CN104887724A - Method for preparing extract of corchorus capsularis seeds and application of extract - Google Patents

Abstract

The invention relates to a preparation method and an application of an extract of jute seeds of the genus sylvestris. The seeds of the succulent jute are crushed, and 5 times the weight of petroleum ether is added to extract and degrease at room temperature; after the degreasing is completed, 5-10 times the weight is used The ethanol aqueous solution is carried out reflux extraction 1～3 times to the jute seeds of the genus genus, each time 1～3h, collects the obtained extract, which is the ethanol extract of the ramie jute seeds, and preserves at -20 ℃, the extraction Drugs are used to inhibit the proliferation of human bladder cancer cells. Compared with the prior art, the present invention is more effective in inhibiting the growth of human bladder cancer cell line T24, and has little effect on normal bladder epithelial cells, and is equally effective in the treatment of bladder cancer, the comprehensive utilization of jute germplasm resources in my country and the increase in the income of hemp farmers. is of great significance.

Description

The preparation method and application of the extract of jute seeds

technical field

The invention belongs to the technical field of medicines, and in particular relates to a preparation method and application of a jute seed extract.

Background technique

Bladder cancer is the most common malignant tumor of the urinary system, and its incidence rate ranks ninth among malignant tumors in the world and ranks first among malignant tumors of the urinary system in my country. Although 70-80% of primary bladder cancers are non-invasive (also known as superficial), they are prone to recurrence after surgery, with a recurrence rate as high as 70%, and about 15% of patients develop invasive bladder cancer. At present, regular intravesical infusion of mitomycin (MMC) and other chemotherapeutic drugs after transurethral resection of bladder tumor (TUR-BT) is mainly used clinically to prevent the recurrence and progression of non-invasive bladder cancer, but the effective rate is low (only 30-50%) and problems such as large toxic and side effects. Therefore, there is an urgent need to develop new anti-bladder cancer drugs with high efficiency and low toxicity to improve the chemotherapy effect of bladder cancer and reduce the side effects.

Paclitaxel, camptothecin, vincristine and other anticancer drugs extracted from plants have become the first choice for many cancer treatments due to their unique mechanism of action and remarkable anticancer efficacy, accounting for about 30% of the market share of anticancer drugs. Therefore, searching for new anticancer drugs or lead compounds from plant resources has become one of the hot spots in the field of cancer treatment research. Finding new anti-cancer drugs from poisonous plants is seen as a shortcut.

Corchorus capsularis (Corchorus capsularis) belongs to the bast fiber crops of the genus Jute of Tiliaceae. It is an annual herbaceous plant. Bitter hemp leaves, Niu Niu Ci, Sanzhu grass, basil and hemp bones, etc. Round fruit species jute is native to tropical Asia and is widely cultivated in the south of the Yangtze River in China. Its bast fiber has the characteristics of good hygroscopicity and fast water loss. It is mainly used for textile sacks and burlap. Jute seeds are poisonous, and they are used as a cure for diarrhea, dysentery, boils and swelling in the folk.

According to the literature search, so far there has been no report on the use of the extract of jute seeds of Cymbidium jute to inhibit the proliferation of bladder cancer cells and related diseases.

Contents of the invention

The object of the present invention is to provide a method for preparing and application of a jute seed extract with better curative effect and less influence on normal bladder epithelial cells in order to overcome the above-mentioned defects in the prior art.

The purpose of the present invention can be achieved through the following technical solutions:

The preparation method of the fruit seed jute extract, adopts the following steps:

(1) pulverizing the succulent jute seeds, adding 5 times the weight of sherwood oil to extract and degrease at room temperature;

(2) After the degreasing is completed, use 5-10 times the weight of ethanol aqueous solution to carry out reflux extraction for 1 to 3 times, each time for 1 to 3 hours, and collect the obtained extract, which is the succulent jute seed The ethanol extract was stored at -20°C.

The pulverized jute seeds were extracted and degreased in petroleum ether for 48 hours.

Use ethanol aqueous solution to airtightly soak the degreased jute seeds at room temperature for 2 hours, then heat in a water bath to reflux temperature for reflux, filter and concentrate with a rotary evaporator to obtain an extract.

The volume fraction of the ethanol aqueous solution is 60-90%.

The described jute seeds of the genus rotundum are wild or cultivated seeds of the genus rotundum jute.

