Application of isoquinoline alkaloid to prevention or treatment of diabetes

[0069] Example 1

[0070] In order to study the effect of corkine on blood glucose of type I diabetes, this experiment adopted a classic type I diabetes model, using C57BL mice to randomly divide into 3 groups, normal control group (10), STZ group (20). In the STZ group, intraperitoneal injection of STZ (50mg/Kg) was continued for 5 days, and fasting blood glucose was measured two weeks later. The model was successful if the blood glucose was greater than or equal to 13.8mmol/L. Then the STZ group was randomly divided into STZ group and phellodendron treatment group, 10 rats in each group. The pakine treatment group was given oral gavage (15mg/kg) for 10 weeks, and the fasting blood glucose was measured after 4 hours of fasting. figure 1 As shown, there are significant statistical differences.

[0071] General situation of animals:

[0072] Histopathological examination of pancreatic islets: paraffin section, HE staining, observation of pathological changes

[0073] Table 1. The effect of corkine on blood glucose of type I diabetes modeled by STZ.

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[0075] Note: *P<0.05, **P<0.01, ***P<0.001, compared with STZ group.

[0076] The results show that corkine can effectively reduce blood sugar in type I diabetes caused by STZ

[0077] Table 2. The effect of corkine on the number of pancreatic islets in STZ modeled mice

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[0079] Note: *P<0.05, **P<0.01, ***P<0.001, compared with STZ group.

[0080] Nine fields of pancreatic pathological sections were randomly selected for statistics. Attached figure 1 The results showed that the pancreas atrophy and tissue structure necrosis after STZ modeling, and the morphology and structure of pancreatic islets recovered significantly after treatment with corkine. Note: The scale is 20μm, and the magnification is 400 times.

[0081] Example 2

[0082] In order to study the effect of phellodendine on type II diabetes, this experiment uses a classic type II diabetes model, using C57BL mice to randomly divide into 3 groups, normal control group (10), model group (10), phellodendrine treatment group (10), among which the model group and the phellodendron treatment group were given a high-fat diet (basic feed with 20% lard) for 8 weeks, and the phellodendrine treatment group was given a high-fat diet and oral gavage (50mg/kg) at the same time After the administration, the fasting overnight, fasting blood glucose, glucose tolerance and insulin tolerance were measured after the administration. The experimental data is analyzed by variance analysis, and the result is expressed as x±s. The results show that the treatment effect of phellodendron is obvious, with significant statistical differences.

[0083] Experimental results:

[0084] Table 3. The effect of corkine on fasting blood sugar of type II diabetes modeled with high fat

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[0087] Note: ***P<0.001, compared with the normal group; *P<0.05, ***P<0.001, compared with the high-fat group

[0088] Glucose tolerance test is an oral glucose load test to understand the body's ability to regulate blood sugar after taking glucose. Glucose tolerance test can detect abnormal glucose metabolism early. It is currently recognized as the gold standard for diagnosing diabetes. When blood sugar is elevated but has not yet reached the diagnostic criteria for diabetes, OGTT can be used for differential diagnosis in order to determine whether there is diabetes. Under normal circumstances, the body has a set of mechanisms to maintain blood sugar. Oral glucose will quickly return to normal after a short-term increase in blood glucose, that is, normal glucose tolerance. The area under the corresponding oral glucose tolerance curve is small. Diabetic patients have impaired glucose utilization. After the blood glucose rises rapidly, the blood glucose declines slowly, that is, the area under the line corresponding to impaired glucose tolerance is larger. In this experiment, glucose tolerance is impaired.

[0089] Such as figure 2 The glucose tolerance curves of the corkine treatment group and the normal group are under the curve of the model group, and the corresponding area under the line is also lower than that of the model group, and has statistical significance.

[0090] Glucose tolerance test method: mice are fasted overnight, and after oral blood glucose of 2g/kg in mice, at 0, 15, 30, 60, 90, 120 min, blood is collected from the tail tip to determine the corresponding blood glucose, and the corresponding area under the line is calculated using GraphPad Prism software (AUC)

[0091] Insulin tolerance test method: mice are fasted overnight, mice are injected with 0.75IU/kg of insulin into the abdominal cavity at 0, 30, 60, 90, 120, and 150 minutes respectively. Blood is collected from the tail tip to determine the corresponding blood glucose. GraphPad Prism software is used to calculate the corresponding Offline area (AUC)

[0092] The raw data of glucose tolerance are as follows:

[0093] Table 4. Raw data of glucose tolerance in normal group mice

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[0096] Table 5. The original number of glucose tolerance and the corresponding area under the line in the high-fat group

