Cyclooxygenase inhibitory and antioxidant compounds from the mycelia of the edible mushroom Grifola frondosa.

Authors: Zhang Y, Mills GL, Nair MG.

Abstract: Bioactive Natural Products Laboratory, Department of Horticulture and National Food Safety and Toxicology Center, Biosystems Engineering, Michigan State University, East Lansing, Michigan 48824, USA.

The bioassay-guided isolation and purification of the hexane extract of the cultured mycelia of Grifola frondosa led to the characterization of a fatty acid fraction and three compounds, ergosterol (1), ergostra-4,6,8(14),22-tetraen-3-one (2), and 1-oleoyl-2-linoleoyl-3-palmitoylglycerol (3).

The composition of fatty acid fraction was confirmed as palmitic, oleic, and linoleic acids by GC-MS and by comparison with the retention values of authentic samples. The structures of compounds 1-3 were established by spectroscopic methods.

The fatty acid fraction and compounds 1-3 showed cyclooxygenase (COX) enzyme inhibitory and antioxidant activities. The inhibition of COX-1 enzyme by the fatty acid fraction and compounds 1-3 at 250 microg/mL were 98, 37, 55, and 67%, respectively.

Similarly, COX-2 enzyme activity was reduced by fatty acid fraction and compounds 1-3 at 250 microg/mL by 99, 37, 70, and 4%, respectively. The inhibitions of liposome peroxidation by the fatty acid fraction and compounds 1 and 2 at 100 microg/mL were 79, 48, and 42%, respectively.

This is the first report of compounds 2 and 3 from the cultured mycelia of G. frondosa.

The COX inhibitory activities of compounds 1-3 are reported here for the first time.

J Agric Food Chem. 2002 Dec 18;50(26):7581-5.

Adapted from the December 2005 edition of CancerWire:

Nutritional Supplement Reported to Help Cancer Patients Undergoing First Clinical Trial was the subject of the May 2005 edition of CancerWire. The referenced supplement, Poly-MVA, was developed by Dr. Merrill Garnett, who spent over 20 years in search of a non-toxic formulation that could be used to support more orthodox treatments. [The active complex in Poly-MVA, palladium lipoic acid complex, is frequently abbreviated LAPd; Lipoic Acid Palladium.]  Over the last several years a growing number of cancer patients, many with advanced, metastatic disease, have used it as a nutritional supplement. Many claim that they have derived a benefit.

James W. Forsythe M.D., H.M.D., a board certified oncologist and homeopathic physician, oversaw the first clinical study of LAPd as an adjuvant for Stage IV cancer patients. Dr. Forsythe’s clinical trial began enrolling patients in January, 2004 (23 months ago). This is an independent study which has an IRB (Investigational Review Board) approval pending in the state of Nevada. CancerWire spoke with Dr. Forsythe to obtain an update regarding his clinical results.

A total of 207 Stage IV cancer patients were enrolled in the study. Some chose to take the LAPd alone while others followed the recommended protocol for the study — taking the LAPd in addition to some form of conventional therapy (low dose chemotherapy).  It should be noted that all patients with metastatic (Stage IV) disease are in danger of dying at any time. In fact an appallingly small percentage of conventionally treated cancer patients survive longer than 18 months after the disease has metastasized.

72 of the 207 patients in Dr. Forsythe’s study unfortunately could not be saved, and 43 chose to discontinue within 1-2 months of enrollment.  Thus there are a total of 92 evaluable patients remaining in the study at the time of this report. While the reported response rates do not include the 43 patients who left the study, it is very important to note that Dr. Forsythe’s  reported response rate calculations have factored in the 72 patients who died.  *[See NOTE below]

There has been an overall response rate of 56% for patients who followed the recommended protocol and 50% for patients who chose to take LAPd alone. Among the 92 evaluable patients remaining in the study, none are currently experiencing a progression of disease. Every evaluable patient falls into one of the following three categories: Complete Remission (CR), Partial Remission (PR), Stable Disease (SD). The safety profile was reported as excellent. To say that these response rates compare favorably with response rates for Stage IV cancer patients who receive conventional therapy alone would be a gross understatement.

