阅读说明：本技术 6-o-烟酰半枝莲碱g在制备治疗肝纤维化的药物方面的应用 (Application of 6-O-nicotinoyl barbasco lotus alkaloid G in preparation of medicine for treating hepatic fibrosis ) 是由 蔡凤梅 陈亮 于 2020-07-01 设计创作，主要内容包括：本发明公开了6-O-烟酰半枝莲碱G在制备治疗肝纤维化的药物方面的应用。本发明发现,6-O-烟酰半枝莲碱G一方面可以抑制肝纤维化过程中关键细胞肝星状细胞的增殖,另一方面可以通过抑制肝星状细胞中纤维化关键蛋白的表达进而抑制其活性。因此,6-O-烟酰半枝莲碱G具有抗肝纤维化的作用,可以开发成抗纤维化药物。(The invention discloses application of 6-O-nicotinoyl scutellarin G in preparing a medicament for treating hepatic fibrosis. The invention discovers that the 6-O-nicotinoyl scutellarin G can inhibit the proliferation of a key cell hepatic stellate cell in the hepatic fibrosis process on one hand, and can inhibit the activity of the key cell hepatic stellate cell by inhibiting the expression of a fibrosis key protein in the hepatic stellate cell on the other hand. Therefore, the 6-O-nicotinoyl scutellarin G has the effect of resisting hepatic fibrosis and can be developed into an anti-fibrosis medicament.)

1. An application of 6-O-nicotinoyl barbasco lotus alkaloid G in preparing medicine for treating hepatic fibrosis is disclosed.

2. A method for treating hepatic fibrosis comprises administering 6-O-nicotinoyl barbascone G to hepatic fibrosis patient at effective dose.

Technical Field

The invention belongs to the field of medicines, and relates to application of 6-O-nicotinoyl barbasco lotus alkaloid G in preparation of a medicine for treating hepatic fibrosis.

Background

The liver is the main metabolic organ in the body and has strong regenerative capacity. Liver can usually regenerate after liver injury occurs, but if there is long-term injury and chronic inflammatory stimulus, the regenerative capacity of the liver will be greatly impaired and fibrosis will occur. Hepatic Fibrosis (HF) is a key step in the progression of chronic liver disease to cirrhosis, a compensatory repair response in the body. Various causes such as alcoholic liver disease, non-alcoholic steatohepatitis, viral hepatitis, chronic cholestasis and autoimmune hepatitis can develop into fibrosis to progress to cirrhosis and liver cancer, and serious threat is brought to the life health of patients.

Hepatic Stellate Cells (HSC) are key cells participating in an HF process, and in the process of injury repair, the HSC is activated, secretes a large amount of ECM, matrix metalloproteinase, metalloproteinase tissue inhibitor and the like, and simultaneously interacts with hepatic cells, hepatic sinus endothelial cells, kupffer cells and the like to cause fiber deposition, liver tissue structure remodeling and disorder and accelerate the HF process.

Therefore, the proliferation or the vitality of hepatic stellate cells is inhibited, and HF can be cured and relieved to a certain extent. At present, no activity report of inhibiting the proliferation of hepatic stellate cells by 6-O-nicotinoyl scutellarin G exists.

Disclosure of Invention

The invention provides application of 6-O-nicotinoyl barbasco in preparing a medicament for treating hepatic fibrosis in order to overcome the defects of the prior art.

The technical scheme for realizing the purpose of the invention is as follows:

an application of 6-O-nicotinoyl barbasco lotus alkaloid G in preparing medicine for treating hepatic fibrosis is disclosed.

A method for treating hepatic fibrosis comprises administering 6-O-nicotinoyl barbascone G to hepatic fibrosis patient at effective dose.

The beneficial technical effects are as follows:

the invention discovers that the 6-O-nicotinoyl scutellarin G can inhibit the proliferation of a key cell hepatic stellate cell in the hepatic fibrosis process on one hand, and can inhibit the activity of the key cell hepatic stellate cell by inhibiting the expression of a fibrosis key protein in the hepatic stellate cell on the other hand. Therefore, the 6-O-nicotinoyl scutellarin G has the effect of resisting hepatic fibrosis and can be developed into an anti-fibrosis medicament.

Drawings

FIG. 1 is a graph showing the inhibition rate of 6-O-nicotinoyl barbasco G on HSC-T6 cell proliferation at various concentrations;

FIG. 2 shows the inhibitory effect of 6-O-nicotinoyl barbaloin G on the expression of the key proteins α -SMA and TGF- β 1 for fibrosis in HSC-T6 cells at different concentrations.

Detailed Description

The following examples illustrate the essence of the present invention, but should not be construed as limiting the scope of the present invention.