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[0098] Table 6. Phellodendrine treatment of high-fat diet mice glucose tolerance raw count and area under the line

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[0100] figure 2.Effect of phellodendron on oral glucose tolerance in type II diabetic mice

[0101] Note: control is a normal diet group, HFD is a high-fat diet group, and HFD+Phellodendrine group is a high-fat diet

[0102] Table 7. Analysis of the area under the oral glucose tolerance line of corkine

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[0104] Insulin tolerance test is an experiment that reflects the body's sensitivity to insulin. The main feature of type II diabetes is insulin resistance and reduced sensitivity to insulin, that is, blood sugar is higher than normal body during the same time of injection of the same insulin, such as image 3 As shown, the sensitivity to insulin decreased after the high-fat model was created. After the same amount of insulin was injected, the blood glucose level at each point was higher than that of the normal group, and the corresponding area under the line was also higher than that of the normal group. After treatment with corkine at various times The blood glucose value of the point is lower than the model group, and the corresponding area under the line is lower than the model group induced by high fat. The results show that corkine can effectively increase insulin sensitivity and improve diabetes.

[0105] Table 8. Raw data of glucose and insulin tolerance in normal mice

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[0108] Table 9. Raw data of glucose and insulin tolerance in high-fat group mice

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[0110] Table 10. Raw data of glucose and insulin tolerance in mice treated with phellodendrine

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[0112] image 3 The effect of phellodendron on insulin tolerance in type II diabetic mice

[0113] Table 11. Offline statistical analysis of corkine on glucose and insulin tolerance in type II diabetic mice

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[0116] Note: *P<0.05, **P<0.01, ***P<0.001, compared with high fat group

[0117] It can be seen that compared with the normal group, the fasting blood glucose value of the model group mice was significantly higher, indicating that the type II diabetes model was successfully modeled. At the same time, compared with the model group, phellodendron can significantly reduce fasting blood glucose, increase glucose tolerance, and enhance insulin sensitivity.

[0118] Example 3

[0119] The therapeutic effect of phellodendron on hyperlipidemia and non-alcoholic fatty liver,

[0120] 1. Laboratory animals and methods:

[0121] C57BL mice, SPF grade, male, body weight (20±2)g, were randomly divided into two groups, the first group was 12 mice, which was the normal group, and was given normal feed. The remaining mice (44 mice) were divided into the second group. High-fat feed (basic feed with 20% lard, 1.25% cholesterol, 0.5% sodium cholate) is given free access to food and water. After 8 weeks of continuous feeding, 4 mice were sacrificed from the normal control group and the fatty liver model group. The serum and liver biochemical indicators of the two groups of mice, as well as liver histopathological sections were compared to determine whether the non-alcoholic fatty liver model mice were Successfully established.

[0122] Thirty-two successfully modeled mice were randomly divided into 4 groups, namely the normal control group (8), the model group (8), the corkine treatment group (8), and the positive control group (8 lovastatin group) ), continuous intragastric administration for 8 weeks. During the administration period, except for the normal control group fed with standard feed, the other groups continued to be given a high-fat diet until the end of the experiment. The phellodendrine treatment group was given a high-fat diet while being given by oral gavage (50mg/kg), and the positive control group was given a high-fat diet while being given by oral gavage of lovastatin (60mg/kg))

[0123] 2. Observation indicators:

[0124] a General situation of animals

[0125] The experimental animals were kept in SPF animal room, 12h light, 12h night, free eating and drinking, during the experiment

[0126] The animal is in normal condition.

[0127] b Liver function related indicators: serum ALT, AST,

[0128] c Blood lipid related indicators: TC, TG, HDL, LDL

[0129] d Pathological examination: HE staining

[0130] 3. Experimental results:

[0131] The results show that: Phellodendron can significantly reduce total cholesterol (TC) and triglycerides (TG) in plasma, increase high-density lipoprotein (HDL), reduce low-density lipoprotein (LDL), and reduce plasma aspartame The results of acid aminotransferase (AST), alanine aminotransferase (ALT), and pathological sections showed that it effectively alleviated non-alcoholic fatty liver degeneration caused by high-fat diet and improved liver damage caused by steatosis.

[0132] After the administration, the mice and blood lipid levels were measured. The experimental data is analyzed by variance analysis, and the result is expressed as x+s.

[0133] The results showed that phellodendron has a significant effect on reducing high fat, with significant statistical differences.

[0134] Table 12. Changes of blood lipid indexes of mice in each group

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[0136] Table 13 Changes of related indexes of liver function of mice in each group

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