*NOTE:  When patients who are enrolled in a study do not survive (die during the study period), it is quite common for clinical investigators to remove those patients from their data and report on the surviving patients only — specfically, those patients who survive and continue to participate in the study. This rather deceptive practice of “fudging the numbers” is disingenuous at best and blatantly misleading at worst.  Investigators whose reported data include only those patients who survive are clearly attempting to make the response rates appear to be much better than they actually are.  The December 2005 edition of CancerWire stated that Dr. Forsythe’s reported response rates would be lower if they included the 72 patients who died. We are quite sure that this editorial error was an innocent oversight and that there was no deliberate intention on the part of CancerWire editors to mislead their readers. The fact is that Dr. Forsythe’s report does include those 72 patients.  Thus Dr. Forsythe’s reported response rates are entirely accurate – there has been no attempt to “fudge” the numbers in any way.

ScienceDaily (Apr. 14, 2008) — Both the reishi mushroom (Ganoderma lucidum; Lingzhi) and green tea have long held a place in traditional medicine in China and other Asian countries, for the general promotion of health and long life and for the treatment of specific diseases. More recent scientific studies have confirmed that both enhance the body¹s immune functions and hold the potential for treatment and prevention of many types of cancer.

Now a new study by Chinese scientists found that combining the active ingredients in the mushroom and the tea creates synergetic effects that inhibited the growth of tumors and delayed the time of death in mice with sarcomas.

Yan Zhang, of Pharmanex BJ Clinical Pharmacology Center in Beijing, reported the results of two studies at Experimental Biology 2008 in San Diego on April 8. The presentation was part of the scientific program of the American Society for Pharmacology and Experimental Therapeutics (ASPET).

Reishi grows in damp, sunless mountain areas and was once a rare commodity. Today Reishi, like green tea polyphenols, is manufactured as an extract. Zhang and her colleagues examined products sold as ReishiMax and Tegreen, made by Utah-based Nu Skin Enterprises. ReishiMax contains high concentrations of the active components in the mushroom itself and cracked spores of the mushroom, including polysaccharides (13.5 percent) and triterpenes (6 percent), and Tegreen is almost completely (98-99 percent) made of tea polyphenols.

In the first study, designed to look at cancer treatment, mice were first injected intraperitoneally with sarcoma cells and then were given either low, medium or high dosages of ReishiMax or low, medium, or high dosages of a combination of ReishiMax and Tegree. A control group received neither product. All mice died of sarcoma development after treatment for 28 days. But treatment with the combination of reishi and green tea extracts delayed the animals¹ death within the first 12 days after sarcoma injection, compared to the animals receiving only ReishiMax.

In the second study, designed to look at cancer prevention, groups of healthy mice were given either low, medium or high dosages of ReishiMax or low, medium, or high dosages of a combination of ReishiMax and Tegreen. A control group received neither product. After receiving the specified treatment for 14 days, mice were given a suspension of sarcoma cells subcutaneously, while the treatments were continued. On day 28, the sarcoma tumors under the skin were recovered from the mice and weighed. The tumor weight was reduced by 45 percent with the combination therapy, but in a much lesser degree with only the Reishi, compared to tumors in mice receiving no treatment, further confirming the synergy of the two together.

Senior author of the paper, Dr. Jia-Shi Zhu of Pharmanex research Institute in Provo, Utah, says these findings suggest the therapeutic values of the combined use of the substances in both cancer prevention and adjuvant treatment.

Other authors include Li Zhang and Ying Qi of Pharmanex Shanghai R and D Center and Ningzhi Tan and Ling Gao of the Pharmanex BJ Clinical Pharmacology Center, Beijing.

In the clinic it is not unusual for us to see patients with Breast Cancer who are undergoing chemotherapy treatment. The following is an acupuncture protocol we would use for these patients. It is a basic formula that we would adjust based on the individuals symptomology.

The protocol is based on a 21 day chemotherapy infusion cycle, with TX1 and TX2 given during the first week of treatment. The first phase treatment deals with the tremendous overload that the Kidneys and liver experience handling the cytotoxic agents along w/ the dead cell debris, thus the treatment approach is to rapidily detoxify.

TX3 is done during week two and TX4 is done during week three. The treatment approach is to rapidly nourish and support the major organs and the immune system, to prepare these systems for the next round of chemotherapy.

