Study On The Mechanism Of Depression's Influence On The Occurrence And Development Of Primary Liver Cancer And The Mechanism Of Action Of Xingpi Jieyu Prescriptio

1.Evidence-based evaluation of the effects of depressive disorder and antidepressants use on the incidence and development of primary liver cancer Study one:Objective:The aim of this study was to assess the effect of depressive disorder on primary liver cancer incidence and mortality risk.Methods:We systematically searched several English and Chinese databases,including the Cochrane Library,MEDLINE,Embase,PsycINFO,Web of Science,CNKI,CQVIP database,Wanfang database,and SinoMed,and 3 clinical trial registration platforms through December 2022.Studies that met our inclusion and exclusion criteria were included,and the quality of studies was assessed using the Newcastle-Ottawa scale.A random-effects model was used to calculate the pooled effect estimates for depressive disorder and liver cancer incidence risk,and a fixed-effects model was used to calculate the pooled effect estimates for depressive disorder and liver cancer mortality risk.Results:We included 11 studies with follow-up periods ranging from 3 to 33 years and sample sizes ranging from 251 to hundreds of thousands of people.The pooled result of depressive disorder on the incidence risk of liver cancer showed that depressive disorder did not affect the liver cancer incidence(RR 1.44,95%CI 0.97-2.13).In subgroup analysis,the cohort study subgroup and the middle-aged subgroup showed that depressive disorder increased the incidence risk of liver cancer,and the RRs were 1.60(95%CI 1.05-2.44)and 1.60(95%CI 1.04-2.47),respectively.The pooled result of depressive disorder on the liver cancer mortality risk showed that depressive disorder increased the risk of liver cancer mortality,with a RR value of 1.33(95%CI 1.12-1.57).In subgroup analysis,the Europe-Oceania subgroup showed depressive disorder was associated with a higher risk of liver cancer mortality(RR 2.82,95%CI 2.11-3.76),and the same goes for studies with up to 10 years of follow-up subgroup(RR 2.37,95%CI 1.85-3.04).Conclusion:This study shows that depressive disorder is not associated with overall risk of liver cancer incidence,but is correlated with an increased risk of liver cancer incidence in cohort studies which are higher level of evidence.Depressive disorder increases mortality risk in patients with liver cancer.However,the results lack robustness and need to be confirmed by other powerful prospective studies.2.Study on the mechanism of depressive disorder on the incidence and development of primary liver cancer and the mechanism of Xingpi Jieyu decoctionObjective:The purpose of this study was to investigate the effects of depressive disorder onStudy two:Objective:This study aimed to assess whether antidepressants use increases liver cancer risk.Methods:We systematically searched several English and Chinese databases,including the Cochrane Library,MEDLINE,Embase,PsycINFO,Web of Science,CNKI,CQVIP database,Wanfang database,and SinoMed,and 3 clinical trial registration platforms through May 2022.Studies that met our inclusion and exclusion criteria were included,and the quality of studies was assessed using the Newcastle-Ottawa scale.A random-effects model was used to calculate the pooled effect estimates and 95%confidence intervals(CIs).Results:We included 11 studies with a total of 132,396 liver cancer cases.The meta-relative risk(RR)for liver cancer associated with antidepressants use was 0.72(95%CI 0.59-0.86).In subgroup analyses,only selective serotonin reuptake inhibitors were negatively correlated with risk of liver cancer(RR 0.64,95%CI 0.51-0.79);both dose subgroups ≤365cDDD(RR 0.77,95%CI 0.69-0.85)and>365cDDD(RR 0.57,95%CI 0.40-0.81)were associated with lower liver cancer risk;only in patients with chronic viral hepatitis,the use of antidepressants reduced liver cancer risk(RR 0.70,95%CI 0.54-0.90).Conclusions:The result of the current meta-analysis shows antidepressants use is correlated with decreased cancer liver risk.However,the observed association needs to be verified by more powerful evidence from prospective,methodologically rigorous studies.the incidence and development of primary liver cancer and the mechanism of Xingpi Jieyu decoction.Methods:Study three:Male Sprague-Dawley rats were randomized into control group(CON),chronic unpredictable mild stress group(CUMS),hepatocellular carcinoma group(HCC),composite model group(CUMS+HCC),escitalopram