The Effects Of The Herbs Of Supplementing Qi And Nourishing Yin Reducing Phlegm And Dredging Meridians On The Diabetic Fatty Liver's Rats

Objective: Diabetic fatty liver (DFL) refers to hepatic steatosis, which occurs based on the diabetes mellitus(DM), it belongs to nonalcoholic fatty liver disease(NAFLD)."the two hits"theory says: in the process of NAFLD, insulin resistance and lipid metabolism disturbance are the first hit, which made the liver sensitive to various damage factors; oxidative stress, lipid peroxidation and various kinds of damage factors are the second hit, which further caused the liver degeneration, inflammation, necrosis and fibrosis."The two hits"theory performed outstandingly in the course of DFL. The study made the DFL model by the way of high fat diet and middle doses streptozotocin(STZ) intraperitoneal injecting, being intervened by the herbs of supplementing qi and nourishing yin reducing phlegm and dredging meridians. Then observing the general conditions of the experimental animals, TG, TC and HDL in the serum, hepatic pathology form, the level of MDA and the activity of T-SOD in the hepatic tissue, the level of Adiponectin in serum, the level of Adiponectin and AdiR2 in hepatic tissue, to discuss the therapeutical effect of the herbs of supplementing qi and nourishing yin reducing phlegm and dredging meridians on the DFL and the possible mechanism, to provide the experimental basis for the disease of"emaciation-thirst", whose basic pathogenesis is qi-yin deficiency and collaterals abstraction by phlegm and blood stasis.Methods: 75 healthy male SD rats weighted 240~260g were divided into 5 groups: 12 in normal group, 18 in model group, 15 in low dose group, 15 in middle dose group and 15 in high dose group after being fed1 week normally. Except the normal group being fed on normal diet, the other groups were fed on high fat diet for 4 weeks. Then all rats except rats in normal group were single-intraperitoneal injected with STZ 35mg/kg, the rats in normal group were given the intraperitoneal injection with relevant dose of normal saline.Blood glucose levels were measured for three times continuosly(≥16.7mmol/L)on the third day after STZ injection to confirm the development of diabetes. The rats in low dose group, middle dose group and high dose group were respectively given by intragastric administration with the herbs of supplementing qi and nourishing yin reducing phlegm and dredging meridians 5.29g·kg-1·d-1, 10.58g·kg-1·d-1,21.16 g·kg-1·d-1 once per day for 8 weeks, the rats in normal group and model group were given by intragastric administration with the normal saline of relevant doses. Measuring the body weight and getting the blood of arteria femoralis without diet for 12 hours at the end of the 8th week after giving the medicine of the rats, getting the serum by separating the blood, measuring the TG, TC and HDL in serum; measuring the liver weight and calculating the liver index; taking the hepatic tissue and observing the pathological changes in different groups by the microscope; storing the hepatic tissue in -80℃fridge, using for detecting the level of MDA and the activity of T-SOD in hepatic tissue, detecting the level of Adiponectin in the serum by the way of ELISA, detecting the level of Adiponectin in hepatic tissue by the way of RT-PCR, detecting the level of AdiR2 in hepatic tissue by the way of Western blot.Results:1 General condition of experimental ratsDuring the experiment, 4 rats died in model group, 2 in high dose group, 1 in middle, 1 in low dose group and none in normal group separately. The death reason may be the ketoacidosis which caused by hyperglycemia and the disorder of viscera function. The rats in normal group whose weights grew progressively, ate normally, reacted sharply, looked brightly, hair glossy, and theirs dejecta were granular with the amount of urine normal. The rats in other groups had obvious phenomenons of polydipsia and polyuria, and their spirits were dropping gradually with reaction becoming slowly, eyes glazed, hair becoming matting, and dejecta becoming shapelessly sometimes.2 The body weight, liver weight and liver index(liver weight/body weight×100%)of the rats in different groupsThe results display: compared with normal group, the body weight and liver weight of the rats in model group, low dose group, middle dose group and high dose group obviously increased (P<0.05); compared with model group, the body weight and liver weight of the rats in every dose group obviously reduced (P<0.05); and there were no significant differences among the three different dose groups (P>0.05). compared with normal group, the liver index of the rats in model group, low dose group, middle dose group and high dose group obviously increased (P<0.05); compared with model group, the liver index of the rats in every dose group obviously reduced(P<0.05); and there were no significant differences among the three different dose groups(P>0.05). It prompts that the herbs of supplementing qi and nourishing in reducing phlegm and dredging meridians could obviously reduce the body weight, liver weight, liver index of the DFL rats and the fatty segeneration of the liver.3 The pathology changes of the rats'hepatic tissue in different groupsIn the normal group, the hepatic lobule were complete and well defined, the big and round nucleus and the equality cytoplasm could be seen in the hepatocyte, and the little lipid droplet could be seen occasionally in the hepatocyte which radially arranged around the central vein, the hepatic sinus could be seen clearly, the hepatic cord arranged orderly; in the model group, the boundary of the hepatic lobule were unclear, the fatty denaturation and swelling of the hepatocyte were serious, the hepatocyte turned bigger, the lipid droplet of different sizes made the nucleus aside in the hepatocyte, the interstices of the hepatic sinus were narrow, fibroplasticproliferation could be seen partially, which extended into the hepatic lobule, the hepatic cord arranged disorderly; in the groups of the herbs of supplementing qi and nourishing in reducing phlegm and dredging meridians, the structure of the hepatic lobule were clear, the hepatic sinusoid had little hypertrophy but the structure were normal, the fatty denaturation were moderate in the hepatocyte, compared with the model group, the amount of the lipid droplets in the hepatocyte