Channel                  TX!              TX2               TX3                    TX4

Heart                      7                  3                     8                       2

Kidney                    6                  10                   4                       11

Ren                         12                6                     17                     3

Liver                       13                3                     8                       10

Ear Pnts*                St/Lu           Sp/Ki                Li/shenmen       He/Ki

Stomach                  36               39                    30                       37

Lung                        4                 7                      1                         9

Large Intestine         15              10                     11                       4

Spleen                      7                11                     3                         12

*St = stomach, Lu = lungs, sp = spleen, Ki = kidney, Li = liver, He = heart,

Document title

Differential control of growth, cell cycle progression, and gene expression in human estrogen receptor positive MCF-7 breast cancer cells by extracts derived from polysaccharopeptide I’m-Yunity and Danshen and their combination

Author(s)

HSIEH Tze-Chen ⁽¹⁾ ; WU Joseph M. ⁽¹⁾ ;

Author(s) Affiliation(s)

⁽¹⁾ Department of Biochemistry and Molecular Biology, New York Medical College, Valhalla, NY 10595, ETATS-UNIS

Abstract

The use of herbs has been the mainstay of treatment for a variety of human illnesses and is an essential part of culturally-based healing traditions in many societies and countries. Also, herbs, including Chinese herbs, are being incorporated as remedies for disease management and treatment in Western countries. In Traditional Chinese Medicine (TCM), herbal prescriptions are most frequently given to patients as complex formulations containing multiple herbs. Notably and unwittingly, this approach amounts to the administration of several chemical entities at once; the underlying theory is that interactions among the chemicals present in different herbs in a formula exert synergistic pharmacodynamic actions and neutralize the adverse effects and toxicities of specific individual chemicals. The effectiveness and mechanisms of this approach have not been well investigated or understood. A primary interest of this laboratory is to obtain experimental evidence that supports the fundamental mechanistic theme for the combinatorial herbal strategy described above and its potential application in preventing and treating breast cancer (BCa). In this study, we investigated the effects of 70% ethanolic extracts prepared from medicinal mushroom extract denoted I’m-Yunity and Danshen (Salvia miltiorrhizae Binge), alone and in combination, using MCF-7 cells as an in vitro model of estrogen receptor positive (ER⁺), low invasive BCa. Combination of I’m-Yunity and Danshen (referred to as I’m-Yunity-Plus) suppressed clonogenicity to a comparable degree as Danshen alone; both being significantly more active than I’m-Yunity. However, extract of Danshen was more active in inhibiting MCF-7 cell growth than I’m-Yunity-Plus. In comparison, I’m-Yunity elicited less growth inhibition. Flow cytometric analysis showed that I’m-Yunity-Plus induced partial block of G₁/S transition in MCF-7 cells, whereas Danshen slowed down cell progression from G₁/S into G₂/M phases of the cell cycle. Treatment by I’m-Yunity did not affect cell cycle progression in MCF-7 cells; however, it promoted active induction of apoptosis. In addition, treatment with Danshen alone resulted in a pronounced reduction in the expression of Rb, cyclin Dl, and p53, and also led to a diminution of p65 and p50 forms of NF-κB. The pronounced suppressive effects of Danshen on expression of the aforementioned genes were largely attenuated in cells treated with I’m-Yunity-Plus suggesting that ingredients in Danshen must have interacted with those in I’m-Yunity as to culminate in neutralization of the gene suppressive effects of Danshen. Additional support for such interactions was obtained by targeted cDNA array analysis using human tumor metastasis and BCa/ER signaling gene arrays. Taken together, our results are consistent with the interpretation that interaction exists between Danshen and I’m-Yunity and that I’m-Yunity-Plus may have efficacy in the treatment of BCa, particularly for patients with ER⁺ status.

Journal Title

International journal of oncology   ISSN 1019-6439 

Source

2006, vol. 29, n^o5, pp. 1215-1222 [8 page(s) (article)] (37 ref.)

