Study On The Effect And Mechanism Of Penthorum Chinense Pursh Compound In Treating Liver And Gallbladder Damp-heat In Rats

The high development of intensive farming and non-standard feeding management,such as poor control of environmental risk factors,the use of high-fat,high-protein or moldy feed,as well as the abuse of antibiotics and non-standard drug use,can all cause liver damage to animals.The risk is greatly increased,leading to animal liver cell damage and necrosis,liver tissue disease,liver function decline,and ultimately the death of the animal.Clinically,animals can see mucous membranes,yellow skin,vomiting,abdominal distension,ascites,short yellow urine,uncomfortab01 le stools,or constipation.According to traditional Chinese veterinary medicine,the disease is invaded by dampheat toxins,causing damp-heat to accumulate liver and gallbladder.The principle of treatment should be to protect the liver and relieve jaundice,clear away heat and detoxify.Penthorum chinense Pursh(Pc P)is a traditional Miao medicine in China,which has the functions of clearing away heat and detoxification,removing blood stasis and removing dampness,protecting the liver and strengthening the spleen.Preliminary studies have shown that the extract of Pc P has a good therapeutic effect and anti-oxidant and antiinflammatory effects on acute liver injury(AHI)caused by carbon tetrachloride(CCl4)in dogs and liver injury in chickens caused by aflatoxin B1(AFB1).Therefore,the selfmade Penthorum chinense Pursh compound(PCPC)establishes a rat liver and gallbladder damp-heat model and explores its therapeutic effects and mechanism of action in the treatment of liver and gallbladder damp-heat in rats,providing a theoretical basis for the clinical use of livestock and poultry.1.Explore the therapeutic effect of PCPC on liver and gallbladder damp-heat model rats.The rat liver and gallbladder damp-heat model was established by the method of "dimethylnitrosamine(DMN)+ high temperature and humidity environment";the experiment was divided into blank group,model group,Penthorum chinense Pursh compound high-dose group(PCPC-H),Penthorum chinense Pursh compound middledose group(PCPC-M),Penthorum chinense Pursh compound low-dose group(PCPC-L),and Yin Chen Hao Tang(YCHT)group were treated with drugs for liver and gallbladder damp-heat rats and observed.The results showed that the rats in the blank group had bright coat,good spirits,normal diet,healthy body,and normal body temperature,feces,urine,abdominal circumference and body color;rats in the model group had mental fatigue,rough coat,and diet Symptoms such as declining,visible mucous membrane and skin yellowing,short yellow urine,dry stool,fullness of the abdomen,etc.After 14 days of administration,the mental state,food intake,drinking water and body temperature of the rats basically returned to normal,and the color of visible mucosa and skin basically returned to normal.The effective rate of each drug group was 100%,the cure rate of PCPC-H,PCPC-M was 100%,and the cure rate of PCPC-L was 75%.2.To explore the effect of PCPC on serum liver function indexes and liver histopathology in rats with liver and gallbladder damp-heat.Detect serum alanine aminotransferase(ALT),aspartate aminotransferase(AST),alkaline phosphatase(ALP),total bilirubin(TBIL)levels,and hematoxylin and eosin(HE)And Masson staining to make rat liver tissue sections and observe.Compared with the blank group,the model group’s serum ALT,AST,ALP,TBIL increased,a large number of hepatocytes necrosis,liver cord arrangement disorder,liver lobule destruction,white necrosis,a large number of inflammatory cell infiltration,liver tissue collagen The deposition increases and the fiber separation is obvious.Compared with the model group,rats’ serum ALT,AST,ALP,TBIL decreased after PCPC treatment,liver tissue morphology was significantly improved,and the number of damaged liver cells,inflammatory infiltration,and fibrosis were significantly reduced.It shows that PCPC can effectively reduce liver damage in rats with liver and gallbladder damp-heat and reverse the progress of liver fibrosis.3.Explore the effect of PCPC on serum inflammatory factors and oxidative stress levels in rats with liver and gallbladder damp-heat.The rat serum interleukin-1β(IL-1β),interleukin-18(IL-18),interleukin-6(IL-6),superoxide dismutase(SOD),glutathione(GSH),malondialdehyde(MDA)levels.The results showed that compared with the blank group,the activities of SOD and GSH in the serum of liver and gallbladder dampheat model rats were significantly reduced,and the difference was extremely significant(P<0.01);IL-1β,IL-6,IL-18 and MDA levels increased,-1β,IL-6,IL-18 were significantly different(P<0.01),and MDA was significantly different(P<0.05);compared with the model group,the serum SOD and GSH activities of PCPC-H and YCHT rats increased.The content of MDA decreased,and the difference was extremely significant(P<0.01);the activities of SOD and GSH in serum of PCPC-M and PCPC-L rats increased,and the difference was significant(P<0.05).Serum IL-1β,IL-6 and IL-18 of rats in each dose group of PCPC significantly decreased(P<0.01).It shows that PCPC effectively reduces the inflammation level in rats with liver and gallbladder damp-heat,reduces oxidative stress,and improves the body’s antioxidant level.4.Explore the effects of PCPC on the activation of macrophages and hepatic stellate cells(HSC)in liver tissues of rats with liver and gallbladder damp-heat.Immunohistochemistry was used to detect the positive expression of CD68 in rat liver macrophages and α-SMA in hepatic stellate cells.The results showed that compared with the blank group,the positive expressions of CD68 and α-SMA in the liver of the liver and gallbladder damp-heat model group were significantly increased,and the liver sinusoids,fibrous septum,and portal area showed strong positive expressions.Compared with the model group,PCPC at each dose The expression of CD68 in the liver tissues of YCHT rats and YCHT rats was significantly lower;the expression of α-SMA was significantly weakened.It shows that PCPC can effectively reduce the positive expression of CD68 and α-SMA in liver tissue,inhibit macrophage activation and HSC activation,improve the liver microenvironment,and reduce the inflammatory response and fibrotic damage in rats.5.The effect of PCPC on NF-κB and NLRP3 inflammasomes in liver tissues of rats with liver and gallbladder damp-heat were explored.Western Blot method was used to detect the relative expression of liver phosphorylated nuclear factor NF-κB-p65 subunit(p-p65)and Caspase-1(Caspase-1)protein,and q RT-PCR method was used to detect liver nucleotide binding oligomerization structure Domain-like receptor protein 3(NLRP3),Caspase-1,apoptosis-related dot-like protein(ASC)m RNA transcripts.The results showed that compared with the blank group,the relative expression levels of phosphorylated nuclear factor p-p65 and Caspase-1 protein in the liver tissue of the model group were significantly increased,and the transcription levels of NLRP3,Caspase-1,and ASC m RNA were increased.Very significant(P<0.01).Compared with the model group,PCPC can effectively reduce the relative expression of p-p65,Caspase-1 protein and NLRP3,Caspase-1,ASC m RNA transcription in liver tissue.It shows that PCPC can effectively inhibit the activation of NF-κB signaling pathway and the activation of NLRP3 inflammasome,thereby reducing the inflammatory response and fibrotic damage in rats.In summary,PCPC has a good therapeutic effect on liver and gallbladder damp-heat rats.Its mechanism of action may be through reducing macrophage activation,inhibiting hepatic stellate cell activation,and regulating the expression of corresponding proteins in the NF-κB/NLRP3 signaling pathway to improve the liver microenvironment and increase the body’s antioxidant enzymes SOD and GSH.Active,inhibit the mass production of peroxidation product MDA,and reduce the content of inflammatory factors IL-1β,IL-6 and IL-18.