First, test materials

Rat hepatic stellate cell HSC-T6 was purchased from Shanghai Bioengineering, Inc.

Fetal Bovine Serum (FBS), DMEM medium was purchased from Gibco, usa.

Penicillin streptomycin was purchased from shanghai xianpeng biotechnology limited.

Trypsin shanghai shi front biotechnology limited.

6-O-nicotinoyl barbasco G (CAS: 1206805-30-2) was purchased from Chenguan Biotech, Inc., Baoji city, and its HPLC purity was 98%.

Second, test method

1. Cell culture

Rat hepatic stellate cell HSC-T6 was revived and cultured in DMEM containing 10% FBS and 1% streptomycin at 37 ℃ in 5% CO₂Culturing in a constant temperature incubator, digesting and subculturing when the cell density reaches about 90%.

2. MTT method for determining proliferation inhibition effect of medicine on HSC-T6 cells

Taking rat hepatic stellate cell HSC-T6 in logarithmic growth phase, digesting and re-suspending to prepare the HSC-T6 with the concentration of 5 × 10⁴The cell suspension is inoculated in a 96-well plate, each well is 100 mu L, after 24h, a drug group is replaced by a culture medium containing 6-O-nicotinoyl hemibarbaloin G with different concentrations (2, 5 and 10 mu M and DMSO-assisted dissolution), a control group uses a culture medium which does not normally contain the drug (containing an equal volume of solvent DMSO), the zero adjustment wells are culture solution without cells, each group is provided with 3 multiple wells, after 48h of culture, 10 mu L of MTT solution (5mg/mL) is added into each well, incubation is carried out for 4h, supernatant is discarded, 150 mu L of DMSO is added into each well, oscillation is carried out for 5min, an absorbance value (A) is measured by a microplate reader at 570nm, zero adjustment is carried out by the zero adjustment wells, the proliferation inhibition rate of the different concentrations of 6-O-nicotinoyl hemibarbaloin G on HSC-T6 cells is calculated, and the proliferation inhibition rate is × 100 percent (1-A drug group/A control group).

3. Western blot method for determining inhibition effect of drug on expression of fibrosis key protein in HSC-T6 cells

Taking rat hepatic stellate cell HSC-T6 in logarithmic growth phase, digesting and re-suspending to prepare the HSC-T6 with the concentration of 1 × 10⁵The cells/mL cell suspension is inoculated in a 24-well plate, each well is 1mL, after 24 hours, the drug group is replaced by a culture medium containing 6-O-nicotinoyl barbaloidine G with different concentrations (2, 5 and 10 mu M, DMSO cosolvent), the control group is a culture medium containing no drug (containing the same volume of solvent DMSO) normally, and each group containsSetting 3 multiple wells, continuously culturing for 48h, collecting cells, washing with PBS, lysing the cells, collecting cell proteins, taking 50 μ g of the proteins, determining the expression levels of fibrosis key proteins α -SMA and TGF- β 1 in HSC-T6 cells of a comparative drug group and a control group by using a Western blot method, taking an internal reference of β -Actin, and taking pictures and analyzing by software.

4. Data processing

Statistical analysis was performed using SPSS17.0, data are presented as mean ± standard deviation, homogeneity of variance was tested using One-way ANOVA with One-way analysis of variance, and analyzed using Student's-t test, with p <0.05 representing significant differences.

Third, test results

1. Proliferation inhibition effect of HSC-T6 cell by medicine

The proliferation inhibition effect of 6-O-nicotinoyl hemibarbaloidine G on HSC-T6 cells at different concentrations is shown in Table 1 and figure 1, 2, 5 and 10 mu M6-O-nicotinoyl hemibarbaloidine G can inhibit the proliferation of HSC-T6 cells, and the inhibition effect is more obvious when the concentration of the drug is higher. The results show that 6-O-nicotinoyl scutellarin G can inhibit the proliferation of HSC-T6 cells.

TABLE 1 inhibition of HSC-T6 cell proliferation by 6-O-nicotinoyl barbasco liensine G at different concentrations

2. Inhibition of fibrotic key protein expression in HSC-T6 cells by drugs

The inhibition effect of 6-O-nicotinoyl hemibarbaloin G on the expression of key fibrosis proteins in HSC-T6 cells at different concentrations is shown in figure 2, 5 and 10 mu M6-O-nicotinoyl hemibarbaloin G can inhibit the expression of key fibrosis proteins alpha-SMA and TGF-beta 1 in HSC-T6 cells, and the inhibition effect is more obvious when the concentration of the drug is higher. The results show that 6-O-nicotinoyl scutellarin G can inhibit the expression of key fibrosis protein in HSC-T6 cells.