HG-induced injury and inflammation by inhibiting the TLR4/NF-κB pathway.
9.Education via WeChat improves glucose control and pregnant outcomes in women with gestational diabetes mellitus
Ying SHAO ; Gengsheng HE ; Xianming XU ; Jieyi HE ; Peiying WU
Chinese Journal of Perinatal Medicine 2018;21(10):678-682
Objective To evaluate the effect of strengthening nutrition intervention in gravidas with gestational diabetes mellitus (GDM) with WeChat on blood glucose control and pregnant outcomes.Methods A total of 410 gravidas,diagnosed with GDM and treated in the Department of Clinical Nutrition of Shanghai General Hospital from October 2015 to April 2016,were enrolled and randomly divided into two groups (n=205).The control group received traditional nutrition clinic education only,while the intervention group was given strengthened nutrition education through WeChat in addition to traditional education.Blood glucose level and insulin dosage were followed up after one,two and four weeks of intervention.Pregnant outcomes and patient satisfaction were investigated on 42 d after delivery.T test,Chi-square test and non-parametric test were used for statistical analysis.Results (1) Two weeks after the intervention,the average 1-hour postprandial blood glucose in the intervention group was lower than in the control group [(7.46± 1.01) vs (7.68± 1.06) mmol/L,t=2.243,P=0.025].After 4 weeks,both 1-and 2-hour postprandial blood glucose levels of the intervention group were lower than those of the control group [(7.03±0.65) vs (7.33±0.63) mmol/L,t=4.629,P<0.05;(6.00±0.65) vs (6.21 ±0.62) mmol/L,t=3.153,P<0.05] and more gravidas achieved euglycemia [79.9% (151/189) vs 60.8% (113/186),x2=16.483,P<0.001].(2) Compared with the control group,the intervention group had a higher vaginal delivery rate [38.7% (72/186) vs 50.5% (95/188),x2=5.288,P=0.021] and a lower rate of postpartum complications [9.1% (17/186) vs 2.1% (4/188),x2=7.394,P=0.007].All of the gravidas in the intervention group were satisfied with the WeChat intervention except one lost to follow up [99.5% (203/204)].Conclusions Strengthening nutrition education through WeChat is much more effective than traditional nutritional outpatient education alone in order to achieve a better control of blood glucose and improve pregnant outcomes in GDM women.This intervention is highly acceptable to gravidas and can be further extensively applied in nutrition clinic.
10.Application of China consensus on the protocol of early gastric cancer screening in Guangdong province
Wenrui XIE ; Lihao WU ; Min ZHANG ; Yu CHEN ; Yu YUAN ; Jieyi CAI ; Xingxiang HE
Chinese Journal of Digestive Endoscopy 2020;37(7):491-494
Objective:To assess the application value of China consensus on the protocol of early gastric cancer screening in Guangdong province.Methods:A new quantitative scoring system was used in Cantonese residents who underwent early gastric cancer screening from March 2018 to March 2019. According to the scores of initial screening, patients were divided into high-risk, medium-risk and low-risk groups. The detection rates of early gastric cancer, precancerous diseases and precancerous lesions under gastroscopy in each group were compared. Chi-square test was performed for statistical analysis.Results:A total of 545 individuals were selected for gastroscopy, in which 32 cases were classified into high-risk group, 184 into medium-risk group and 329 into low-risk group. The results of gastroscopy examination showed that high-risk group had the highest detection rate of early gastric cancer (12.5%), followed by medium-risk group (1.1%) and low-risk group (0) ( χ2=41.85, P<0.01); the detection rates of precancerous diseases exhibited a similar pattern: high-risk group (60.9%) > medium-risk group (52.4%) > low-risk group (34.3%) ( χ2=18.00, P<0.01). The detection rates of precancerous lesions were 17.9%, 8.8% and 8.8%, respectively, with no significant difference ( χ2=2.58, P=0.28). In terms of the positive rate of endoscopy, high-risk group (71.9%) showed the highest positive rate, followed by medium-risk group (57.1%) and low-risk group (40.1%) ( χ2=21.54, P<0.01). Conclusion:China consensus on the protocol of early gastric cancer screeing is of application value for the screening of early gastric cancer and precancerous lesions in the populations at risk of gastric cancer in Guangdong province.