The prepared jute seed extract of the fruit of the fruit can be applied to the inhibition of the proliferation of human bladder cancer cells, and the following steps are adopted:

(1) Cells in logarithmic growth phase: Human bladder cancer cell T24 was inoculated in a 96-well plate at a concentration of 10,000 cells per well;

(2) After sticking to the wall overnight, administer the ethanol extracts of jute seeds of the genus Cymbidium at a concentration of 0.1, 0.5, 1, 5 and 10ug/ml, and positive for mitomycin at 5, 10, 50 and 100ug/ml For the control, 6 replicate wells were set up for each treatment, and the experiment was repeated 3 times;

(3) After 24 hours of administration, the MTT method was used to detect the proliferation and growth rate of human bladder cancer T24 cells, and the IC50 value was calculated, and compared with the results of mitomycin, the effect of the extract of jute seeds on the human bladder cancer on human bladder cancer was obtained. Results of inhibition of T24 proliferation.

The human bladder cancer cells T24 were purchased from the Cell Bank of the Chinese Academy of Sciences.

Compared with the prior art, the present invention finds through the in vitro MTT anticancer activity evaluation that the growth inhibitory effect of the jute seed extract on human bladder cancer T24 cells is more efficient than that of the clinical first-line drug mitomycin, and it is more effective on normal bladder cancer. Epithelial cells are less affected. Therefore, the extract of the jute seeds of Cymbidium spp. can be used to prepare anti-bladder cancer drugs, and has good development and application prospects.

It is the first public disclosure of the use of the extract of jute seeds of Cymbidium in the preparation of anti-bladder cancer drugs, and the ethanol extract of jute seeds of Cymbidium is more effective in inhibiting the growth of bladder cancer cells than the current clinical first-line drug mitomycin, and Little effect on normal bladder epithelial cells. Therefore, the ethanol extract of jute seeds can be used to prepare high-efficiency and low-toxicity anti-bladder cancer drugs, which has good development and application prospects. Significance, in the preparation process, the use of petroleum ether to extract and degrease the growth inhibition rate has been further improved.

Description of drawings

Figure 1 is the IC50 value of T24 cells treated with the extract of jute seeds and mitomycin for 24 hours.

Fig. 2 is the inhibiting situation of T24 cell growth after 5ug/ml of the fruit extract of jute seed and mitomycin acted for 24 hours.

Detailed ways

The present invention will be described in detail below in conjunction with the accompanying drawings and specific embodiments.

Example 1

Preparation of ethanol extract of jute seeds

Take 100g of jute seeds, crush them, add 500ml of petroleum ether to degrease at room temperature for 48 hours, then use 1000ml of 70% ethanol aqueous solution, soak in airtight at room temperature for 2 hours, heat in a water bath to reflux, extract for 3 hours each time, and reflux in total Extract 2 times. The ethanol extracts collected twice were filtered and then concentrated with a rotary evaporator to obtain extracts. Collect the extract (that is, the ethanol extract of the jute seeds of the genus Cymbidium) and store it at -20°C for future use.

Example 2

Inhibitory effect of ethanol extracts of jute seeds from Cymbidium spp. on the proliferation of human bladder cancer cells

Cells in the logarithmic growth phase: Human bladder cancer cells T24 (purchased from the Cell Bank of the Chinese Academy of Sciences) were seeded in a 96-well plate at a concentration of 10,000 cells per well.

After overnight attachment, 0.1, 0.5, 1, 5 and 10 ug/ml ethanol extracts of jute seeds were administered, and 5, 10, 50 and 100 ug/ml mitomycin were used as positive controls. For each treatment, 6 replicate wells were set up, and the experiment was repeated 3 times.

After 24 hours of administration, the proliferation and growth rate of human bladder cancer T24 cells was detected by MTT method, and the IC50 value was calculated.

As shown in Figure 1, after acting on T24 cells for 24 hours, the IC50 (half-inhibition of growth) dose of the rufous jute seed extract was 2.44ug/ml, only 16.6% of mitomycin 14.70ug/ml, reaching a very significant difference , indicating that the extract of jute seeds of the fruit of the fruit is more effective in inhibiting the proliferation and growth of bladder cancer cell T24 than the current clinical first-line drug mitomycin, and has a good development and application prospect. As shown in Figure 2, after 24 hours of action, the growth inhibition rate of 5ug/ml jute seed extract on T24 cells was 63.36%, which was 2.08 times that of mitomycin 30.39%, reaching a very significant difference. The above results show that the anti-bladder cancer activity of the extract of jute seeds of Cymbidium spp. is more efficient than that of the current clinical first-line drug mitomycin, and can be used to prepare anti-bladder cancer drugs, which has a good development and application prospect.