MATRINE AND OXYMATRINE

Subjects Of Chinese Research

by Subhuti Dharmananda, Ph.D., Director, Institute for Traditional Medicine, Portland, Oregon

Matrine and oxymatrine are the two major alkaloid components found in sophora roots. They are obtained primarily from Sophora japonica (kushen), but also from Sophora subprostrata (shandougen), and from the above ground portion of Sophora alopecuroides. The matrines were first isolated and identified in 1958; they are unique tetracyclo-quinolizindine alkaloids (see Figure 1) found only in Sophora species thus far. An intensive investigation into the pharmacology and clinical applications of these alkaloids has gone on for the past decade and remains one of the focal points of Chinese medical research. The main clinical applications are treatment of people with cancer, viral hepatitis, cardiac diseases (such as viral myocarditis), and skin diseases (such as psoriasis and eczema).

The crude herb and crude hot-water extracts of sophora have been available in the West for more than 25 years. An alkaloid fraction of sophora roots containing a standardized level of oxymatrine and matrine (20%) was first introduced by the Institute for Traditional Medicine, and made available to practitioners in tablet form under the name Oxymatrine (White Tiger) in 1998. It has been used without reported side effects. In China, the alkaloids are often given by injection, but this method of administration is not acceptable in the West, so oral dosing is used here instead. When taken orally, much of the oxymatrine is converted to matrine; to get high blood levels of oxymatrine, it must be given by injection. However, it is unclear whether oxymatrine is clinically more effective than matrine. Chinese researchers have also used the alkaloids in capsule form, with results that appear similar to the injection. Sophora is also administered in complex formulas made as decoctions and taken orally.

Sophora japonica contains about a dozen alkaloids, with matrine and oxymatrine being by far the highest, together comprising about 2% of the dried root stock (most of it in the form of oxymatrine), followed by closely related alkaloids: mainly sophocarpine, but also minute amounts of sophoranol, sophoramine, sophoridine, allomatrine, isomatrine, and others (see Figure 2). These alkaloids were first reported as constituents of kushen in a series of publications from 1958-1978.

An overview of recent research on the pharmacology and clinical applications of the sophora alkaloids is presented below. In general, the dosage of the sophora alkaloids administered clinically is in the range of 400-600 mg per day.

VIRAL HEPATITIS

As described by Chen Yanxi and his colleagues at the Shanghai Second Medical University (1):

In recent years, oxymatrine has been recommended for treating chronic hepatitis B and chronic hepatitis C and has been shown effective in clinical practice. It has been utilized for these applications broadly, but the factors affecting its efficacy have not yet been determined.

Chen and his group gave oxymatrine injection to patients with hepatitis B. He confirmed that the viral load declined by this treatment, suggesting that oxymatrine served to inhibit the viral replication, not just reduce liver damage, which is the primary and more limited effect of many herbs used for hepatitis. Antiviral activity, for hepatitis C virus, was confirmed by the same group in cell culture tests (2). Clinical effectiveness for patients with hepatitis C had been reported earlier, including reduction of viral load (3). Oxymatrine may reduce death of liver cells damaged by means other than by inhibiting viral activity, as indicated in a pharmacology study of liver protective effects in immune-based liver damage (4).

Kang Junjie and Kang Suqiong, at the Treatment Center for Hepatic Diseases of the Amoy Municipal Hospital, reported that oxymatrine injection did not cause side effects other than rare local reactions at the injection site (5). They used this injection along with oral administration of complex Chinese herb formulas designed to match symptom-sign complexes and claimed that the effects were comparable to those attained with interferon therapy, except that adverse reactions were avoided. In particular, they claimed that the use of oxymatrine and Chinese herb formulas inhibited liver fibrosis (for further information on Chinese herbs for this purpose, see: Treatment and prevention of liver fibrosis). The inhibition of fibrosis appears to be a separate for additional function of sophora alkaloids beyond inhibiting viral activity. In laboratory animal studies carried out by Chen Weizhong and his colleagues at the Changzheng Hospital in Shanghai, matrine was shown to reduce the formation of liver fibrosis that was caused by chemical damage to the liver (6).

Thus, in relation to viral hepatitis, the sophora alkaloids appear to inhibit the viral replication, reduce destruction of liver cells, and protect against fibrosis. It has also been suggested that the alkaloids promote the flow of bile.