group(ESC),and Xingpi Jieyu decoction group(XPJYF).CUMS methods and N-nitrosodiethylamine were used on rats to induce depression and hepatocellular carcinoma composite model.Sucrose preference test(SPT)and open field test(OFT)were conducted at baseline,week 4,week 8,week 12,and week 16 after modeling.Brain samples were obtained after week 16.We observed the pathological changes of the hippocampus by hematoxylin-eosin(HE)staining and toluidine blue staining.Study four:The grouping and intervention of rats were the same as those in study three.The liver index and spleen index were assessed after rats were sacrificed at week 8 and week 16.The serum levels of aspartate aminotransferase(AST)and alanine aminotransferase(ALT)were detected using an automatic biochemical analyzer.The serum interleukin-1β(IL-1β),interleukin-6(IL-6),interleukin-10(IL-10),and tumor necrosis factor-α(TNF-α)levels were detected by enzyme-linked immunosorbent assay(ELISA).We observed the pathological changes of the liver tissues by HE staining.Study five:The grouping and intervention of rats were the same as those in study three.Fecal samples were collected after week 16 after.Fecal DNA was sequenced using 16S ribosomal DNA gene sequencing.Western blotting(WB)was used to detect the expression of ZO-1 and Occludin of colonic tissues.We observed the pathological changes of the colon tissues by HE staining and Alcian blue staining.Study six:The grouping and intervention of rats were the same as those in study three.Targeted metabolomic analysis was performed to detect the serum bile acids after sacrifice.WB was performed for the detection of farnesoid X receptor(FXR),small heterodimer partner 2(SHP-2),peroxisome proliferator-activated receptor α(PPARα),bile salt export pump(BSEP)and cholesterol 7α-hydroxylase(CYP7A1).Results:Study three:(1)Behavioral evaluation:The sucrose preference rates of CUMS+HCC group were lower than those of CON group at each time point after modeling.At week 8.week 12 and week 16,the sucrose preference rates in ESC group and XPJYF group were higher than those in CUMS+HCC group.Speaking of the number of rearing and the number of entries to the central zone at each time point after modeling,these numbers were decreased in CUMS+HCC group compared to CON group.ESC group and XPJYF group had more numbers of rearing than CUMS+HCC group,and the number of entries to the central zone was higher in ESC group and XPJYF group than in the CUMS+HCC group,except at week 12.(2)The pathological of hippocampus:HE staining showed the number,morphology and distribution of neurons in each subarea of hippocampus in CON group were not abnormal.In CUMS+HCC group,the arrangement of neurons was disordered,the number of neurons decreased,a large number of neuronal cell bodies deformed,the nuclear became pyknosis,and some pathological fiber tangles could be seen.Compared with CUMS+HCC group,ESC group and XPJYF group were improved to varying degrees.Nissl staining showed that there were abundant Nissl bodies in the cytoplasm of hippocampal neurons in CON group.In CUMS+HCC group,a large number of neurons had irregular cell bodies,pyknosis and had lost substantial Nissl bodies.Nissl bodies increased in ESC group and XPJYF group.Study four:(1)Mortality rate:Survival analysis showed the mortality was higher in CUMS+HCC group than in CON group,CUMS group or HCC group.The mortality of ESC group and XPJYF group was lower than that of CUMS+HCC group.(2)Body weight,liver index and spleen index:At each time point after molding,the body weight of CUMS+HCC group was significantly lower than that of CON group.At week 8,week 12 and week 16,the body weight was higher in ESC group and XPJYF group than in CUMS+HCC group.At week 8 and week 16,the liver index and spleen index of CUMS+HCC group were higher than those of CON group.The liver index and spleen index were lower in ESC group and XPJYF group than in CUMS+HCC group at week 16.Although these two indexes tended to decrease at week 8 in ESC group and XPJYF group compared with CUMS+HCC,there was no statistical difference.(3)Liver tissues pathological staining:The structure of hepatic lobule was clear and the hepatocytes were normal in CON group.In CUMS+HCC group,the structure of liver tissue was disordered.The cancer tissue was nodular,and the cancer cells in the nodules were arranged in a nest-like fashion.The cell shape was extremely irregular,the ratio of nucleus to cytoplasm was increased,and the nuclear atypia was significant.In ESC group,pseudolobules were formed,and vacuolar degeneration and necrosis of hepatocytes were observed.In XPJYF group,no obvious pseudolobular formation was observed in the low-power microscope,but fibrous septa surrounding hepatocytes,vacuolar degeneration and necrosis of hepatocytes were observed in the high-power microscope.