Example 3

The preparation method of the fruit seed jute extract, adopts the following steps:

(1) Pulverize the wild round fruit type jute seeds, add sherwood oil 5 times of its weight and extract and degrease at room temperature for 48h;

(2) After the degreasing is completed, utilize 5 times of weight, and the volume fraction is 90% ethanol aqueous solution at room temperature and airtightly soaks the round fruit kind jute seed 2h after degreasing, then is heated to reflux temperature in water bath and carries out reflux extraction 1 time, and extraction time is 3h , collect the obtained extract, which is the ethanol extract of jute seeds, and store at -20°C.

Example 4

The preparation method of the fruit seed jute extract, adopts the following steps:

(1) Pulverize the cultivated round fruit kind jute seed, add the sherwood oil of its 5 times of weight and extract and degrease 48h under room temperature;

(2) After degreasing, utilize 10 times of weight, volume fraction is 60% ethanol aqueous solution airtightly soaks the round fruit kind jute seed 2h after degreasing at room temperature, then is heated to reflux temperature in water bath and carries out reflux extraction 1 time, and extraction time is 3h , collect the obtained extract, which is the ethanol extract of jute seeds, and store at -20°C.

Finally, it is necessary to explain here that: the above examples are only used to further describe the technical solutions of the present invention in detail, and cannot be interpreted as limiting the protection scope of the present invention. Non-essential improvements and adjustments all belong to the protection scope of the present invention.

Claims (8)

1. the preparation method of round fruit kind jute seed extract is characterized in that, adopts the following steps: (1) pulverizing the succulent jute seeds, adding 5 times the weight of sherwood oil to extract and degrease at room temperature; (2) After the degreasing is completed, use 5-10 times the weight of ethanol aqueous solution to carry out reflux extraction for 1 to 3 times, each time for 1 to 3 hours, and collect the obtained extract, which is the succulent jute seed The ethanol extract was stored at -20°C. 2. the preparation method of the syringa jute seed extract according to claim 1, is characterized in that, the syringa jute seed after pulverizing is extracted and degreased in sherwood oil 48h. 3. the preparation method of the genus jute seed extract according to claim 1, is characterized in that, utilizes ethanol aqueous solution at room temperature to airtightly soak the genus jute seed 2h after degreasing, then is heated to reflux temperature in a water bath and carries out reflux , filtered and concentrated with a rotary evaporator to obtain the extract. 4. according to the preparation method of claim 1 and 3 described C. rufica jute seed extract, it is characterized in that, the volume fraction of described ethanol aqueous solution is 60-90%. 5. the preparation method of the succulent jute seed extract according to claim 1, is characterized in that, the described succulent jute seed is wild or cultivated succulent jute seed. 6. the preparation method as claimed in claim 1 prepares the application that obtains the seed extract of the genus Jute, is characterized in that, the extract is applied to the inhibition of the proliferation of human bladder cancer cells. 7. the application of the fruit seed jute extract according to claim 6, is characterized in that, adopts the following steps: (1) Cells in logarithmic growth phase: Human bladder cancer cell T24 was inoculated in a 96-well plate at a concentration of 10,000 cells per well; (2) After sticking to the wall overnight, administer the ethanol extracts of jute seeds of the genus Cymbidium at a concentration of 0.1, 0.5, 1, 5 and 10ug/ml, and positive for mitomycin at 5, 10, 50 and 100ug/ml For the control, 6 replicate wells were set up for each treatment, and the experiment was repeated 3 times; (3) After 24 hours of administration, the MTT method was used to detect the proliferation and growth rate of human bladder cancer T24 cells, and the IC50 value was calculated, and compared with the results of mitomycin, the effect of the extract of jute seeds on the human bladder cancer on human bladder cancer was obtained. Results of inhibition of T24 proliferation. 8. The application of the extract of C. argutifolia jute seeds according to claim 7, characterized in that, the human bladder cancer cell T24 is purchased from the Cell Bank of Chinese Academy of Sciences.