CANCER

Sophora subprostrata has long been regarded an anticancer herb in China. According to cancer specialist Chang Minyi (7), “Sophora subprostrata works through stimulating the anticancer immune mechanism of the patient and reinforcing his resistance against the growth of the tumor.” In 1998, Xu Xiangru and Jiang Jikai, working at the Congqing University of Medical Sciences, published a review of anticancer activity of sophora alkaloids (8). They relayed pharmacology studies indicating the alkaloids could inhibit growth of tumor cells directly, and could also affect immune functions. In clinical work, they described the use of sophora alkaloids for treating the side effect of leukopenia caused by cancer chemotherapy or radiation therapy and for treating certain cancers, notably uterine cervical cancer and leukemia. The herb is also considered an important ingredient in treatment of esophageal and laryngeal cancer. In a recent pharmacology study, it was reported that matrine could help leukemia cells differentiate into mature and normal white blood cells (9). Nonetheless, sophora alkaloids should not be relied upon as a sole treatment for cancer, but as an adjunct therapy, as there is no proof that the herb or these compounds are curative.

CARDIAC DISEASES

Sophora and its alkaloids are commonly used in China for treatment of heart arrhythmias (10). A possible mechanism of action is to help block sodium and calcium channels, a mechanism relied on by several antiarrhythmic pharmaceuticals. In a review of sophora alkaloid effects on the heart, Li Yan and He Liren, at the Affiliated Yueyang Hospital of Shanghai University of TCM, reported that:

1. sophora total alkaloids or matrine could counteract arrhythmia induced by many causes;
2. the total alkaloids or oxymatrine could regulate heart contractility;
3. the total alkaloids could dilate the coronary artery, increase blood flow, and improve oxygen delivery to cardiac cells; and
4. sophora root could counteract the coxsackie virus that causes myocarditis.

Li and He also relayed a clinical report from the Third Clinical Medical College of Beijing Medical University, about treatment of 167 patients with fast arrhythmia. The patients received each day 3-10 sophora root tablets (extract of 2 grams crude herb/tablet). The results indicated positive effects on various kinds of arrhythmia, such as premature systole, paroxysmal ventricular tachycardia, atrial fibrillation, and sinus tachycardia; the efficacy for premature systole appeared to be the best. This Beijing study and others were described also by Niu Kuizhi in his review (12) of clinical applications of sophora (kushen).

SKIN DISEASES

Sophora is frequently used in treatment of skin diseases, applied topically and consumed orally. One of the primary uses for topical therapy is treatment of vaginitis, particularly that due to candida infection (13). Recently, a topical liniment was developed combining sophora’s matrine with the anti-inflammatory flavonoid baicalin from scute (huangqin) for treatment of eczema, neurodermatitis, and psoriasis (14). This treatment was reported to be highly effective, especially for eczema, though the number of cases was small, so that further research must be done. The use of sophora for psoriasis is a promising new area (15). Zhang Junling and his colleagues at the Department of Dermatology, Tianjin Changzheng Hospital, studied the mechanism by which sophora alkaloids reduce psoriasis patches (16). They found that the alkaloids could inhibit keratinocytes, the cells that reproduce continuously to produce the characteristic scales.