(4)Liver function:At week 8 and week 16,compared with CON group,AST and ALT increased in CUMS+HCC group.Liver function indicators were decreased or had a tendency to decrease in ESC group and XPJYF group compared with CUMS+HCC group.(5)Cytokines:Compared with CON group,the levels of IL-1β,IL-6,IL10 and TNF-α were significantly increased in CUMS+HCC group.Compared with CUMS+HCC group,the serum cytokines in ESC group and XPJYF group were decreased.Study five:(1)Gut microbiota sequencing:The Chao-1 index and Shannon index of CUMS+HCC group were lower than those of CON group.Compared with CUMS+HCC group,the Chao1 index was higher in XPJYF group.The difference of gut microbiota between CUMS+HCC group and CON group was the largest in principal co-ordinates analysis and nonmetric multidimensional scaling,while the difference could be reduced after the intervention of XPJYF.The LEfSe analysis showed that Bacteroidetes,Bacteroidia and Bacteroidales were significantly enriched in CUMS+HCC group compared with CON group,while Firmicutes in CON group was significantly higher than that in CUMS+HCC group.Compared with CUMS+HCC group,the above-mentioned microorganisms were significantly reduced in XPJYF group,and Firmicutes,Bacilli,Lactobacillales,Lactobacillaceae and Lactobacillus were obviously enriched in XPJYF group.(2)Intestinal mucosal barrier:Compared with CON group,the expressions of ZO-1 and Occludin in CUMS+HCC group were down-regulated.The expression of ZO-1 and Occludin in XPJYF group was up-regulated compared with CUMS+HCC group.(3)Colon tissues pathological staining:HE staining showed CON group had flat mucosa,closely arranged epithelium and evenly distributed intestinal crypts,and abundant goblet cells were seen.In CUMS+HCC group,the mucosa became thinner,the epithelial cells fell off,the lamina propria was exposed,the crypts were disordered and atrophied,and the mucus in goblet cells decreased.In XPJYF group,the surface of mucosa was uneven and showed finger-like changes,the crypt opening widened,the crypts atrophied,some of crypts expanded,and the goblet cell mucus decreased.Alcian blue staining showed that goblet cell mucus was abundant in CON group,but decreased in CUMS+HCC group.The mucus was increased in XPJYF group compared with CUMS+HCC group.Study six:(1)Serum bile acids:The main bile acids in rat serum were taurocholic acid,tauro-β-muricholic acid,β-muricholic acid,cholic acid,tauro-α-muricholic acid,taurochenodeoxycholic acid,and glycocholic acid.Compared with CON group,the levels of all serum bile acid molecules in CUMS+HCC group showed an upward trend,although not every molecule was statistically different.Compared with CUMS+HCC group,the levels of all serum bile acid molecules in XPJYF group showed a downward trend.(2)Bile acids metabolic pathway:Compared with CON group,the expressions of FXR,SHP-2,PPARa and BSEP in CUMS+HCC group were down-regulated,while the expression of CYP7A1 was upregulated.Compared with CUMS+HCC group,the expressions of FXR,SHP-2,PPARa and BSEP were up-regulated and the expression of CYP7A1 was down-regulated in XPJYF group.Conclusions:1.The depression degree of CUMS+HCC group is the most serious,and XPJYF can partly alleviate its depression-like behavior,partly repair its hippocampal neuron damage.2.Depression promotes the occurrence and development of liver cancer,and XPJYF can delay the progression of liver cancer3.There are significant changes of gut microbiota and intestinal mucosal barrier damage in CUMS+HCC model,while XPJYF could reverse the structural changes of gut microbiota and reduce intestinal mucosal barrier damage.And its mechanism may be related to bile acids metabolism pathway.4.The possible mechanism of depression promoting the occurrence and development of HCC is that the FXR pathway which regulates bile acids metabolism is disordered,resulting in bile acids stasis.High concentration of bile acids,especially hydrophobic bile acids,have cytotoxic and cancer-promoting properties.XPJYF can regulate the expression level of different proteins in FXR pathway,reduce the degree of bile acids stasis,and then slow down the progression of liver cancer.