obviously reduced, and the size of most lipid droplets was small, the hepatic cord arranged comparatively orderly. It prompts that the herbs of supplementing qi and nourishing in reducing phlegm and dredging meridians could improve the hepatic steatosis of the DFL rats.4 The levels of TG, TC and HDL in the serum of different groupsThe level of TG in the serum of different groups display: compared with normal group, the serum TG in model group, low dose group, middle dose group and high dose group obviously increased (P<0.05);compared with model group, the serum TG in every dose group obviously reduced(P<0.05); compared with high dose and low dose groups, the serum TG in every dose group obviously reduced(P<0.05), and there were no significant differences between high and low dose groups(P>0.05).The level of TC in the serum of different groups display: compared with normal group, the serum TC in model group, low dose group, middle dose group and high dose group obviously increased(P<0.05), compared with model group, the serum TC in every dose group obviously reduced(P<0.05), compared with high dose and low dose groups, the serum TC in every dose group obviously reduced(P<0.05), and there were no significant differences between high and low dose groups(P>0.05).The level of HDL in the serum of different groups display: compared with normal group, the serum HDL in model group, low dose group, middle dose group and high dose group obviously reduced(P<0.05), compared with model group, the serum HDL in every dose group obviously increased(P<0.05), compared with high dose and low dose group, the serum HDL in middle dose group obviously increased(P<0.05), and there were no significant differences between high and low dose groups(P>0.05).It prompts that the herbs of supplementing qi and nourishing yin reducing phlegm and dredging meridians could obviously reduced the TG and TC in the serum, increased the HDL in the serum of DFL rats. These may be one of the mechanisms of the herbs'therapeutical effection on the DFL.5 The level of MDA and the activity of T-SOD of the rats'hepatic tissue in different groupsThe level of MDA in the hepatic tissue of different groups display: the compared with normal group, the level of MDA in the hepatic tissue in model group, low dose group, middle dose group and high dose group obviously increased (P<0.05); compared with model group, the level of MDA in the hepatic tissue in every dose group obviously reduced(P<0.05); compared with low and middle dose group, the level of MDA in the hepatic tissue in high dose group obviously increased(P<0.05), and there were no significant differences between middle and low dose groups(P>0.05).The activity of T-SOD in the hepatic tissue of different groups display:compared with normal group, the activity of T-SOD in the hepatic tissue in model group, low dose group, middle dose group and high dose group obviously ruduced(P<0.05); compared with model group, the activity of T-SOD in the hepatic tissue in every dose group obviously increased(P<0.05); compared with low and middle dose group, the activity of T-SOD in the hepatic tissue in high dose group obviously reduced(P<0.05), and there were no significant differences between middle and low dose groups(P>0.05).It prompts that the herbs of supplementing qi and nourishing yin reducing phlegm and dredging meridians could obviously reduce the level of MDA and increase the activity of T-SOD in the hepatic tissue of DFL rats. These may be one of the mechanisms of the herbs'therapeutical effection on the DFL. 6 The level of in the serum, the level of Adiponectin and AdiR2 in hepatic tissue in different groupsThe level of Adiponectin in the serum of different groups display: compared with normal group, the serum Adiponectin in model group, low dose group, middle dose group and high dose group obviously reduced(P<0.05); compared with model group, the serum Adiponectin in every dose group obviously increased(P<0.05); compared with middle dose and low dose group, the serum Adiponectin in high group obviously reduced(P<0.05), and there were no significant differences between middle and low dose groups(P>0.05).The level of Adiponectin in the hepatic tissue of different groups display: the level of Adiponectin in the hepatic tissue had no significant differences among different groups.The level of AdiR2 in the hepatic tissue of different groups display:compared with normal group, the AdiR2 in the hepatic tissue in model group, low dose group, middle dose group and high dose group obviously reduced(P<0.05); compared with model group, the AdiR2 in the rats'hepatic tissue in every dose group obviously increased(P<0.05); compared with high dose and low dose group, the AdiR2 in the hepatic tissue in middle group obviously increased(P<0.05), and there were no significant differences between high and low dose groups(P>0.05).It prompts that the herbs of supplementing qi and nourishing yin reducing phlegm and dredging meridians could obviously increase the level of the serum Adiponectin and AdiR2 in the hepatic tissue of DFL rats, but it had no obvious effection on the level of Adiponectin in the hepatic tissue.Conclusions: 1 The herbs of supplementing qi and nourishing yin reducing phlegm and dredging meridians could obviously delay the fatty degeneration of DFL rats'liver and reduce the liver index. It had good therapeutical effects on DFL.2 The herbs of supplementing qi and nourishing yin reducing phlegm and dredging meridians could obviously reduce the level of serum TG, TC and increase the level of serum HDL of DFL rats.It prompts that the therapeutical effects of the herbs may be concerned with regulating plasma lipids.3 The herbs of supplementing qi and nourishing yin reducing phlegm and dredging meridians could increase the ability of antioxidation of the DFL rats, delay the second hit in the"two hits"theory-lipid peroxidation, and protect the liver from becoming degeneration, necrosis and fibrosis. This effect may be the mechanism of the herbs'protection of the liver.4 The herbs of supplementing qi and nourishing yin reducing phlegm and dredging meridians could obviously increase the level of the serum Adiponectin and the level of AdiR2 in the hepatic tissue of DFL rats, and this may be one of the mechanisms of the herbs'protection of the liver, but it had no obvious effection on the level of Adiponectin in the hepatic tissue.