REFERENCES

1. Chen Yanxi, et al., Relationship between serum load of HBV-DNA and therapeutic effect of oxymatrine in patients with chronic hepatitis B, Chinese Journal of Integrated Traditional Chinese and Western Medicine 2002; 22 (5): 335-336.
2. Chen YX, et al., The inhibitory effect of oxymatrine on hepatitis C virus in vitro, Chinese Journal of Liver Diseases 2001; 9 (Supplement): 12-14.
3. Li Jiqiang, et al., A preliminary study on therapeutic effect of oxymatrine in treating patients with chronic hepatitis C, Chinese Journal of Integrated Traditional and Western Medicine, 1998; 18(4): 227-229.
4. Xiang X, et al., Effect of oxymatrine on murine fulminant hepatitis and hepatocye apoptosis, Chinese Journal of Medicine, 2002; 115(4); 593-596.
5. Kang Junjie and Kang Suqiong, 30 cases of chronic hepatitis B treated with oxymatrine injection combined with syndrome differentiation of Traditional Chinese Medicine, Journal of Traditional Chinese Medicine, 2002; 43(1): 53.
6. Chen Weizhong, et al., Effect of matrine on experiment rat liver fibrosis, Chin Journal of New Drugs, 2000; 19(5): 410-412.
7. Chang Minyi, Anticancer Medicinal Herbs, 1992 Hunan Science and Technology Publishing House, Changsha.
8. Xu Xiangru and Jiang Jikai, Recent progress in anticancer bioactivity study of Sophora flavescens and its alkaloids, Chinese Journal of Integrated Traditional Chinese and Western Medicine 1998; 4 (3): 235-239.
9. Zhu Ningxi, et al., Study on inducing and differentiating function and mechanism of matrine on leukemia cells, ACTA Traditional Chinese Medicine and Pharmacology (Shanghai), 2001; 15(1): 43-44.
10. Ding Guangsheng, Anti-arrhythmia agents in traditional Chinese medicines, Abstracts of Chinese Medicine 1987; 1(2): 287-308.
11. Li Yan and He Liren, Pharmacological study of Sophora alkaloid actions on the cardiovascular system, Chinese Traditional and Herbal Drugs, 2000; 31(3): 227-229.
12. Niu Kuizhi, Pharmacology and clinical application of sophora flavescentis, International Journal of Oriental Medicine 1997; 22(1): 75-81.
13. Li Xiuying, et al., Treatment of 50 patients with candida albicans vaginitis by cortex sophorae, Chinese Journal of Integrated Traditional Chinese and Western Medicine 2000; 6 (2): 146-147.
14. Ding Ting, et al., The preparing and clinical applications of Complex Matrine Liniment, ACTA Chinese Medicine and Pharmacology, 2002; 30(2): 47-48.
15. Zhang Yaolong, Clinical study on matrine for the treatment of psoriasis, Hebei Journal of Medical Science, 1996; 69 (2): 590-591.
16. Zhang Junling, et al., Study on Apoptosis induced by oxymatrine in cultured keratinocytes, Chinese Journal of Dermatology and Venereology, 2000; 14(6): 367-368.

August 2002

Chemotherapy and Moxibustion

Chemotherapy is generally considered indispensable in the treatment of cancer, but it often lowers the quantity of white blood cells drastically. Two reports study the effect of moxibustion in preventing or treating this type of leukopenia.

In one study, 114 patients had lung cancer, cancer of the esophagus, lymphoma, or other malignant tumors. They all received 1-3 courses of chemotherapy, and as a result, their white blood cells fell below 4000/mm3.

Moxa cones the size of a half a Chinese date were burned on slices of ginger (0.2-0.3 cm thick) on dazhui Du14, geshu UB17, pishu UB20, weishu UB21, and shenshu UB23. Three cones were used on each point to make redness and a tolerable burning pain. Blisters were avoided. Treatment was given daily for 9 days. Other drugs to raise the white cell count were not taken during this time. Blood work was analyzed every three days. Moxibustion was stopped if the white cell count rose over 4000/mm3.

Results: 44.7% reached 4000/mm3 in three days. 29.8% reached it in 6 days. 16.7% made the goal in 9 days. 8.8% failed to rise to 4000/mm3. (JTCM 1993;13(4):266-7)

The 111 subjects in another study had suffered cancer of the lungs, stomach, breast, or colon. They were prescribed chemotherapy which was expected to result in bone marrow suppression. These subjects were randomly divided into two groups. After beginning chemotherapy, members of the moxibustion group received moxibustion on zusanli ST36 on one side (the side alternated each treatment). Medium-sized cones were stuck on with petroleum jelly. The cones were burned until the patient felt a small burning pain at the site. The number of cones varied for each patient. They were treated once a day.

The comparison group received three days of injections of granulocyte colony stimulation factor (G-CSF) in order to stimulate the bone marrow to produce more white blood cells.

Both groups had their blood drawn daily to measure WBC, starting on day 5. To summarize the results, the injections were more effective in preventing bone marrow suppression during days 5 through 9. In days 11 through 13, there were no significant statistical differences between the two groups.

It seems that moxibustion is a little slower but is able to perform the same job as G-CSF. Since G-CSF can have many side-effects and is certainly more expensive than moxibustion, some may want to choose moxibustion as an alternative or may want to use it along with G-CSF. (Jilin Zhongyi Yao 2005;2:33-34)